0% found this document useful (0 votes)

20 views76 pages

BB Notes

The document provides an overview of blood banking and transfusion medicine, detailing historical milestones, processes, and regulations governing blood collection, preservation, and transfusion. It outlines the roles of various blood components, their storage requirements, and the evolution of blood transfusion practices from ancient times to modern techniques. Additionally, it discusses the biological aspects of red blood cells, including their metabolism, membrane structure, and the importance of preservation methods to maintain viability during storage.

Uploaded by

Miggy Villaroman Acosta

We take content rights seriously. If you suspect this is your content, claim it here.

Available Formats

Download as PDF, TXT or read online on Scribd

0% found this document useful (0 votes)

20 views76 pages

BB Notes

Uploaded by

Miggy Villaroman Acosta

We take content rights seriously. If you suspect this is your content, claim it here.

Available Formats

Download as PDF, TXT or read online on Scribd

You are on page 1/ 76

Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne

collado Qatreanne collado Qatreanne c

PRELIMS

Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne c
Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne c
BLOOD BANKING & TRANSFUSION MEDICINE transformed into red blood corpuscles”
- Two instances of successful transfusion, both
● Blood Banking: encompasses activities, procedures administered during leg amputation are
and tests done to ensure blood for transfusion is properly documented from the Civil War
collected, preserved, stored and dispensed for later use ● Karl Landsteiner, 1901: discovered the ABO blood
in blood transfusion group system
● Transfusion Medicine: is a multidisciplinary branch of ○ From ABC blood group, naging ABO/ABH blood
medicine that is concerned with the transfusion of blood group
and blood components, including proper selection and ○ O = null or absence of A and B Ag; having the
utilization of blood components as well as removal of most H Ag
blood or blood components in the treatment or ● Von Descatello and Sturli, 1902: discovered the fourth
prevention of disease blood group (Type AB)
● Food and Drug Administration: governing body for ● Karl Landsteiner & Alex Wiener, 1940: discovered Rh
blood bank inspections, inspects blood banks every year. blood group
Blood is regulated both as biologic and as a drug. ● Edward Lindemann, 1913: used vein-to-vein
● Blood transfusion: parang drugs using blood transfusion by using multiple syringes and a special
cannula
HISTORY ● Lester Unger, 1913: designed syringe-valve apparatus;
unassisted transfusion
● CHINA, 1000 BC: the soul was contained in the blood ● Dr. Charles Drew, 1941: director of first American Red
● EGYPTIANS bathed in blood for their health Cross blood bank
● ROMANS drinking the blood of fallen gladiators to gain ● Loutit and Mollison, 1943: introduced the ACD
strength and vitality to cure epilepsy preservative
● Ancient Greeks believed that blood was formed in the ○ WWII = need to preserve
heart, then consumed as it flowed out to the body in ○ Nagkaroon ng research about blood
veins, while air was passed from the lungs to the body in preservatives
arteries ○ ACD preservatives
● Pope Innocent VII, 1492: recipient of first blood ● Robin Coombs, Arthur Mourant & Rob Race, 1945:
transfusion in history describe the use of AHG
● Andreas Libavius, 1615: first person to advocate ● Carl Walter & William Murphy, 1950: introduced the
transfusion, though he is not known to have attempted to use of flexible sealed plastic bag for blood transfusion
perform a transfusion ● Gibson, 1957: introduced citrate phosphate dextrose
● Richard Lower, 1665: is credited with performing the preservative (less acidic)
first successful blood transfusion (animal to animal) ● Herb Cullis, 1972: invented apheresis machine
● Jean Baptiste Denis & Emmerez, 1667: performed ● George Kohler & Cesar Milstein, 1975: discovered
transfusion of lamb blood into the carotid artery of a monoclonal antibodies thru hybridoma technology
young woman ● Dr. Yves Lapierre, 1985: developed gel test in Lyon,
● James Blundell, 1840: performed the first successful France
blood transfusion to patient
○ Clotting was a major problem BLOOD DONATION PROCESS
● Braxton Hicks, 1869: recommend sodium phosphate as
blood preservative ● 500 mL ± 10% of blood (525/110 pound)
● Albert Hustin, 1914: uses sodium citrate as an ● HCT 38%
anticoagulant solution for transfusion ● Anticoagulant Preservative: 63-70 mL
○ Too much citrate = citrate toxicity note:
○ If ↑ citrate = ↓ calcium ● The donor’s RBCs are replaced within 1-2 months after
○ tetani/cardiac arrhythmia donation.
● Richard Lewisohn, 1915: determined the minimum ● A volunteer donor can donate whole blood every 8
amount of citrate needed for anticoagulation and weeks.
demonstrated its nontoxicity in small amounts ● Merong 10-12 pints of blood sa tao; up to 4 pints
● Rous and Turner, 1916: introduced the citrate dextrose ● 16-65 years old pwede magdonate ng blood; if 16,
preservative parent’s consent; if 65, physician’s consent
● Replenish fluid loss within 1-2 mos. (in PH, 3 mos)
Lewisohn’s Method of Transfusion

● 19th Century:
- Saline infusion was observed to be safer than,
and frequently as effective as blood transfusion
- Milk was advocated as potentially effective
infusion because it was thought that the “white
corpuscles of milk were capable of being

CRYOPRECIPITATE ○ 52% protein,

○ 40% lipid,
To treat fibrinogen deficiencies. ○ 8% carbohydrates
Storage Period: 1y in the freezer.

THE DONATION PROCESS

1. Educational Materials
2. Donor Health History Questionnaire
3. Abbreviated Physical Examination

CURRENT DONOR SCREENING TESTS FOR INFECTIOUS

DISEASES:

● Syphilis (1950s)
● HBsAg (1971)
● Anti-HBc (1986)
● Anti-HCV (1990)
● Anti-HIV-1/2 (1992)
● Anti-HTLV-I/II (1997)
● HCV NAT (1999)
● West Nile Virus NAT (2004)
● Trypanosoma cruzi antibody (2007)
● HBV NAT (2009)
● Babesia microti antibody and NAT (2012)
● Zika virus NAT (2016) RBC CHARACTERISTICS:
1. Deformability
HIV = AIDS 2. Permeability
Syphilis = T. pallidum Note:
Western blot - confirmatory test for syphilis sa bata Permeability - ability to pass through
Neurosyphilis - tertiary syphilis; confirmatory test is CSF-VDRL - Permeable substances; water, anions
To monitor HIV therapy - test viral load Freely permeable - readily pass through
Physical examination: RBC is impermeable sa cation
● Blood pressure (systolic, not > 180 mmHg; diastolic, Anion - Cl
not >100 mmHg) Cation - Ca, Na, K
● Temperature, not > 35.5C Na-K pump - ion balance/homeostasis
● Hb, 12.5 g/dL for M, 14-18 g/dL for F Calcium - pump sa interior RBC
● Pulse 50-100 or 60-100 Calmodulin - regulates calcium
- If ↓ ATP = ↑ Ca, Na, kasi hindi gumagana si Calmodulin
RBC BIOLOGY - Nagkakaroon ng rigid cell, not capable of deformability,
and can’t pass through
1. RBC Membrane Deformability - ability of RBC to stretch
2. Hemoglobin Structure and Function - If no ATP, ↓ phosphorylation of spectrin

● Embden Meyerhof Pathway: anaerobic glycolysis that

provides 90% of ATP needed by RBCs
○ Main energy provider
○ Pyruvate kinase
● Pentose Phosphate/Hexose Monophosphate
Pathway: provides 10% of ATP and reduced glutathione
○ Aerobic, prevents Hb oxidation
● Luebering Rapoport Shunt: permits accumulation of
HEMOGLOBIN STRUCTURE AND FUNCTION 2,3-DPG; off-loads oxygen to tissue
● Methemoglobin Reductase Pathway: maintains heme
● Structure: 2 pairs of globin chains, 4 heme groups iron of the hemoglobin in ferrous state
● Normal Types of Adult Hemoglobin:
○ 92-95% HbA = 2 alpha, 2 beta chains
○ 2-3% HbA2 = 2 alpha, 2 delta chains
○ 1-2% HbF = 2 alpha, 2 gamma chains
● Function: gas transport
● Carries oxygen from the lungs to the peripheral tissue;
returns carbon dioxide from the tissue to the lungs for
excretion

HEMOGLOBIN-OXYGEN DISSOCIATION CURVE

RBC PRESERVATION

● Goal: maintain RBC viability during storage

● FDA requires an average 24-hour post transfusion RBC
“Shift to the right” “Shift to the left” survival of more than 75%
○ If natanggal blood, dapat >75% viable
● 70% of red cells must remain viable at the end of the
allowed storage time
● Temperature Requirement:
○ 1-6C: storage temp of liquid whole blood
○ 1-10C: shipping/transport temp of liquid whole
blood
● Storage Lesion (loss of RBC viability):
● Sigmoid shape
○ Increased: hemoglobin, lactic acid, potassium
■ 1 RBC unit = ↑ 1g/dL Hgb and 3% Hct
RIGHT SHIFT LEFT SHIFT ■ Why ↑ Hgb? = RBC breakdown
■ Why ↑ lactic acid? = ↓ oxygen delivery
↑ 2,3-DPG ↑temp., PCO2 ↓ 2,3-DPG ↓temp., PCO2 ■ Why ↑ K? = Na-K pump (leaking)
○ Decreased: pH, glucose, ATP, 2,3-DPG, sodium
↑O2 delivery ↓O2 delivery ■ pH slightly acidic to neutral (RBC)
■ ↓ glucose kasi nacoconsume ng RBC
↓ affinity of ↑ affinity of ■ ↓ 2,3-DPG = if transfused within 24 or
Hb to O2 Hb to O2 8 hours, balik sa normal
● Blood Preservative Contents:
1. Citrate: binds calcium; serves as anticoagulant
↓ pH ↑ pH
a. Removes calcium
2. Adenine: increase ADP levels; for ATP

APPROVED PRESERVATIVE SOLUTIONS:

PLATELET PRESERVATION
HEPARIN: used for by-pass surgery; 2 days shelf-life
CPD – nadedeplete by 2nd week yung 2,3-DPG
CPDA-1 – most common anticoagulant preservative for neonatal PLATELET COMPONENTS:
transfusion - used to treat bleeding associated with thrombocytopenia
CPDA-2 – 42 days; limit (kahit lumagpas pa sa CPDA-2, limit na - transfused prophylactically to increase the circulating
ang 42 days) platelet count in hematology-oncology thrombocytopenic
by-pass surgery – on-going yung surgery tapos binabalik yung patients to prevent bleeding secondary to drug and
blood radiation therapy
● Additive Solutions: support red cell survival up to 42 - Platelets are stored at 20C to 24C with maintaining
days continuous gentle agitation throughout the storage
○ must be added to RBCs within the first 72h of period of 5 days (FDA standards define expiration time
storage as midnight of day 5)
○ added after removal of plasma with/without - Whole blood at room temp kapag hindi pa
platelet gagamitin
○ 100 mL additive solution is used for a 450 mL - Platelets can be stored at 1-6C for 48 hours (but NOT a
blood bag routine practice)
○ Final HCT: 55-65%; (HCT without additive: - Pwede ref temp pero ↓ life span (2 days instead
65%-80%) of 5)
○ Content: saline, adenine, glucose, mannitol or - Expire at midnight of the 5th day (room temp)
saline, adenine, glucose, citrate - pH 6.2; if ↓ pH 6.2 = platelet activation = shape change
● Rejuvenation Solutions: (from discoid to spherical)
○ used to restore levels of ATP and 2,3-DPG - Loss of membrane integrity and function
levels in RBCs stored in CPD, CPDA-1, AS-1
○ Solution can be used up to 3 days after
expiration of red cells (rejuvenation for 1-4 h at
37C)
○ Rejuvesol: only FDA-approved rejuvenation
solution in US; used primarily to salvage
liquid-stored RBCs that have reached out date
○ Content:
Phosphate-Inosine-Glucose-Pyruvate-Adenine; - Platelet concentrate can be prepared from whole-blood
Phosphate-Inosine-Pyruvate-Adenine thru centrifugation or from plateletpheresis.
○ Wash rejuvenated RBCs to remove inosine
○ Freeze rejuvenated RBCs: shelf-life of 10 years RANDOM DONOR PLATELET (RDP)
using CPD and CPDA-1; shelf-life of 3 years
using AS-1 ● Whole blood to be used for preparation of RDP must be
processed within 6-8 hours AFTER COLLECTION (6
● Red Cell Freezing: used for autologous units and hours if ACD; 8 hours for CPD, CP2D, CPDA1).
storage of rare blood types ● Whole blood to be used for RDP preparation must be
○ Storage: 10 years maintained at 20-24C.
■ Frozen RBcs = Thaw at 1-6C = ● Unit should contain at least 5.5x10^10 platelets
remove glycerol = deglycerolization ● Unit must contain sufficient plasma to maintain a pH of
○ Requirement: addition of cryoprotective agent greater than or equal to 6.2 (pH 6.2)
(glycerol) to RBCs that are less than 6 days old ● Platelet units from individual whole blood can be pooled
■ 40% sa autologous units in batches of 6-8 units prior to transfusion.
○ Deglycerolization: done prior to transfusion to ● One RPD unit can increase the blood platelet count by
avoid infusing the hypertonic glycerol 5,000 to 10,000 per uL in a typical 70-g human.
■ Expose to Sodium chloride with
decreasing concentration STEPS:
Red cell washing with decreasing concentration of saline: 12% → 1. Centrifuge whole blood using LIGHT SPIN (3,200 x g for
1.6% → 0.85% (NSS) + 0.2% dextrose 2-3 minutes) to separate the RBCs from the platelet-rich
plasma.

SINGLE DONOR PLATELET (SDP)

● Unit should contain at least 3.0x10^11 platelets per

unit, which is equivalent to 6-10 units of RDP units
● A unit of SDP can increase the blood platelet count by - An increment >10,000/uL is a good increment and
30,000 to 60,000/uL indicates that the patient is not refractory of platelets.
● Platelet count of the donor for plateletpheresis should be - An increment <5,000/uL indicates platelet refractoriness.
higher than 150,000/uL
● Leukocyte-reduced platelets: total leukocytes <5.0x10^6, ● Platelet Refractoriness: failure to yield an increase in
thru specific processing without use of a recipient’s platelet count on transfusion of suitably
leukocyte-reduction filter or by use of a preserved platelets.
leukocyte-reduction filter ● The most common cause of platelet refractoriness is
● 1 SPD unit = 6-10 RDP units HLA alloimmunization
● For patients who have platelet refractoriness, the
PLATELET STORAGE & BACTERIAL CONTAMINATION component of choice is an HLA-compatible SDPs.
● To prevent formation of multiple antibodies to platelets, it
❖ The major concern associated with storage of platelets is best to transfuse platelet units collected via
at 20 to 24 C is the potential for bacterial growth, if the plateletpheresis.
prepared platelets contain bacteria because of ● Platelet Recovery: another method used to check
unremoved contamination at the phlebotomy site or the effectiveness of platelet transfusion
donor has an unrecognized bacterial infection. ○ The expected recovery of platelets is 60% at 1h
❖ RT storage and the presence of oxygen provide a good and 40% at 24h.
environment for bacterial proliferation.
❖ Transfusion-associated sepsis (storage time) NOTES

STORAGE WITH ADDITIVE SOLUTIONS ● Preservatives - added to whole blood to extend shelf life
● Additives - added to RBC with no plasma
ADVANTAGES: ○ If 500 mL blood bag = 110 mL additive
1. Reduces plasma-associated transfusion side effects ○ RBC with no plasma = ↑ Hct
2. Improves platelet storage conditions ○ Reduce HCT from 65%-80%
3. Saves plasma for other purposes (eg, transfusion or ○ Added to RBC to reduce viscosity kasi ↑ Hct =
fractionation) dehydrated
4. Increases the efficiency of procedures for the ○ Saline reduces viscosity para mabilis ang
decontamination of viruses and bacteria transfusion rate
○ Preserves blood for up to 42 days
STORAGE LESION ○ Registered additive in the US:
■ Adsol (AS-1) 42 days
■ Nutricel (AS-3) 42 days
Increased: lactate, degranulation, platelet activation
■ Optisol (AS-5) 42 days
Decreased: pH, ATP, morphology (from discoid to spherical) –
■ Solex
loss of swirling effect, platelet aggregation
● Rejuvenation solution - restore ATP, 2,3-DPG
○ Salvage (save) rare units
FROZEN PLATELETS ○ Extend shelf life of RBC up to 3 days
○ Rejuvesol - FDA approved
● There has been limited use of procedures that freeze ○ 50 mL rejuvenation solution = 1 RBC unit
platelets for clinical use (5% to 6% dimethyl sulfoxide & ○ If exposed ang blood sa open system, dapat
-80C mechanical freezers) ma-transfuse within 24 hours
● One clinical use has been the freezing of autologous ○ Oxygen = ↑ bacterial growth = ↓ shelf life
platelets for subsequent utilization ○ Freeze RBC = 10 years, depende sa
● 2 years shelf life preservative/additive
● Transfusion associated sepsis
○ Platelet concentration is the most common
PLATELET TRANSFUSION blood component associated
○ Most common isolate sa RBC units = Y.
CALCULATING CORRECTED COUNT INCREMENT & enterocolitica
PERCENT RECOVERY

GENETIC TERMINOLOGY ● Serologic testing determines the presence or absence of

antigens on the red cells
● Gene: segment of DNA that encodes a particular protein ● Phenotype: physical expression of inherited traits, is
- A unit of inheritance that encodes a particular determined by reacting red cells with known antisera and
protein and is the basic unit for inheritance of a observing for the presence or absence of
trait hemagglutination
● Chromosomes: structures within the nucleus that contain - The patient’s red cell phenotype is determined
DNA by hemagglutination of red cell antigens using
- Genetic information is carried on chromosomes specific antisera
- 23 pairs: 22 pairs of autosomes and 1 pair of ● Genotype: actual genes inherited from each parent, may
sex chromosomes be inferred from the phenotype
CELL DIVISION allows the genetic material in cells to be - Family studies or molecular tests are required
replicated so that chromosomes can be transmitted to the to determine the actual genotype
daughter cells - DNA-based assays of family studies are
● Mitosis: cell division in somatic cells that results in the needed to determine the genotype
same number of chromosomes
- “all daughter cells are genetically identical to PUNNETT SQUARE
the parent cell”
- The chromosomes are duplicated, and one of ● Examination of family history is an important component
each pair is passed on to the daughter cells in the investigation of inheritance patterns
- During the process of mitosis, quantitatively and ● Punnett Square: illustrates the probabilities of
qualitatively identical DNA is delivered to phenotypes from known or inferred genotypes
daughter cells formed by cell division - Visually portrays the genotypes of the potential
- PMAT: offspring or the probable genotypes of the
1. *INTERPHASE: “resting phase”; cell is not actively parents

● Genes: the basic unit of inheritance on a chromosome SILENT GENES

- Located in specific places called genetic loci
● Alleles: several different forms of a gene; may exist for ● In some blood group systems, genes do not produce a
each locus (for example, A, B, and O are alleles on the detectable antigen product
ABO gene locus) ● These silent genes, known as amorphs, produce
● Antithetical: “opposite”, refers to the antigens produced phenotypes often called null types
by allelic genes (for example, the antigen is antithetical ● The amorphic gene must be inherited from both parents
to the antigen) (homozygous) to produce a null phenotype
● Polymorphic: refers to having two or more alleles at a ● Unusual phenotypes may also result from the action of
given locus, as with the ABO blood group system suppressor genes (act to inhibit expression of another
- The frequency of a particular phenotype in a gene to produce a null expression)
population depends upon the degree of
polymorphism within a blood group system MENDELIAN PRINCIPLES
- A highly polymorphic system makes it less likely
to find two identical individuals (example is one ● Gregor Mendel: Father of Genetics
involving the genes that encode HLA antigens) ● Law of Independent Segregation: refers to transmission
- Because BM and organ transplants require HLA of a trait in a predictable fashion from one generation to
matching, HLA polymorphism contributes to the the next
challenge of finding suitable donors - Illustrates that each parent has a pair of genes
for a particular trait, either of which can be
INHERITANCE PATTERNS transmitted to the next generation
- The genes “segregate” and allow only one gene
● In most cases, blood group antigens are inherited in an from each parent to be passed to each child
autosomal codominant pattern, or with equal expression ● Law of Independent Assortment: demonstrated by the
of both inherited alleles found on autosomes fact that blood group antigens, inherited on different
- Products of each allele is expressed when chromosomes, are expressed separately and discreetly
inherited as a codominant trait ● Law of Dominance: for the two alleles, the dominant
● Recessive: requires that the same allele from both allele is always expressed because it masks the
parents be inherited to show the trait recessive allele
- In blood group genetics, a recessive trait can
express a null phenotype (such as O CHROMOSOMAL ASSIGNMENT
phenotypes) because of homozygosity for a
silent or amorphic gene ● The genetic loci of most of the blood group system
● Dominant: require only one form of the allele to express genes have been determined
the trait (such as group A phenotype that inherits an A
gene from one parent and an O gene from the other
Blood group system chromosome
parent
● Autosomal Dominant: all the members of the family that
Rh 1 Kidd 18
carry the allele show the physical characteristic
- Each individual with the trait has at least one Duffy 1 Lewis 19
parent with the trait
- Present in every generation MNS 4 Landsteiner-Wiener 19
● Autosomal Recessive: not carried on the sex
chromosomes and is carried by either parent or both Chido/Rodgers 6 Lutheran 19
parents but is not generally seen at the phenotypic level
unless both parents carry the trait Kell 7 H 19
● X-linked Dominant: If the father carries the trait on his X
chromosome, he has no sons with the trait, but all his ABO 9 PIPK 22
daughters will have the trait (a father always passes his
Y chromosome to his sons and his X chromosome to his ● Most of the blood group-associated genes are on
daughters) autosomes
- Women can be either homozygous or ● Inheritance patterns are the same, regardless of gender
heterozygous for an X-linked trait, and therefore
when mothers have an X-linked trait, their HETEROZYGOSITY VS. HOMOZYGOSITY
daughters inherit the trait in a manner identical
to autosomal inheritance
● Homozygous: two alleles for a given trait are identical
- The sons have a 50% chance of inheriting the
● Heterozygous: two alleles for a given trait are different
trait or not
● Agglutination reactions vary in strength
- Because the trait is dominant, the sons who
● This variation can be due to the strength of the antibody
inherit it will express the trait
or the density of the antigens on the red cells
● X-linked Recessive: The father always expresses the
● When antigen density varies between red cells of
trait but never passes it on to his sons
different individuals, it is often due to the inheritance of
- The father always passes the trait to all his
the antigen expression
daughters, who are then carriers of the trait
● An individual who inherits different blood group system

Screen for antigen-negative donor units when antisera

LINKAGE AND HAPLOTYPES are unavailable

● In some blood group systems, antigens are encoded by Donor antigen screening for prevention of
two or more genes close together on the same alloimmunization
chromosome and are inherited from each parent as a
unit HDFN Determine parental RhD zygosity
● Genes that are so close together on a chromosome that
Type fetal blood
they are inherited as a unit are linked
- Independent assortment does not occur when
Donor Testing Detect virus in donors that may be below detectable
genes are linked (haplotypes) levels by antibody detection methods
- Haplotypes occur in the population at a different
frequency than would be expected if the genes Relationship Testing Establish paternity and legal relationships for
were not linked immigration
● Linkage Disequilibrium: refers to the phenomenon of
antigens occurring at a different frequency in the ● NAT: a general term used for molecular-based methods
population, depending on whether they were inherited by of screening for infectious agents
linked or unlinked genes - This methodology further reduces the “window
- Also occurs in the HLA system because these period” in viral testing when antibodies may be
genetic loci are very close below detectable levels
- The MHC genes that encode HLA antigens are
inherited as haplotypes
PCR
POPULATION GENETICS
●
The genes encoding many of the red cell and HLA
antigens are sequenced and cloned, providing “maps”
● Statistical application of genetic principles to determine for the specific nucleotide differences that define each
genotype and phenotype occurrence, which is allele
dependent on the gene frequency ● PCR: an in vitro technique used to amplify specific DNA
sequences of interest thru cycles of denaturation,
RELATIONSHIP TESTING annealing of primers that select the area of the DNA for
amplification and replication that result in a millionfold
● The high degree of polymorphism of the HLA and blood copies of a specific area of DNA
group systems has made them valuable tools in cases of Components of PCR:
disputed paternity a. Target DNA: DNA that contains the region of the DNA
● Direct Exclusion: exclusion of paternity when a child has fragment to be amplified
a trait that neither parent shows→ the child has inherited b. Taq polymerase: thermostable enzyme that catalyzes the
a genetic marker that is not found in the mother or replication of template DNA into copies
alleged father c. Primers: short pieces of a single-stranded DNA that are
● Obligatory Gene: gene that is passed by the father to complementary to the opposite strands that flank the
establish probability of paternity (to prove paternity) target DNA
● Indirect Exclusion: failure to find an expected marker in a i. Primers mark the sequence to be amplified and
child when the alleged father is apparently homozygous provide the initiation site on each DNA
for the gene→ the child lacks a genetic marker that the d. Nucleotides: deoxynucleotide triphosphates (dNTPs),

IMMUNOLOGY

BLOOD GROUP ANTIGENS

● are molecules located primarily on the red cell

membrane
● may elicit the production of its corresponding antibody in
individuals who lack the antigen

IMMUNOGENICITY OTHER CHARACTERISTICS

● ability of an antigen to elicit an immune response ● Polyclonal & Monoclonal Antibodies
(immunogens) ○ Polyclonal: produced in response to a single
● Characteristics of Antigens that Determine antigen with more than one epitope
Immunogenicity: ○ Monoclonal: produced by isolating individual B
○ Degree of foreignness cells from a polyclonal population and
○ Molecular size and configuration propagating them in cell culture with hybridoma
○ Antigenic complexity technique; single epitope specificity
● Naturally Occuring and Immune Antibodies: produced in
IMMUNOGLOBULINS & ANTIGEN BINDING reaction to encountered antigens
● Immunoglobulins: are protein molecules that are ● Unexpected Antibodies: all other antibodies directed
produced in response to antigenic stimulation and that against RBC antigens; must be detected and identified
demonstrate specific antibody activity before blood can be safely transfused even if antibodies
react at RT or only with Coombs sera

COMPLEMENT SYSTEM AND BLOOD BANKING

Primary Roles:
A. Direct lysis of cells, bacteria and enveloped viruses
B. Assist with opsonization to facilitate phagocytosis
C. Produce peptide fragment split products, which are
capable of mediating inflammatory and immune
responses such as increased vascular permeability,
smooth muscle contraction, chemotaxis, migration, and
adherence
BLOOD GROUP ALLOANTIBODIES & AUTOANTIBODIES
1. Classical Pathway
● Alloantibody: reacts with a foreign antigen not present on ● Initiate classical pathways- antigen antibody complex,
the patient’s own erythrocytes IgM and IgG
● Autoantibody: reacts with an antigen on the patient’s ○ It starts with C1qrs and up to C4 and C2 to form
own cells C4b which is C3 convertase use it as natural
substrate to make C5 convertase, then C5 is
IMMUNOGLOBULINS SIGNIFICANT FOR BLOOD BANKING used to make a MAC, which will end to cell lysis
● IgG, IgM & IgA: have the most significance for the blood 2. Alternative Pathway
bank ● Initiate alternate pathway- bacterial cell wall, viral, fungi
● Most clinically significant antibodies react at body ○ Start with C3 (unstable so the water hydrolyzes
temperature (37C), are IgG isotype, and are capable of the bond in C3. Once activated, C3b will bind to
destroying transfused antigen positive RBCs and factor B acted on factor B meron ka ng cleaved
causing anemia and transfusion reactions of various product na B
severities 3. Lectin Pathway
● IgM antibodies are most commonly encountered as ● Initiate lectin-maltose, fibrin mismo
naturally occurring antibodies in the ABO system and are ○ Importance of complement system kapag
produced in response to commonly occurring antigens nagkaroon ng distraction duon sa activation it
can result to the damage which is the RBC
IMMUNOGLOBULIN VARIATION damage either intravascular or extravascular
● Isotype: or class variation; refers to variants present in all
members of a species, including the different heavy and
light chains and the different subclasses
● Allotype: present primarily in the constant region; not all
variants occur in all members of a species
● Idiotype: determines the antigen-binding specificity (or
complementary determining regions) of antibodies and
T-cell receptors; found only in the variable (and

INTRAVASCULAR HEMOLYSIS
● usually caused by antibodies directed against the ABO
antigens
● rarely, other IgM blood group antibodies, as well as
some complement-fixing IgG antibodies can induce
intravascular hemolysis RH BLOOD GROUP SYSTEM
● ISBT 004
EXTRAVASCULAR HEMOLYSIS ● the most complex red cell antigen system in humans,
● IgG antibodies cause the majority of extravascular encompassing some 50 antigens, many phenotypic
hemolysis via the RE system, which removes variants and complex serologic relationships
complement-coated RBCs ● D Antigen: most immunogenic of all the Rh antigens
● Rh antibodies do not fix complement (RBCs are almost
ERYTHROCYTE ANTIGENS & ANTIBODIES always cleared through extravascular destruction)
○ Rh because first study in rhesus monkeys

ANTIGEN-ANTIBODY REACTIONS

CHARACTERISTICS

INTERMOLECULAR BINDING FORCES

● Intermolecular binding forces such as hydrogen bonding,
electrostatic forces, van der Waals forces and
hydrophobic bonds, are all involved in antigen-antibody
binding reactions

ANTIBODY PROPERTIES
● Affinity: strength of a single antigen-antibody bond
produced by the summation of attractive and repulsive
forces
● Avidity: a measure of functional affinity of an antiserum
for the whole antigen (combination of affinities); used to
express the binding strength of a multivalent antigen with
antisera produced in an immunized individual
○ Measure the overall stability of antigen antibody
complex
○ If wala kang DAF antigen, you’re prone to
malaria

HOST FACTORS
ABO BLOOD GROUP SYSTEM ● are important in the host’s overall immune response as
● ISBT 001 well as various specific immune reactions
● single most important blood group for the selection and
transfusion of blood
● two antigens (A and B) and four phenotypes (A, B, AB Properties of the host that influence immune response
and O)
● ABO antigens are detectable at 5-6 weeks in utero ● Nutritional status ● Race
● ABO antibodies are weak or absent in the sera of ● Hormones ● Exercise level
newborns until 3-6 months of age ● Genetics ● Disease
○ Diba in blood typing meron tayong forward and ● Age ● Injury
reverse

TOLERANCE
● lack of an immune response or an active
immunosuppressive response

DETECTION OF RBC ANTIGEN-ANTIBODY REACTIONS

BLOOD SAMPLES REQUIRED FOR TESTING

● Serum
● Plasma
○ Principle of BLOOD BANKING is heme
agglutination

FACTORS THAT INFLUENCE AGGLUTINATION REACTIONS

• Centrifugation • Temperature
- induces agglutination - IgM room temp or
reactions, one of the lower, IgG body MONOCLONAL VS POLYCLONAL REAGENTS
important in blood temp 37C
bank • Ig Type
● Monoclonal: are directed against specific epitopes and
• Antigen-Antibody Ratio - ABO (IgM), RH (IgG)
therefore are a potential solution
- zone of equivalence, • Enhancement Media
● Polyclonal: directed against multiple epitopes found on
if there is antigen - Ficin- fig plant
the original antigen used to stimulate antibody
excess - pro-zone, - Papain- papaya
production and can have multiple activities
antibody excess - - Trypsin- pig stomach
pre-zone, inhibition - Bromelain
that causes false -pineapple NEW AND NONTRADITIONAL LABORATORY METHODS
decrease • Chemical Reduction of IgG
• Effect of pH & IgM Molecules FLOW CYTOMETRY
- it should be similar to ● makes use of antibodies that are tagged with a
the serum sample na fluorescent dye
6.5-7.5 ● Fluorescence: occurs when the compound absorbs light
energy of one wavelength and emits light of a different
wavelength
○ Cells that are coated with fluorescent-labeled
antibody emit a brightly fluorescent color of a
specific wavelength
● the principle of flow cytometry is based on the scattering
of light as cells are bathed in a fluid stream thru which a
laser beam enters

SOLID-PHASE ADHERENCE TEST

● can be used to identify both antigens and antibodies
● use antigen-antibody reactions and adherence to a solid
phase support system as opposed to hemagglutination
in solution to determine if a reaction is positive or
negative

GEL TEST
● uses a type of gel matrix instead of saline to separate
positive and negative reactions
● may be used for cell antigen detection and identification,
serum antibody detection and identification, and
crossmatching and is applicable to automated blood
bank methods

RBC AFFINITY COLUMN TEST

● similar to the gel test in that it uses a plastic microtube
filled with a gel-like substance

DISEASES IMPORTANT IN BB SEROLOGIC TESTING

● Immunodeficiency
● Hypersensitivity or allergy
● Monoclonal and polyclonal gammopathies
● HDN
○ erythroblastosis fetalis

1952, Dacie observed that different reaction patterns were obtained when
dilutions of AHG were used to test cells sensitized with warm
(IgG, 37C) as compared with cold antibodies (IgM, <22C)

1957, Dacie & showed that reactivity of AHG to cells sensitized with warm
coworkers antibodies resulted from anti-gamma globulin activity, whereas
anti non-gamma globulin activity by cold antibody

2 major types:
IgM - it binds to corresponding antigen and directly agglutinates
RBC suspended in saline because of its large pentameric
structure
- Pinaka efficient sa agglutination reactions kasi malaki
yung structure niya
IgG - non agglutinating or incomplete antibody due to its single
monomer structure
- It’s too small to directly agglutinate the sensitized RBCs
- AHG test - to detect IgG antibodies and in addition to
IgG, the complement proteins which attached to the red
cells did not produce visible agglutination (?)

AHG TEST

● also called the Coombs’ test

○ Named after Robin Coombs
● first developed by Coombs but principle was
described by Carlo Moreschi
● involves red cells coated with incomplete
antibodies (IgG), which are agglutinated by
NOTES antiIgG in antiglobulin serum thru linking of the
IgG molecules on neighboring red cells
Blood group system ● AHG/Coombs Reagent: green
● Antigen present for
○ A-A
○ B-B
○ O - none
● Mitosis - daughter cells are identical sa parent cells.
● Diploid = 2N
● Haploid = 1N

P – Pull (Chromatin pulled to equatorial plate)

M – Middle (when it sets)
A – Away (mahihila sa isa’t-isa) 1st pic: RBC with antibodies - sensitized RBCs; yung IgG
T – Two (gagawa ng 2 daughter cells) component ng AHG reagent serves as a link sa antibodies para
magkaroon ng visible agglutination
● Antithetical - opposite antigens encoded at 2nd pic: if DAT, the patient sample that is sensitized in vivo +
● Polymorphic: Blood group system ABO, Rh (pinaka reagent (no need for incubation step kasi sensitized na yung RBC
polymorphic in vivo (in the body))
● Genes that encodes the antigen - MHC If IAT, the patient sample can be an antibody, then add red
● ABO genes are located in Chromosome 9 cell antigen reagent OR patient sample is the red cell, then add
● Rh located at Chromosome 1 antibody reagent (there is a need for incubation time because it
needs sensitization before adding the AHG reagent. Upon the
addition of reagent, magkakaroon ng formation of latticework with
visible agglutinates)
ANTIGLOBULIN TEST Principle:
● Antihuman globulins obtained from immunized
HISTORY nonhuman species bind to human globulins such as IgG
or complement, either free in serum or attached to
antigens on RBCs.
● The test uses a reagent which has been prepared by
1945, Coombs, use of antiglobulin test for detection of weak and non injecting animals, usually rabbits. Sheeps and goats may
Mourant, & agglutinating Rh antibodies in serum be used if large volumes of antibodies ang kailangan
Race
○ Yung rabbit, tutusukan ng human antibody

AHG test can be used to detect RBCs sensitized with:

1. IgG alloantibodies - formed during transfusion pag
na-expose sa foreign antigens
2. IgG autoantibodies - formed against self antigens sa
autoimmune diseases
- The presence of autoantibodies causes
problems in the test procedures, usually (+)
DAT results
- DAT is performed to see if may transfusion
reactions na nag-occur during the transfusion or
if may HDN
- (+) DAT = transfusion rxn, HDN, or
hemolytic anemia = hindi compatible
yung nasalin na blood sa patient
- Mahirap hanapan ng compatible na
blood pag (+) DAT
3. Complement components - during blood transfusion,
foreign blood cells enter the body and in the presence of
alloantibodies to antigenic polymorphism on the recipient
tissue, the complement system can be activated, which
can result in adverse transfusion reaction
Polyclonal - made from several different B cell clones that
Two AHG Techniques:
secrete antibodies of different specificity and recognizes different
1. Direct Antiglobulin Test (DAT): test is used detect
epitopes
antibodies or complement bound to RBCs in vivo
Monoclonal - made from a single B cell clone that secrete
2. Indirect Antiglobulin Test (IAT): test is used to detect the
antibodies of the same specificity and recognizes a single epitope
reaction of antibody and RBCs in vitro after an
- Derived sa hybridoma technique
appropriate incubation phase
PREPARATION OF AHG
AHG REAGENTS & PROCEDURES
● AHG production involves injecting human serum or
1. Polyspecific AHG Reagents: produced by hyper
purified globulin into laboratory animals such as rabbits.
immunizing animals, usually rabbits, with purified Ig or
● The human globulin behaves as foreign antigen, the
complement to produce hightitered, high-avidity IgG
rabbit’s immune response is triggered, and an antibody
antibodies
to human globulin is produced.
a. Contains both anti-IgG and anti-C3d antibodies
and detects both IgG and C3d molecules on red
cells
b. Usually ginagamit sa DAT
c. Specific sa IgG and C3b or C3d
2. Monospecific AHG Reagents
a. Reagents are prepared by separating the
specificities of the polyspecific AHG reagents
into individual sources of anti-IgG and
anti-C3d/anti-C3b

METHODS
Application of IAT in the immunohematology laboratory
DIRECT ANTIGLOBULIN TEST (DAT)
● One-stage procedure (patient sample + reagent)
Procedure Purpose
● Ordered to detect IgG or complement proteins bound to
patient cells, which is a consequence of certain clinical Antibody Detects antibodies with specificity to red cell antigens
events, including AIHA, HDFN, drug-related mechanism, screening
or an antibody reaction to transfused red cells
● A positive DAT (agglutination) is an important indicator of Antibody Identifies specificity to red cell antibodies
potential immune-mediated red cell destruction in the identification
body; (-) DAT no agglutination
Crossmatch Determines compatibility between donor and patient before
● Specimen: collected in EDTA transfusion
● DIFFERENTIAL DAT: test that uses monospecific
anti-IgG and monospecific anti-C3d/anti-C3b reagents to Antigen typing Identifies a specific red cell antigen in a patient or donor
determine the cause of a positive DAT with polyspecific
antiglobulin reagents
Comparison of DAT and IAT procedures

DAT IAT

Detects IgG- and complement-coated Detects IgG- and complement-coated

red cells red cells

IgG attachment to red cells has IgG attachment to red cells has
occurred within the patient’s body occurred during the incubation step

One-stage procedure Two-stage procedure

Clinical examples causing a positive direct antiglobulin test Patient’s red cells are tested with Test requires an incubation step before
antiglobulin reagent without an the addition of antiglobulin reagent
incubation step
Clinical condition Caused by Source of IgG

Test for certain clinical conditions: Used as a reaction phase of several

Transfusion Donor cells coated with IgG Recipient (patient)
HDN, hemolytic transfusion reaction, tests in immunohematology: antibody
reaction antibody
and autoimmune hemolytic anemia screen and antibody identification
panel
HDN Fetal red cells coated with IgG Maternal antibody
crossing the placenta

Autoimmune IgG or C3 on patient red cells Patient autoantibody

hemolytic anemia

Drug-related IgG-drug complex attached to Immune complex

mechanism cells formed with drug

INDIRECT ANTIGLOBULIN TEST (IAT)

● Two-stage procedure kasi it includes the incubation step
○ Incubation allows time for the antibody to attach
to the RBC para maging sensitized
● Designed to detect in vitro sensitization of red cells

cannot be dispersed

Common sources of false-negative error in antiglobulin testing

False-negative Possible explanation

Failure to wash cells adequately Unbound human serum globulins

during the test procedure before the neutralize AHG reagent
addition of AHG reagent

Testing is interrupted or delayed; AHG Bound IgG or complement molecules

reagent is not added immediately after may detach from the coated red cells
washing

Failure to identify weak positive Technical error in testing

reactions

Loss of reagent activity Improper reagent storage, bacterial

contamination, or contamination with
human serum

Failure to add AHG reagent Technical error in testing

FACTORS AFFECTING ANTIGLOBULIN TEST
Improper centrifugation: under Conditions for promotion agglutination
1. Ratio of Serum to Cells 6. Saline for Washing centrifugation are not optimal
- Increasing the ratio - Saline should be fresh
increases sensitivity of with suggested open Inappropriate red cell concentrations – Concentration of red cells influences
the test system expiration of 30 days, red cell suspensions fall outside the the agglutination reaction
2. Reaction Medium and buffered to a pH of optimal 2-5%
- Enhancement media 7.2-7.4
[Albumin, LISS, - If stored for long
polyethylene glycol periods in plastic
(PEG)] containers = decreased MODIFIED & AUTOMATED ANTIGLOBULIN TEST
3. Temperature pH = increase the rate TECHNIQUES
- Optimal temp. 37C of antibody allusion
(also for complement during washing process
activation) = false (-)
SOLID-PHASE TECHNOLOGY
- IgG reacts best at body 7. Addition of AHG ● May be used for performing antiglobulin tests
temp - Add AHG immediately ● One of the test reactants (either antigen or antibody) is
4. Incubation Time after washing to bound to a solid phase support (usually a microtiter well)
- Using saline - 30-120 minimize chances of before the test is started
mins removing the
- Using LISS - 10-15 antibodies from the red
mins cell
5. Washing of RBCs 8. Centrifugation for Reading
- At Least 3 times yung - Recommended for
washing before adding evaluation for AHG,
AHG reagent 1000 RCF for 15-20
- Wash to remove secs
unbound proteins or
antibodies

Albumin - rarely used

Mechanism of LISS - reduction of Zeta potential or natural
repulsive effect ng red cell kaya mas mabilis yung agglutination
- decreased incubation time from 30-60 mins to 10-15 mins
PEG - remove water molecules surrounding the RBC thereby
effectively concentrating antibodies
GEL TEST
SOURCES OF ERROR IN ANTIGLOBULIN TESTING ● Detects RBC antigen-antibody reactions by means of a
chamber filled with polyacrylamide gel
● The gel acts as a trap; free non agglutinated RBCs form
pellets in the bottom of the tube, whereas agglutinated
Common sources of false-positive error in antiglobulin testing RBCs are trapped in the tube for hours
● NEGATIVE reactions appear as pellets in the bottom of
False-positive Possible explanation
the microtube; POSITIVE reactions are fixed in the gel
Red cells are agglutinated before Potent cold reactive body of patient origin
washing step and additions of anti
human globulin reagent

Use of dirty glassware Particles or contaminants

Improper centrifugation – over Red cell button packed so tightly on

centrifugation centrifugation that nonspecific clumping

AHG METHODOLOGIES

1. Saline Tube testing

2. LISS Tube testing
3. PEG Tube testing
4. Gel
5. Solid Phase

QUALITY CONTROL OF AHG TEST

● To standardize antiglobulin sera and to confirm true

negative antiglobulin reactions, two types of QC RBCs
are normally used:
○ those coated with IgG
○ those coated with C3b and/or C3d
● To sensitize RBCs with IgG, Rh antibodies are usually
used
● RBCs coated with C3b are prepared by incubation of
whole blood in LISS or with human anti-Le(a) or anti-I
● C3d-coates RBCs are prepared by incubating
C3b-coated with fresh serum or trypsin to split
C3b→C3d
● IgG or complement-coated control cells should give a 1+
to 2+ reaction when tested with anti-IgG or
anti-C3b+C3d

BLOOD GROUP ANTIGEN

● They are a major consideration in solid organ and bone
marrow transplantation because they are widely
expressed.
● They are found on many tissues, body fluids, RBCS,
platelets and endothelial cells.
● We have two antigens in the ABO blood group system,
the A&B antigens.
● 4 phenotypes groups:

A, B AB
Autosomal O - autosomal
codominant recessive
antigen

● Ang – ABO
● Mens- MNS
● Po ni – P1PK ABO FORWARD TYPING (FRONT OR DIRECT TYPING)
● Rhea - Rh
● Lumabas - Lutheran ●Specimen: patient RBC
● Kaya - Kell ●Reagent: uses commercial antisera, anti-A (blue color),
● Lang - Lewis anti- B (yellow), clear anti sera which is for anti D (RH
● Di - Duffy type)
● Kita ni - Kid ● Use: detection of ABO antigens
● Diego - Diego ○ these are required for to be performed on all
donors and patients
ABO BLOOD GROUP SYSTEM (ISBT 001) ○ most frequently performed test in the blood
bank
● The ABO blood group system was discovered in the ○ ABO forward typing or front or direct typing, ang
1900s by Karl Landsteiner which marked the beginning dinedetect natin here are ABO antigens.
of modern blood pumping and transfusion medicine. Therefore, if antigens ang dinedetect, specimen
● Most important blood group for the selection and for patient is RBC or red blood cells.
transfusion of blood. ○ forward typing we perform for either slide (no
● Most common cause of HTR and HDN - most common grading method) or tube method (applicable
cause ABO incompatibility but most severe cause is RH grading).
incompatibility We usually perform the Tube method for initial testing, pwede

Take note, blood banks are the

most critical sections in the lab
because it's a matter of life and
death. Small fault can cause the
death of a patient. Let's say cross
matching, if not careful enough in
the procedure, incompatible blood
might be transfused that could
complicate your patient and
worse could lead to death.

REVERSE TYPING (BACK, INDIRECT OR SERUM TYPING)

● Specimen – serum or plasma

● Reagent – A cells, B cells
● Use – detection of ABO antibodies
○ We detect ABO antibodies, therefore specimen
natin ay serum or plasma the agent
○ reverse typing, no slide method

● graded as 1 +, 2 +, 3 + and 4+ yung reactions. Depende

kung gaano ka weak or strong yung agglutination which
varies from patient to patient.
● first 4+ grading so both A and B cells = O. This is
reverse grouping, meron antibodies to A1 and B cells
● next 0 reactions A1 cells then 3+ in B cells = A
● We possess antibodies antigens na wala tayo since type
A siya meron siyang A antigen wala siyang B antigen so
meron siyang anti B antibody

O gene
- it's considered an amorph kasi walang detectable na
antigen ang naproproduce in response to its inheritance
serving.
- group O is recessive in expression. Then we need 2
alleles to express
- most common blood group types- group O&A
- rarest blood type = group AB

● A1 and A2
○ These are additional phenotypes for Group A

NOTE:
Inversely - relationship between forward and reverse typing
20 seconds - centrifugation time – hemagglutination

● Hemolysis, which is considered as the strongest positive

reaction that can occur and indicates presence of potent
complement fixing antibody and similar doon sa ibang
grading yung 1+, 2+, 3+ and 4 +. 4 then we have the
action.
● Rouleaux, it's not a specific aggregation appearing like a
stack of coins and disappears naman with the addition of
saline.

MIXED-FIELD AGGLUTINATION
● may look like small to large agglutinates with non
agglutinated cells
● may also appear as a “halo” or “puff of smoke” of non
agglutinated RBCs as the RBC button is dislodged from
the test tube bottom
● Causes – receiving non-ABO- type specific RBCs, ABO
subgroups (A3) and bone marrow or HSC
transplantation

● ABO genes - Chromosome 9

● A and B genes - dominant
● Gene
○ Amorph/silent; recessive; no A and/or B antigen
is produced
● inheritance information on ABO antigens
○ It requires for you each antigen, which is
inherited independently of the ABO blood group
system antigens.
● H gene
○ It controls production of H antigen that is
located on a different chromosome from the
ABO genetic locus and in addition on ABO and
H gene, we also have soluble ABO antigens
which is influenced by inheritance of Se gene
● SE gene
○ it will determine new acting secretory status.
○ influences the formation of ABO antigens in
saliva, tears and other body fluids. Sa mga
secretions
○ chromosome 9 located on ABO
○ chromosome 19 = H gene.
● A, B, and H antigens are formed from the same basic
precursor material (called the paragloboside) to which
sugars are attached in response to specific enzyme
transferases elicited by an inherited gene, depende sa
kung anong gene ang present sa kaniya

MUST STUDY

FORMATION OF A AND B ANTIGEN

● Immunodominant sugar
○ Sugars that occupy the terminal positions of this
precursor chain and confer blood group
specificity
○ Therefore, L fucose dito siya ang responsible
for each specificity that is blood group O
○ L fucose it must be formed for the other sugars
to be attached in response to an inherited A
and/or B gene
● H antigen= precursor structure on which the A&B
antigens are made

NOTES
● Genes do not actually code for the production of
antigens but rather produce specific glycosyltransferase
● The H antigen is actually the precursor structure on
which A and B antigens are made
● The H and Se genes are not part of the ABO system;
however, their inheritance does influence A and B
antigen expression
● Without fucose, no other immunodominant sugar will be
attached
● Amount of H (from greatest to least):
O>A2>B>A2B>A1>A1B
● Frequencies: O>A>B>AB

A SUBGROUPS

● So the precursor substance on erythrocytes is referred to

as type 2
● This means that the terminal galactose on the precursor
substance is attached to the North acetyl glucosamine in
the beta 1 to 4 linkage move tile

● Group A phenotype is described by Von Dungen

● In our Anti sera A1 yung naka indicate because it’s the specific
component
● it will not bind-to lectin because it is a plant extract, they bind to
carbohydrates portion of red cell antigen that cause
agglutination
NOTE: lectin is not antibodies and sugar binding origin.
● In routine ABO typing both A1 and A2 red cells react or
agglutinate with anti A reagent. This red cell distinguished with
which is Dolichos biflorus extracted in the seed of plant Dolichos
biflorus
● Dolichos biflorus is specific to A1 and agglutinate but not in A2
red cells
● In routine ABO donor and recipient testing no need to use the
anti-A1 lectin to know what subgroup of A is present for
transfusion purpose.
● We used lectin in resolving typing problems.

● The D galactose is attached in N acetyl glucosamine and

a beta one to four linkage
NOTE!
● type 1 precursor substance refers to beta 1 to 3 linkage
naman between the galactose and N acetyl
● 1-8% of A2 produces anti-A1; 22-35% of A2B produces anti-A1
glucosamine. ● The very potent gene A1 creates between 810,000 and
1,170,000 antigen sites on the adult A1 RBC, whereas inheriting
an A2 gene results in production of only 240,000 to 290,000
antigen sites on the adult A2 RBC
● Dolichos biflorus: source of anti-A1 lectin
● Four forms of H antigen: H1, H2, H3, H4
● H1 and H2: unbranched straight chain
● H3 and H4: complex branched chain

Special techniques of adsorption and elution may be necessary to

demonstrate the presence of A antigen. B SUBGROUPS
● Adsorption: procedure that uses red cells (known antigens) to
remove red cell antibodies from a solution (plasma or antisera);
group A red cells can remove anti-A from solutions
o Antibodies should bind with red cells to remove in the
plasma
o Perform in antibody identification
● Elution: procedure that dissociates antigen-antibody complexes
on red cells; freed IgG antibody is tested for specificity
o Yung mga antibodies na naattach sa red cells naalis yun
o We perform this in drug elution reaction or HDN Phenotypes Description
Mixed field agglutination with anti-B and /or
Phenotypes Description anti -Ab
B3
Mixed field agglutination with anti-A and ● B phenotype is most frequent in weak B
/or anti -Ab phenotype
A3 ● Mixed field define as small Bx Agglutination with anti AB (wk/0 with anti B)
agglutinate within predominantly an
agglutinated cell No agglutination with Anti-B and Anti- AB,
Bm secretor demonstrate quantities of B
Ax Weak agglutination with anti-AB only substance in saliva
<10% red cell shows very weak No agglutination with Anti-A and Anti-AB,
And mixed-field agglutination with anti-A and Bel secretors contain only H substances and no
anti- AB substance in saliva
No agglutination with Anti-A and Anti-AB,
Am secretors demonstrate quantities of A ● Bandeiraea simplicifolia: source of anti-B lectin; used to
substance in saliva agglutinate B cells that have been enzyme pretreated
No agglutination with Anti-A and Anti-AB, o Now we used Griffonia simplicifolia
Ay secretors contain small amount of A ● Anti-AB: used to identify weak subgroups of A and B
substance in saliva ⮚ has a higher titer compared to anti-A or anti-B alone
No agglutination with Anti-A and Anti-AB, ⮚ colorless reagent
Ael secretors contain only H substances and
no substance in saliva LECTINS
● They are all important also called weak subgroup usually Anti-A1 Dolichos biflorus
unnoticed yung reaction nila na need ng further testing para ma Anti-B Griffonia(previously Bandeiraea)
identify. simplicifolia
● A3, Ax, and And show agglutinate but weak Anti-H Ulex europaeus
● Am,Ay, Al no agglutination
ABO HISTOBLOOD AND SOLUBLE ANTIGENS
OTHER SUB A GROUPS ● ABO antigens are present in all tissue and organs of the body
● May be expressed in secretions depending on secretor status

Secretor Gene (Se)

● SeSe or Sese (80%) – secretor
● sese (20%) – non secretor
- Don't expressed soluble H substance in secretion

Detection of Secretor Status

● Specimen: saliva
Ax donor magkakaroon ng reaction with anti A and anti B let’s say ● Principle: hemagglutination inhibition
2+ 2+
Body Fluids Associated with ABH Substances
A B A1 B
● Saliva
O O O 4+ 4+ ● Tears
A O O O O ● Urine
● Digestive juices
● Bile
Simillar yung kanilang reverse typing which is Type O if ni transfuse ● Milk
sa patient na Type O magkakaroon ng reaction Paano? Group O ● Amniotic fluid
meron siyang anti A and B antibodies which agglutinates and lyses ● Pathological fluids: peritoneal, pleural,
Ax RBCs, it will cause intravascular hemolysis. ● pericardial, ovarian cyst

Anti- Anti- A B Anti-H

A b cells cells Lectin
Blood
- - + + +
type O
Oh - - + + -
Source of anti -H lectin :Ulex europaeus

It occurred for when unexpected reactions occur in forward and

reverse grouping.
These are common sources of technical errors resulting in ABO
discrepancies.
Take note that our clerical errors is our most common source of error
ABO discrepancies are classified into four major categories

This is an example of Group 1 discrepancy. Sample from

elderly patient. In forward typing, it shows Group B, but in reverse
grouping no reaction. Supposed to be, reaction is in A1 cells.
Group II: Weakly Reacting or Missing Antigens
● Least common discrepancy
● These are associated for with unexpected reactions in the
forward grouping due to weekly reacting or missing antigens.
● In Group I is associated with reverse grouping due to weekly Here's an example of our Group 3 discrepancy caused by rouleaux
reacting or missing antibodies WHILE Group 2 is associated formation
with forward grouping due to weekly reacting or missing Patient has Group A but the results of forward and reverse grouping
antigens. has my weak reaction on anti B and A1 cells.
Causes include: When determining ABO discrepancies, You look for weak reactions
like 1+, 2 +, it means there’s extra antigens in patient or subgroups
● Subgroups of A & B. of antigens that may react. Sa mga weak reactions lang naman
● Leukemia- It may yield weakened A or B antigens questionable ang results and indicates extra antigens.

● Hodgkin's lymphoma - It's been reported now it can mimic REMEDY

depression of antigens such as seen in leukemia ● Wash rbcs with saline several times to remove these unbound
● Excess blood group Specific soluble substances (BGSS) or extra proteins.

● Acquired B phenomenon - group A individuals na may na ● In the case of Wharton's Jelly- wash 6 to 8 times.
acquire na B Group IV: Miscellaneous Problems
● Antibody to low incidence antigens ● These discrepancies between forward and reverse groupings
are due to miscellaneous problems

Example caused by cold autoantibodies

Remedy
RBC sample
● Wash with saline at 37°C then retype
● Enzyme treated with 0.01 M dithiothreitol to disperse igm related
agglutination
Serum sample
● Warm at 37°C then read results at 37°C = prewarm technique

RH BLOOD GROUP SYSTEM

Note: there are 61 Rh antigens as of the latest update
● Most complex red cell antigen system in humans
● Second most important blood group system in terms of
transfusion (first most important is the ABO blood group
system)
● The most important blood group system associated with
HDN
○ Because it causes severe complications
compared to other blood group systems, ABO
specifically
● The second leading cause of HDN (first is ABO)
● The type of hemolysis is delayed and extravascular
● Rh Function: maintain structural integrity of RBC and
carbon dioxide transporters Fisher-Race: CDE
● D genes are located on the first locus
GENETICS AND BIOCHEMISTRY ● C genes are located on the second locus
● E genes are located on the third locus
● RH genes are inherited as codominant alleles Wiener
○ Codominant - both have equal expressions; ● All alleles are located on one locus
they are both dominant. Ex. C,c (these are Rh Current Rh genetic theory
antigens) ● Located on 2 loci
■ In Rh, when we say “codominant”,
even if one of the antigen is recessive

Fisher-Race: CDE Terminology ● Describes the presence or absence of an Rh antigen

● The Rosenfield system was developed to communicate
● It is used to name the antibody found in a patient phenotypic information more suited for computerized
specimen data entry; it does not address genetic information
● Rh blood group system antigens were inherited as a ○ Unlike Wiener and Fisher-Race that are based
gene complex or haplotype that codes for three closely on genetic concepts, Rosenfield does not
linked sets of alleles address genetic information, it just describes
○ D gene: inherited at one locus the presence or absence of Rh antigen
○ C or c: inherited at the second locus ● D = Rh1
○ E or e: inherited at the third locus ● C = Rh2
● Order of immunogenicity: D > c > E > C > e ● E = Rh3
● c = Rh4
Wiener: Rh-Hr Terminology ● e = Rh5
○ Fisher-Race to Rosenfield equivalent
● It is used to identify red cell phenotypes on the antibody ○ ex. DCe (Fisher-Race) = D+, C+, E-, c-, e+ (in
identification panel cells. etc. Rosenfield, it will be written as 1, 2, 3- ,4- ,5)
○ Wiener believed that there was one gene ○ ex. dce = 4,5 = 1-, 2-, 3-, 4, 5
responsible for defining Rh that produced an
agglutinogen containing a series of blood International Society of Blood Transfusion
factors
● Alleles at one gene locus were responsible for ● Standardized Numeric Terminology
expression of the Rh blood group system antigens on ● Assigned a six-digit number to each blood group
red cells specificity
● Each parent contributes one Rh gene ○ First three numbers: represent the Rh system
● The inherited form of the gene may be identical ○ Remaining three: correspond the Rosenfield
(homozygous) to or different (heterozygous) from each system
parent

● R – denotes the presence of D antigen; (+) D

● r – denotes the absence of D antigen; (-) D
● 1 – presence of C antigen
● 2 – presence of E antigen
● Z and Y – presence of C and E antigens
● ‘ (single prime) – presence of C antigen
● R0 = Dce – most commonly found in black population
● “ (double prime) – presence of E antigen
● R1 = DCe – most commonly found in “WAN” (Whites,
● rh’ – presence of C antigen
Asians, americaNs) populations
● rh” – presence of E antigen
● hr’ – presence of c antigen
● hr” – presence of e antigen SAMPLE PROBLEM 1

The following are reactions using Rh antisera:

Anti-D = positive, Anti-C = negative, Anti-c = positive, Anti-E =
positive, Anti-e = negative
= DcE

What is the probable phenotype of the individual?

A. R1 = DCe
B. R2 = DcE
C. Rz = DCE
D. r = ce or dce

Both parents are heterozygous D/Rh (genotype Dd). What is/are

the possible Rh type of the alisprings?

D d

D DD Dd

d Dd dd
D – presence of D antigen; Rh (+)
d – absence of D antigen; Rh (-)
A. Rh + only
B. Rh - only
C. Both Rh + and Rh -
D. Rh null

Note: Weak D – expressed as Dd (heterozygous); not detected

using anti-D reagent, that’s why we use the AHG reagent
● -D- (D-deletion) – most concentrated; most D antigen because weak D has a strong reactivity with it.
sites
● R1r’ or R0r’ – least D antigen sites

D ANTIGEN

● The most immunogenic antigen in the Rh blood group

system
○ Second most immunogenic is the Kell antigen
● Because of the high immunogenicity of the D antigen,
testing for the presence or absence of the D antigen is
included in routine typing along with the ABO antigens

● The D antigen is made of antigenic subparts genetically

determined that could be absent in rare instances
● If an individual lacks one or more pieces of epitopes of
the total D antigen, alloantibody can be made to the
missing fractions if exposed to RBCs that possess the
complete D antigen
● It is partial D because there’s anti-D to the missing part
● During transfusion, reaction can occur, especially if the
donor has complete D antigen. The recipient with partial
D will produce antibodies to the antigens that they lack

● Control should be negative ● Among the three variations of weak D, only the partial D
● In tube 3, both anti-D and control are positive, hence it is can produce/create antibodies
invalid; unable to interpret the results ● In genetic weak D, Rh protein is complete, but few in
● Tube 1 is Rh (+) because there is a reaction with anti-D number that’s why it is weakly expressed
● Tube 2 is Rh (-) because there is no reaction with anti-D ● In position effect, Rh protein is complete
● In partial D, there’s anti-D to the missing part that’s why
WEAK D this is the only variation that can create antibodies
against the antigens of the donor with complete D
Genetic (weak D)
● D antigens expressed appear to be complete, but few in ● Weak D Testing: also performed on prenatal evaluations
number and Rh immune globulin (RhIG) workups
Position effect (Gene interaction effect) ○ It is required for donors who initially phenotype
● In individuals showing the gene interaction weak D, the as D-negative
allele carrying D is trans or in the opposite haplotype to ○ In case of blood transfusion or donation, if the
the allele carrying C patient is weak D, their Rh type is reported as
● Ce gene is inherited in trans to the RHD gene (C trans to Rh (-); if the donor is weak D, their Rh type is
D) Rh (+)
● ex. Dce/dCe (trans): DCe/dce (cis) ■ Rh (-) blood can be transfused to both
Rh (-) and Rh (+) recipient; that’s also
why the universal blood donor is O-
■ During emergency transfusion (the
patient is in critical condition and is in
need of transfusion), O- blood doesn’t
react and is the safest blood that can
be transfused
■ If Rh (-) blood is transfused to an Rh
(-) recipient, there will be no reaction
because both the donor and recipient
don’t have D antigen
■ If Rh (-) blood is transfused to an Rh
(+) recipient, it is also acceptable
Why because the donor doesn’t have D

Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne c
Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne c
antigen that could react to the SAMPLE PROBLEM 7
recipient’s anti D
■ Rh (+) blood can only be transfused to A patient tests positive for weak D but also appears to have anti-D
Rh (+) recipient in his serum. What may be the problem?
■ If Rh (+) blood is transfused to an Rh A. Mixup of samples or testing error
(-) recipient, transfusion reaction will B. Most weak D individuals make anti-D
occur because Rh (+) donor has D C. The problem could be due to a disease state
antigen and Rh (-) recipient doesn’t, D. A D mosaic may make antibodies to missing antigen parts = if
therefore, since the recipient is now an individual lacks part of the antigen, they may create antibodies
exposed to the D antigen, the recipient to the missing parts if exposed to the whole D antigen
will then create antibodies against the
donor which results in transfusion RH ANTIBODIES
reaction
■ If the weak D patient or recipient is ● Most are lgG and reacts optimally at 37 C
mistyped as Rh (+), they will be ● They are immune antibodies – they are formed if
assumed to be compatible with Rh (-) exposed to foreign antigens
and Rh (+) blood, however, this is not ● Do not bind complement
safe for transfusion because let’s say ● Causes extravascular hemolysis
the Rh (+) blood is transfused, and ● Causes delayed, extravascular HTR
that weak D patient is partial D, the ● Rh antibodies are usually produced following exposure
risk is the recipient will create of the individual's immune system to foreign RBCs, thru
antibodies to the missing antigens, either transfusion or pregnancy
which then leads to transfusion ● HDFN caused by Rh antibodies is often severe because
reaction; to ensure safe blood the Rh antigens are well developed on fetal cells, and Rh
transfusion, if the Px is weak D, their antibodies are primarily lgG which readily cross the
Rh is typed as Rh (-) so they can also placenta
receive Rh (-) blood
■ If the donor is weak D, their Rh is
typed as Rh (+) because Rh (+) blood
is only compatible with Rh (+) recipient
■ If the weak D donor is mistyped as Rh
(-), the blood could be transfused to
Rh (+) and Rh (-) recipients, which is
not okay because the donor has D
antigen which can cause transfusion
reaction for the Rh (-) recipient
○ In weak D testing, for safe blood transfusion,
the blood of the donor and recipient should be
compatible; weak D donor – Rh (+), weak D
recipient Rh (-)
○ In weak D testing, the results are not reported
UNUSUAL PHENOTYPES
as weak D (+) or Du (+); it’s either Rh (+) or Rh
(-) only
○ Weak D testing is also performed in evaluating D-Deletion Phenotype
the Rh immune globulin (Rhogam)
● RhIG: is purified anti-D prepared from immunized donors ● -D- or D-
and is given to D-negative mothers to prevent the ● Phenotype whose red blood cells possesses an
formation of anti-D extremely low number of D antigen that most reagent
○ Composed of anti-D antibodies and is given to anti-D are unable to detect
D-negative mothers to prevent the formation of ● Due to mutation of RHD gene
anti-D once the fetal RBCs leaked because of a ● Occurs most often in individuals of Southeast Asian
tear in the placenta or during delivery descent, occuring in o. up to 30% of that population
○ Rhogam is tested with weak D to ensure that ● Adsorbing and eluting ant:-D from the individual's RBCs
the mother is Rh (-) is often the only way to detect the D antigen
○ This procedure requires incubating ant:-D with
SAMPLE PROBLEM 6 the RBCs in question at 37 C followed by
eluting the anti-D off the adsorbed RBCs
Positive agglutination in anti-A antisera in forward typing
Positive agglutination in B cells in reverse typing. Rh null Phenotype
A. A+ = could be a possible answer, however, there was no Rh
typing performed ● -/-
B. A ● No Rh antigens; null – lack of Rh antigens
C. B+ ● Cells that type as Rh null have membrane abnormalities
D. B that shorten their survival and cause hemolytic anemia of
varying severity
● The inheritance of the Rh null phenotype can result from
a regulator gene or an amorph gene
● When transfusion of individuals with Rh null syndrome is
necessary, only Rh null blood can be given

Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne c
Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne c
○ In Rh null, it has universal antibody to all Rh produce antibodies against the antigens
antigen ● There are Rh antigens that are not frequent and are not
● Individuals with Rh null syndrome demonstrate a mild strongly expressed in the red cells; you’ll only know that
compensated hemolytic anemia, reticulocytosis, they are present once a reaction occurs
stomatocytosis, a slight-to-moderate decrease in ● Antigen-negative RBCs should be transfused if
hemoglobin and hematocrit levels, an increase in HbF, a antibodies to Rh blood group system antigens identified
decrease in serum haptoglobin, and possibly an elevated or have been previously noted in the patient's history.
bilirubin level ● It is important to check previous records of patients who
○ Stomatocytosis is a characteristic of Rh null may be transfused for a history of red cell antibodies that
phenotype may have developed from previous transfusions or
pregnancies
Rh mod Phenotype
Hemolytic Disease of the Fetus and Newborn
● Individuals of the Rh mod phenotype have a partial
suppression of Rh gene expression and exhibit features ● Due to Rh incompatibility results from Rh (-) mother and
similar to those with Rh null syndrome, however, the Rh (+) fetus or newborn
clinical symptoms are usually less severe and rarely ● Initially observed in infants of D-negative women with
clinically remarkable D-positive mates
○ Compared with Rh null, Rh mod phenotype is ● First pregnancies were usually unaffected. Infants from
less severe subsequent pregnancies were often stillborn or severely
● Red cells lack most of their Rh antigen expression anemic and jaundiced
because of the inheritance of a modified RHAG gene ○ First pregnancies were usually unaffected
○ Unlike Rh null, Rh mod does not completely because there is no sufficient time for the
lack Rh and LW (Landsteiner-Wiener) antigens mother to produce enough anti-Rh antibodies to
● Hemolytic anemia is also a characteristic of this harm the fetus
phenotype ○ Once the mother has produced enough anti-Rh
antibodies, the fetus’ RBCs will be destroyed
CLINICAL CONSIDERATIONS ● The initial pregnancy stimulated the mother to produce
anti-D from the exposure to D-positive cells that occurred
Transfusion Reactions during birth when the infant's and mother's circulations
mixed
● Rh antigens are highly immunogenic; D antigen is the ● Because maternal anti-D antibodies can cross the
most immunogenic outside ABO system placenta, fetal red cells in subsequent pregnancies were
● Exposure to the antigen when the antibody has formed destroyed by the mother's antibody
produces a rapid secondary immune response ● RhoGAM (Rh immune globulin) – used to prevent HDN
● Rh mediated HTR usually results in extravascular ○ Contains antibodies against Rh antigens and
destruction of immunoglobulin-coated RBCs – delayed can be injected or given to the mother during
extravascular HTR pregnancy, before delivery, or immediately after
○ The transfusion recipient may have an delivery, miscarriage, or abortion
unexplained fever, mild bilirubin elevation, and ○ These injected antibodies will bind to fetal
a decrease in hemoglobin and haptoglobin RBCs that may have entered the mother’s
○ HTR is detected using DAT (+) blood and inactivate them; injected antibodies
○ When the DAT indicates that the recipient’s will inactivate fetal Rh antigens and prevent
RBCs are coated with IgG, elution studies may sensitization of the mother
be helpful in defining the offending antibody ● If HDN due to ABO incompatibility occurred, it is less
specificity severe compared to Rh HDN, and most commonly
○ Elution – dissociates antigen-antibody caused by type O mother and type A baby;
complexes on red cells; freed IgG antibody is ○ all children with ABO HDN are affected – baby
tested for specificity usually appears jaundiced (elevated bilirubin)
○ In the case of HTR, there is an antigen-antibody ○ Treatment for ABO HDN is phototherapy where
complex because there is a reaction; the they are exposed to light to convert bilirubin to
foreign antigen of the donor binded to the photo bilirubin, then it will be excreted in the
recipient’s antibody which is shown through the bile;
DAT. With elution, it will specify which antibody ○ Exchange transfusion
causes the transfusion reaction, and the
antibody that will be recovered in the elution LW BLOOD GROUP SYSTEM (ISBT 016)
procedure is called the ELUATE
○ If an antibody was recovered either in the ● Originally identified as anti-Rh in early experiments
serum from the DAT or eluate from the elution involving rabbits immunized with rhesus monkey blood
technique, the implicated antigen that caused ○ Identified as Rh because it agglutinated first in
the reaction should not be present in the Rh (+), then in Rh (-) which should not be it
subsequent transfusions ● Anti-LW is different from anti-D
○ ex. CC patient transfused with O- blood, and ● Ant-LW agglutinates Rh+ and Rh- cells except Rh null
the patient is A-, there is no problem because (--/--)
the patient is compatible with the blood type, ○ Anti-D – agglutinates/reacts with Rh (+) only
however, transfusion reaction can still occur if ● Anti-LW more frequently appears as an autoantibody,
the patient lacks the antigen that was which does not present clinical problems
transfused in them because then their body will

LEWIS BLOOD GROUP SYSTEMS (ISBT 007)

● Produced by tissue cells (found in secretions)

● Not well developed at birth
● Lewis gene (Le, FUT3) codes for the production of
fucosyltransferase enzyme
● Unique because Lewis antigens are not intrinsic to RBCs
but are type 1 glycosphingolipids that are passively
absorbed on the RBC membrane plasma

Lewis Antigens

● The Le(a-b-) phenotype is found more frequently among

Africans The Le(a+b+) phenotype is rare among whites
and Africans but is more frequent among Asians

P1PK BLOOD GROUP SYSTEM (ISBT 003)

● The P blood group was introduced in 1927 by

Landsteiner and Levine
● In their search for new antigens, they injected rabbits
with human RBCs and produced an antibody, initially
called anti-P, that divided human RBCs into two groups:
P+ and P-
● The P blood group antigens, like the ABH antigens, are
synthesized by sequential action of glycosyltransferases,
which add sugars to precursor substances
● The precursor of P1 can also be glycosylated to type 2H
chains, which carry ABH antigens
● P1, P or Pk may be found on RBCs, lymphocytes,
granulocytes, and monocytes
● P can be found on platelets, epithelial cells, and
fibroblasts
● P and Pk have also been found in plasma as
glycosphingolipids and as glycoproteins in hydatid cyst
fluid
● The RBC antigens of the P blood group exist as
glycosphingolipids The P blood group antigens are
resistant to treatment with ficin & papain, DTT,
chloroquine, and glycine-acid EDTA. Reactivity of the
antibodies can be greatly enhanced by testing with
enzyme-treated RBCs

Lewis Antibodies

Disease Association
● Glycophorin A may serve as the receptor by which
certain pyelonephritogenic strains of E. coli gain entry to
the urinary tract
Disease Association ● The malaria parasite Plasmodium falciparum appears to
● The P system antigens also serve as receptors for use alternative receptors, including GPA and GPB
P-fimbriated uropathogenic E. coli- a cause of urinary
tract infections I BLOOD GROUP SYSTEM (ISBT 027)
● The Pk antigen is a receptor for Shiga toxins, which
cause Shigella dysentery and E. coli-associated HUS
● P is the receptor of human parvovirus B19
● Pk provides some protection against HIV infection of
peripheral blood mononuclear cells

MNS BLOOD GROUP SYSTEM (ISBT 002)

I and i Antigens Reactivity

Disease Association
● Anti-I is associated with CAD and M. pneumoniae, and
anti-i is associated with IM
● Conditions associated with increased i antigen on RBCs
include those with shortened marrow maturation time or
dyserythropoiesis: acute leukemia, hypoplastic anemia,
megaloblastic anemia, sideroblastic anemia,
thalassemia, sickle cell disease, PNH, and chronic
hemolytic anemia
● Chronic dyserythropoietic anemia type II or hereditary
erythroblastic multinuclearity with a positive acidified KIDD BLOOD GROUP SYSTEM (ISBT 009)
serum test (HEMPAS) is associated with much greater i
activity
● In Asians, the i adult phenotype has been associated
with congenital cataracts

DUFFY BLOOD GROUP SYSTEM (ISBT 008)

KELL ANTIBODIES

KELL BLOOD GROUP SYSTEM (ISBT 006)

KELL ANTIGENS
● Excluding ABO, K is rated second only to D in terms of
immunogenicity
● The Kell blood group system consists of 36
high-prevalence and low-prevalence antigens; it was the
first blood group system discovered after the introduction
of antiglobulin testing
● Can be detected on fetal RBCs as early as 10 weeks
and is well-developed at birth
● Kell blood group antigens are found only on RBCs. They
have not been found on platelets or on WBCs.
● The Kell glycoprotein is covalently linked with another
protein, called Xk, by a single disulfide bond. Kell antigen
expression is dependent upon the presence of the Xk
protein.
● The absence of Xk results in McLeod syndrome. It is
also associated with CGD.
● The prevalence of K antigen is low, and the chance of LUTHERAN BLOOD GROUP SYSTEM (ISBT 005)
receiving a K+ unit is small.
● Blood bankers seldom deal with the serology of the
Lutheran blood group system. The antigens are either
high prevalence, so only a few people lack the antigen
and can make an alloantibody, or vert low prevalence, so

Prevalence of Most Common Lutheran Phenotypes

PHENOTYPE MOST POPULATION (%)

Lu(a+b-) 0.15

Lu(a+b+) 7.5

Lu(a-b+) 92.35

Lu(a-b-) Very rare

The Lu(a-b-) Phenotype

COLLECTION PROCEDURE

AABB: American Association of Blood Banks

● was established in 1947
● an internal association of blood banks that includes
hospital and community blood centers, transfusion and
transplantation centers, and individuals involved in
transfusion medicine
● Mission: to establish and provide the highest standard of
care for patients and donors in all aspects of transfusion
medicine

FDA: US Food and Drug Administration

● inspects blood banks on an annual basis
● blood is regarded as a biologic and a drug
● 1988, Center for Biologics Evaluation and Research
(CBER): responsible for regulating the collection of blood
components used for transfusion and for the
manufacture of pharmaceuticals derived from blood and
blood components develops and enforces quality
standards, inspects blood establishments, and monitors
reports of errors, accidents, and adverse clinical events
● 1990s: FDA began to treat blood establishments such as
manufacturers, requiring strict compliance toward all
aspects of transfusion medicine including donor
selection and screening responsible for licensing of
reagents and blood products as well as blood center
inspections.

DONOR SCREENING

REGISTRATION
● Blood collection facilities must confirm donor identity and
link the donor to existing donor records.
● Most facilities require photographic identification such as
a driver’s license or school identification card.
● The following is a list of information used by the
collection facility in the registration process and is kept
on record by use of a single donation record form:
○ Name (first, last, MI)
○ Date and time of donation
○ Address
○ Telephone
○ Gender
○ Age or date of birth
○ Consent to donate

MEDICAL HISTORY QUESTIONNAIRE

● Obtaining an accurate medical history of the donor is
essential to ensure benefit to the recipient.
● The interviewer should be familiar with the questions,
and the interview should be conducted in a secluded
area of the blood center.
● The medical history is conducted on the same day as the
donation.

PHYSICAL EXAMINATION
General Appearance
● observe donor for presence of excessive anxiety, drug or
alcohol influence, or nervousness
Weight
● maximum of 10.5 mL/kg of donor weight for WB
collection

INFORMED CONSENT
● The donor must be informed of the risks of the
procedure, and also of the tests that are performed to
reduce the risk of infectious disease transmission to the
recipient.

AUTOLOGOUS DONORS
ACUTE NORMOVOLEMIC HEMODILUTION:
● Autologous Donor: one who donates blood for his or her
● A blood conservation technique that involves removal of
own use (donor-patient)
whole blood from a patient with infusions of crystalloid or
● Safer than allogeneic: no risk of disease transmission,
colloid before surgical blood loss
transfusion reactions, or alloimmunization to white blood
● The blood is collected in standard blood bags containing
cells, RBCs, platelets, or plasma proteins
anticoagulant/preservative and stored at RT
● Four Different Types of Autologous Donation:
● The “shed” blood is normally reinfused to the patient
1. Preoperative collection
within 8h of collection (viability of platelets and
2. Acute normovolemic hemodilution
coagulation factors)
3. Intraoperative collection
● Blood should be properly labeled and must contain the
4. Postoperative collection
patient’s full name, medical record number, date and
time of collection, and the label For Autologous Use Only
PREOPERATIVE COLLECTION:
● Blood units for acute normovolemic hemodilution (ANH)
● indications include patients undergoing orthopedic
are reinfused in the reverse order of collection so that
procedures, vascular surgery, cardiac or thoracic surgery
the last unit reinfused carries the highest hematocrit level
and radical prostatectomy
● The maximal surgical blood order schedule can provide
guidance for surgical procedures to estimate the number

Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne c
Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne c
INTRAOPERATIVE COLLECTION: Height 5’1” 5’5”
● Involves collecting and reinfusing blood lost by a patient
during surgery Hematocrit Minimum of 40% Minimum of 40%
● Most programs use devices that are capable of collecting
the shed blood, washing it with saline, and then
concentrating the RBCs with hematocrits in the range of 5. Stem cell pheresis*
50-60%
● The vacuum setting on the device should be <100 torr to ALLOGENEIC DONOR DEFERRAL
guard against possible hemolysis of recovered blood ● Temporary: prospective donor is unable to donate blood
● Contraindicated when procoagulants are applied to the for a limited period of time
surgical field ● Indefinite: prospective donor is unable to donate blood
● Used in cardiothoracic, major orthopedic and cardiac for an unspecified period of time
surgery and vascular surgeries such as liver ● Permanent: prospective donor will never be eligible to
transplantation donate blood (these donors may be eligible to donate
autologous blood)
POSTOPERATIVE BLOOD COLLECTION:
● Blood is collected from a drainage tube placed at the 1 MONTH ● German measles (rubella) vaccination
surgical site DEFERRAL ● Chicken pox vaccination
● It is reinfused with or without processing via a ● Drug:
● Isotretinoin (Accutane) – acne treatment
microaggregate filter to screen out any debris ● Finasteride (Proscar) – for benign prostate
● This blood is characterized as being dilute, partially hyperplasia
hemolyzed,and defibrinated
1 YEAR ● Rape victim
DIRECTED DONORS DEFERRAL ● After HBIg administration
● Healthcare worker exposed to blood or body fluids
● Directed Donation: directed toward a specific patient ● Tattoo
● Needle stick injury
PHERESIS DONATION ● Ear or body piercing
● Sexual contact with a prostitute, with AIDS,
1. Plateletpheresis hepatitis, with hemophiliacs or with IV drug users
● Donation frequency: every 2d (no more than 2x in 7d; no ● Female who had sexual contact of men who ever
more than 24x in 12 mos) had sex with another men
● Platelet count: at least 150,000/uL or 150x10^9’L ● Travel to endemic areas with malaria (3 years –
immigrants, refugees, citizens who resided on
● Donor should not take medication that interfere with endemic areas and diagnosed/infected with
platelet function: malaria)
○ Aspirin – 48h deferral ● *Philippines:
○ Stayed for less than 6 months: 6 months
○ Feldene – 48h deferral ○ Stayed for more than 6 months/resident: 1
○ Clopidogrel (plavix), ticlopidine (TIclid), year
Ticagrelor (Brilinta), vorapaxar (Zontivity), and ○ Infected: 3 years
prasugrel (Effient) – 14d deferral ● Syphilis/gonorrhea
● Received transfusion of blood components or other
human tissues (organ, tissue, BM transplant, or
2. Plasmapheresis bone/skin graft) known to be possible sources of
● “Occasional”: donor undergoes pheresis not more often bloodborne pathogens
● Rabies vaccination
than once every 4 weeks, and donor selection mimics ● Major operation including dental surgery
that of whole blood collection ● Have been in juvenile detention, lockup, or prison
● “Serial”: donation is more frequent than once every 4 for more than 72h (incarceration in a correctional
weeks, and additional requirements apply institution for 72h)
● Donation Frequency:
PERMANENT/ ● Hemophiliacs
○ Frequent: every 48h (no more than 2x in 7d) INDEFINITE ● IV drug users
○ Infrequent: every 4 weeks (no more than 13x a DEFERRAL ● (+) AIDS
year) ● (+) HBsAg
● Serum total protein should be at least 6g/dL ● (+) HTLV
● Malignant solid tumors
● If the donor weighs 50-80 kg – not more than 500 mL ● Drugs: Etretinate (Tegison) – treatment for psoriasis
WB should be removed at one time ● History of Babesiosis
● If the donor weighs more than 80 kg – not more than 600 ● History of Chagas disease
● Chronic cardiopulmonary
mL WB should be removed at one time ● Renal disease
● Liver disease
3. Leukopheresis ● Recipient of cornea and dura mater transplant
● Special agents required for collection of granulocytes ● Recipient of human pituitary derived GH
● Bovine insulin
from leukopheresis donor: ● Leukemia, lymphoma and myeloproliferative
○ HES disorder
○ Corticosteroids
○ GFs (eg, G-CSF) OTHER ● Tooth extraction: 3 days
DEFERRAL ● MMR vaccination: 8 weeks (NBVSP); 4 weeks
● Childbirth: 6 weeks after delivery (AABB); 9 months
4. Double RBC pheresis after delivery (Ph)
● Donation Frequency: every 16 weeks ● Typhoid, Oral polio, Mumps, Measles (rubeola),
● Specifications: Yellow fever: 2 weeks
● Aspirin or Piroxicam: 48h
MALE FEMALE ● Plavix (Clopidogrel) or Ticlid (Ticlopidine): 14 days
● Individuals with a history of dengue or chikungunya
virus: 6 months following full recovery from
Weight At least 130 pounds At least 150 pounds

infection)
WHOLE BLOOD COLLECTION
● Individuals with TB: 2 years
● Smallpox vaccination: 14-21 days or until scab has DONOR IDENTIFICATION
fallen off
● A numeric or alphanumeric system is used to link the
donor to the donor record, pilot tubes, blood container
NO DEFERRAL ● Toxoids or killed/synthetic viral, bacterial or
rickettsial vaccines such as diphtheria, hepatitis A, and all components made from the original collection.
hepatitis B, influenza, Lyme disease, pneumococcal ASEPTIC TECHNIQUE
polysaccharide, polio injection (Salk), anthrax, ● Iodine compound (such as PVP-iodine or polymeriodine
pertussis, plague, parathyroid, rabies, Rocky
mountain spotted fever, tetanus, or typhoid injection
complex)
if the donor is symptom-free and afebrile ● Area is scrubbed at least 4 cm in all directions from the
site for a minimum of 30 seconds
OTHER ● GH from human pituitary glands – for children with
MEDICATIONS delayed or impaired growth: permanent COLLECTION PROCEDURE
● Insulin from cows (bovine, or beef, insulin) – to treat
diabetes: indefinite
● Feldene – for mild to moderate arthritis pain: 48h or 1. Confirm donor identity, and make the donor as comfortable as possible.
2d
● Aspirin – blood thinner and prevent blood clots;
inhibits platelet function: 48h or 2d 2. Using a tourniquet or blood pressure cuff, select a large, firm vein in the
● Plavix (Clopidogrel) & Ticlid (Ticlopidine) – inhibits antecubital space that is free of any skin lesions or scarring. Inspect both arms.
platelet function used to reduce the chance for
heart attack and stroke: 14d 3. Prepare the site using an FDA-approved cleansing method. When finished,
● Experimental medication or unlicensed cover the site with sterile gauze.
(experimental) vaccine – associated with a research
protocol and the effect in blood donation is
4. Inspect the blood bags for any defects or discoloration.
unknown: 1 year unless indicated by medical
director
● Ticagrelor (Brilinta) & Prasugrel (Effient) – both 5. Ensure the balance system is adjusted to the volume being drawn; ensure
used to prevent blood clots and inhibits platelet counterbalance is level. Place hemostats on the tubing to prevent air from
function; Ticagrelor is used for prevention of stroke, entering the line.
heart attack and other events in people with acute
coronary syndrome, meaning problems with blood 6. Reapply tourniquet or blood pressure cuff (40 to 60 mm Hg) to increase
supply in the coronary arteries: 14d distention of the vein.

7. Uncover the sterile gauze, and perform the venipuncture immediately. Check
MEDICATIONS THAT CAUSE BIRTH DEFECTS the position of the needle, and tape the tubing to the donor’s arm to hold the
needle in place. Cover with a sterile gauze.
DRUG PURPOSE DEFERRAL
8. Release the hemostat, and ask the donor to open and close the hand every 10
to 12 seconds during the collection procedure.
Proscar (Finasteride) For prostate gland 1 month
enlargement
9. Reduce the pressure on the cuff to approximately 40 mmHg.
Propecia (Finasteride) For baldness 1 month
10. Continue to monitor the patient throughout the entire collection process. The
donor should never be left unattended. Mix blood and anticoagulant periodically
Accutane (Amnesteem, Given for severe acne 1 month
during the procedure (e.g., every 45 seconds).
Sotret
Claravis, Isotretinoin)
11. When the primary bag has tripped the scale, the donor can stop squeezing
and tubing can be clamped. A unit containing a volume of 405 to 550 mL should
Avodart (Dutasteride) For prostate 6 months
weigh between 429 to 583 g, plus the weight of the container and anticoagulant.
enlargement
The conversion 1.06 g/mL is used to convert grams to milliliters. If the volume
collected is in the low volume range (300 to 404 mL in a 450-mL collection or 333
Soriatane (Acitretin) For severe psoriasis 3 years to 449 mL in a 500-mL collection), the unit must be labeled as a “low volume
unit,” and fresh frozen plasma (FFP) cannot be made from this unit, as it would
Tegison (Etretinate) For severe psoriasis Permanent not contain adequate levels of coagulation factors.

12. Before the needle is removed from the donor’s arm, pilot tubes are filled. The
pressure is reduced to 20 mm Hg or less and, depending on the tubing of the
bag, the tubes are filled:
SAMPLE PROBLEM 1 ● In-line needle. A hemostat or metal clip is used to seal tubing distal to
the needle. The connector is opened, the needle is inserted into the
The donor weighs 42 kg. Identify the allowable blood to be pilot tube, the hemostat is removed, and tubes are filled. The donor
needle can now be removed.
collected, amount of anticoagulant needed, and amount of ● Straight-tubing assembly. Place hemostats approximately four
anticoagulant to be removed segments from the needle. Tighten the loose knot made previously in
the tubing; release the hemostats, and strip a segment of tubing
between knot and needle. Secure hemostat and cut tubing in a
stripped area of segment. Fill required tubes by releasing hemostats.
This is an open system, and appropriate biohazard precautions
should be followed. Reapply hemostats and remove needle from
donor’s arm.
SAMPLE PROBLEM 2
13. Once the needle has been removed from the donor’s arm, apply pressure
over gauze, and ask the donor to raise his or her arm, continuing to exert
The donor weighs 37 kg. Identify the allowable blood to be pressure over the site. When the bleeding has stopped, the donor can lower his
collected, amount of anticoagulant needed, and amount of or her arm, and an appropriate bandage can be applied.
anticoagulant to be removed.
14. The needle assembly should be discarded into an appropriate biohazard
receptacle. The tubing should be stripped to allow proper mixing of
anticoagulant/preservative with blood.

15. Heat seal the filled tubing into segments, and apply an appropriate
○ Special transfusion requirements
identification label to one segment and detach from the blood bag for storage. ● Test recipient’s serum or plasma to detect
Place blood at the appropriate temperature. Units in which platelets will be made clinically significant antibodies.
must be maintained at room temperature (20°C to 24°C) until the platelet ○ Method must include 37 C incubation
concentrate has been prepared; all others can be stored at 1°C to 6°C.
with conversion to antihuman globulin
using reagent red cells that are not
pooled
Pretransfusion Testing
4. Donor Testing
● Is an entire quality process composed of many ● Confirm ABO and Rh types on donor red cell
procedures designed to provide the safest blood product units as required
possible for the recipient of a transfusion ○ Confirmation testing for ABO group
● Often considered synonymous with crossmatching required on all units (serum testing not
● Clerical error resulting in incorrect ABO groupings: major required)
cause of transfusion-associated fatalities; greatest threat ○ Confirmation of Rh type required only
to safe transfusion therapy on those units labeled as Rh-negative
● Include: (weak D testing is not required)
○ Recipient identification
■ Misidentification of recipient: most 5. Crossmatch
common cause of error ● Select ABO- and Rh-compatible red cell
○ Sample collection and handling components for transfusion
○ Pretransfusion testing ● Perform serologic or electronic crossmatch of
red cell components
STEPS IN PRETRANSFUSION COMPATIBILITY TESTING ○ Full antiglobulin crossmatch required if
current antibody screen is positive or
1. Physician (or other authorized health professional) order the patient has a known history of
● Written (paper or electronic) order for type & clinically significant antibodies
screen test
○ Type and screen 6. Labeling
○ Crossmatch RBCs ● Label all red cell products or other components
○ Special requests such as irradiation, with the recipient’s identifying information
CMV-negative, etc. ○ Label must contain at least two
○ Oral orders may be accepted if independent patient identifiers, donor
followed by the written request unit number, and compatibility test
results, if performed
2. Phlebotomy/Recipient Identification
● Phlebotomist must positively identify the SPECIMEN REQUIREMENTS
recipient and the recipient’s blood sample
○ Transfusion request and labeled RECIPIENT SAMPLE IDENTIFICATION
patient blood sample must contain at
least two pieces of independent ● A physician order is required before blood is drawn for a
identifying info for that patient type and screen
○ Mechanism to identify the date of ○ Order may be verbal (followed by a written
sample and the phlebotomist (paper or order) or may be given on paper or
electronic) electronically
○ Transfusion service will ensure that ● Recipient blood specimens must be labeled by the
identifying information on the phlebotomist at the bedside directly from the information
requisition and sample label is in on the patient’s wristband
agreement ○ If the patient’s wristband is not attached for any
● Two Independent Identifiers: reason, signed confirmation of the patient’s
● Patient’s first & last name identity by nursing staff should be obtained
● Unique ID number (birth date, driver’s before blood is drawn.
license, photographic ID) ○ Label: patient’s FULL name, hospital
● If px does not have a wristband or if px ID is identification number, spx collection date
unknown = temporary ID with a tie tag or
wristband or ankle band - should not be TYPE OF SAMPLE
removed until proper ID has been attached to
the px (confirmed verification of identity) ● Serum: traditionally been preferred for compatibility
testing
3. Recipient Testing ○ Collected in siliconized plain tubes without
● Perform ABO and Rh typing on recipient’s serum separator gel
blood specimen- red cell and serum or plasma ● Plasma: increasingly used owing to newer testing
testing are required. technologies (e.g., gel test)
● Review previous records for comparison with ● Appearance: reject grossly hemolyzed recipient blood
current typing results. samples as unacceptable for pretransfusion testing
○ Clinically significant antibodies (unless there is no other choice)
○ Significant adverse events to ● *grossly hemolyzed or lipemic samples = false positive
transfusion readings with an automated gel tech instrument

ABO GROUPING

ABO GROUPING OF DONORS

● The collecting facility is required to perform a forward

and reverse ABO type on all donors.
● The transfusion service or other laboratory responsible
for compatibility testing must confirm the ABO group of
all units of RBCs or whole blood received.

ABO GROUPING OF RECIPIENTS

● Both cell and serum groupings are required for initial ● Principle: The crossmatch was added to compatibility
ABO testing of all patient samples, and the results of testing to:
RBC and serum testing should agree with each other ○ Prevent life-threatening or uncomfortable
before any results are reported and blood is transfused. transfusion reactions
○ Maximize in vivo survival of transfused red cells
RH GROUPING
CROSSMATCH TESTING: TYPES
RH TYPING OF DONORS
1. Major Crossmatch
● All donor units testing as Rhnegative by direct ● Donor: RBC
agglutination are tested for a weak D phenotype by the ● Patient: Serum
IAT. 2. Minor Crossmatch: to test opposite compatibility
○ If routine D testing or weak D testing is positive, ● Donor: Serum
the unit is labeled Rh Positive ● Patient: RBC
● The transfusion service must reconfirm the D type of all
RBC units labeled Rhnegative by testing red cells from Minor crossmatches are rarely performed, for two main reasons:
an integral attached segment by a direct anti-D 1. Transfused blood is screened for unexpected (non-ABO)
agglutination method. ab, so performing a minor crossmatch to make sure a
○ Repeat testing of donor units by the transfusion non-ABO ab won’t cause a problem doesn’t make a lot
service for weak D is NOT required by the of sense
AABB Standards. 2. Since the volume of transfused plasma is generally small
● IAT: in vitro in comparison to the patient’s blood volume, minor
● DAT: in vivo incompatibilities (even if not detected by the donor ab
● Weak D donor: Rh positive screen) are not of great consequence
● Weak D recipient: Rh negative - For example, a group O red blood cell
transfusion to a group A recipient would have a
RH TYPING OF RECIPIENTS minor crossmatch incompatibility, but is not
contraindicated due to the very small amount of
● Every pretransfusion recipient blood specimen must be incompatible antibody present in the unit of red
tested for the D antigen by direct typing. cells.

Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne c
Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne c
SAMPLE PROBLEM ○ Saline replacement = to resolve rouleaux (true
agglutination will remain)
Predict compatibility for the following situation. Assume that the
patient has a negative antibody screen and autocontrol. INCUBATION PHASE/ LISS PHASE
Patient: B+ ● Done at 37 C
Donor: O+ ● Usage of enhancement media such as LISS
A. Compatible major crossmatch ● Detection of clinically significant antibodies such as IgG
B. Incompatible minor crossmatch
C. Incompatible major crossmatch ANTIGLOBULIN CROSSMATCHING PHASE
● Commercial high-tittered antiserum is required when
CROSSMATCH TESTING: CLASSIFICATION selecting these antigen-negative donor units.
● A reading is required only at the antiglobulin phase
Serologic Crossmatch ● Done at 37 C
● Tests recipient’s serum/plasma with the red cells from ● Several enhancement media may be applied to boost
the donor unit antigen-antibody reactions. They may include albumin,
● Can be performed using two main methods: LISS, PEG and polybrene
hemagglutination (tube and gel testing) and ● Possible reasons for incompatibilities:
solid-phase red cell adherence ○ New alloantibody is present in recipient’s
plasma
Solid-phase red cell adherence ○ Alloantibody to a low-incidence is present on
● Components of antigen-antibody reaction is immobilized the donor unit RBCs
onto a solid medium ○ Warm-reactive autoantibody is present in the
● On centrifugation antigen positive cells spread out while recipient’s plasma
antigen negative cells form a button at the bottom of the ○ Donor unit has a positive DAT
well
● Excess plasma is blotted out and anti-IgG bound CROSSMATCH TESTING: PRINCIPLES & METHODS
indicator red cells are added to give visible reaction
Broad Spectrum Compatibility Test
● Developed to be an all-inclusive test, one which would
detect clinically significant antibodies without sacrificing
speed – so essential in an emergency, or simplicity so
important in prevailing errors.
● Three Phases:
○ Protein Phase
■ Incompatibility in the ABO system will
be detected at this point. Hemolysis
especially may indicate the presence
of an immune anti–A and / or anti–B.
Computer Crossmatch ■ An incompatibility due to cold
● Uses a computer to make the final check of ABO agglutinins.
compatibility in the selection of appropriate units for ■ The prozoning anti–Rh antibodies are
transfusion detected in a serum albumin mixture
on immediate centrifugation.
CROSSMATCH TESTING: PHASES OF CROSS MATCHING ○ Thermo Phase
■ Incompatibility in this phase is usually
due to the presence of a low tittered
IMMEDIATE SPIN/ SALINE OR INITIAL PHASE:
anti –Rh antibody that does not react
● May be used for recipients with no evidence of clinically
on immediate centrifugation.
significant antibody or antibodies in the current sample
■ Certain Rh antibodies (anti–C, anti–E
and in the historical record
and some anti–D) occasionally react
● Recipient serum or plasma and donor red cell
only in an albumin medium and are
suspensions are mixed and observed for agglutination
non–reactive in the antiglobulin test.
● Main purpose: detect ABO incompatibility
○ Antiglobulin Phase
● Done at RT
■ Antibodies are detected here which
● Uses NSS or 22% albumin enhancement media (30
usually react only in this test (anti–Fja,
mins; 30-60 mins*)
anti– Jka, anti–X).
● There should be no delays in the centrifugation step or
■ These antibodies in the Rh system
reading of the reaction. If IS crossmatch is not properly
which react only in the antiglobulin test
performed, false-negative results may occur with failure
(so called “third order” or
to detect ABO incompatibility
“panagglutinoid” antibodies) are noted.
● Possible reasons for incompatible IS crossmatch:
■ Antibodies present in acquired
○ Incorrect ABO grouping of recipient or of donor
hemolytic anemia will be found.
unit selected
○ Cold-reactive allo- or auto-antibody in the
● Abbreviated Crossmatch
plasma not detected in antibody detection tests
○ If a patient has no known history of and / or no
○ Abnormalities in the recipient’s plasma such as
currently demonstrable unexpected antibodies,
rouleaux
ABO and Rh type specific blood is cross
● Remedy: perform saline replacement and repeat IS
matched at the time of need by using an
crossmatch

Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne c
Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne c
immediate spin–saline abbreviated crossmatch. SPECIAL TOPICS
○ Not done in the Ph
○ Combination of type & screen + IS URGENT REQUIREMENT FOR BLOOD & BLOOD
COMPONENTS
Computerized (Electronic) Crossmatch
● Uses a computer to make the final check of ABO If the blood is released before the completion of testing,
compatibility in the selection of appropriate units for the patient’s record must contain the physician’s signed
transfusion statement indicating the clinical situation was urgent and
● Advantages: required release of uncrossmatched blood.
○ Increased time efficiency ● Release signed by physician
○ Reduced volume of sample needed on large ● Tag on donor unit indicating emergency release:
crossmatch orders compatibility or infectious disease testing was not
○ Greater flexibility in staffing completed at the time of issue
○ Better management of blood bank inventory ● Patient name and identifiers
○ Potential for a centralized transfusion service ● Donor unit number(s), ABO and D phenotype, expiration
date
TAGGING, INSPECTING, ISSUING AND TRANSFUSING ● Retain segments from units for crossmatching
BLOOD PRODUCTS ● Name of person issuing units
● O negative blood
● When the appropriate compatibility testing has been ● If the patient DIES as a result of the emergency,
completed and the unit/s are suitable for transfusion, a remaining compatibility testing may be waived
tag is produced and attached to each donor unit. or abbreviated at the discretion of the
● The donor unit tag must clearly state the patient’s full transfusion service physician. Testing should be
name & ID number, name of the product, donor complete enough to show that the death was
number, expiration date, ABO & D phenotype of the unrelated to the transfusion of uncrossmatched
unit, interpretation of cross matching test (if performed), blood.
and identity of the person doing the testing or selection
of unit COMPATIBILITY TESTING FOR TRANSFUSION OF PLASMA
PRODUCTS
REQUIREMENTS FOR THE TAG ON THE CROSSMATCHED
DONOR UNIT ● Compatibility testing procedures are NOT required.
● Intended recipient’s two identification identifiers ● Transfusion of LARGE volumes = crossmatch test
● Unique donor unit number or pool number between the donor plasma and patient RBCs may be
● Interpretation of compatibility tests, if performed performed.
● * If compatibility testing is incomplete or shows ● Primary purpose for testing: detect ABO incompatibility
incompatibility, this information must be indicated in bold between donor and patient (immediate spin crossmatch)
on the tag.
● A physician’s order for blood or a blood product must be INTRAUTERINE TRANSFUSIONS
on file for the transfusion to occur.
● The person requesting the donor unit presents a ● Transfusion of blood into the fetus
transfusion request form to the blood bank staff ● Blood must be compatible with maternal antibodies
indicating the desired product and the intended recipient. capable of crossing the placenta.
● This form is checked carefully and independently by both ● If ABO and Rh groups of the fetus have been determined
persons against the unit tag. The unit tag information is following amniocentesis, chorionic villus sampling or
also checked against the unit label. percutaneous umbilical blood sampling, group-specific
● The expiration date is carefully checked to ensure that blood could be given provided that there is no
outdated units are not issued. fetomaternal ABO or Rh incompatibility.
● The unit is visually checked for discoloration, clots or ● If the ABO and Rh groups of the fetus are not known,
other abnormal appearance. group O Rh-negative RBCs should be selected for the
● * These checks are documented with the name of the intrauterine transfusion.
person issuing the unit and the person picking it up. Date ● Crossmatch testing is performed using the mother’s
& time of issuance and unit’s destination are also serum sample.
documented.
NEONATAL TRANSFUSIONS
REQUIREMENTS FOR THE ISSUE OF BLOOD & BLOOD
COMPONENTS
● Blood for an exchange or regular transfusion of a
● Physician’s order
neonate (younger than 4 months of age) should be
● Intended recipient’s two independent identifiers, ABO
compatible with any maternal antibodies that have
group, and D phenotype
entered the infant’s circulation and are reactive at 37 C
● Donation identification number, donor ABO group, and if
or AHG.
required, the Rh type
○ For both intrauterine and infant (younger than 4
● Interpretation of crossmatch tests, if performed
months) transfusions, blood should be as
● Blood product expiration date, and if applicable, time
FRESH as possible and no older than 7d.
● Special transfusion requirements, if applicable
● Antibody detection testing can be performed using
● Date & time of issue
maternal serum or infant’s serum (e.g., cord serum)
and/or eluate prepared from the infant’s RBCs
○ Removed antibodies from the surface of RBCs

Finalssssssss

“The first and most important step in ensuring that transfused TTI: a disease or disease agent
blood will not transmit a pathogenic virus is careful selection of Two Criteria:
donors.” 1) Disease or disease agent that could be fatal or
- Main goal is safe blood transfusion life-threatening and potentially cause permanent damage
- Logbook – to trace Px if positive for infections or of a body function or body structure requiring medical or
diseases surgical intervention.
- Hematology – NKTI 2) Disease or disease agents may pose a risk of
- Confirmatory screening for STI – San Lazaro Hospital transmission by blood or blood components or by a
- Confirmatory screening for syphilis/TTIs – RITM blood byproduct manufactured from blood or blood
- Hepatitis A, E, G – same cases of transfusion reactions components.
- Hepatitis A – 28 days incubation period/window/core
period (period exposed until showing signs and TRANSFUSION-ASSOCIATED HEPATITIS
symptoms Hepatitis: inflammation of the liver
- Hepatitis C – caused by single stranded RNA; others –
DNA Symptoms
- Hepatitis A, E – fecal-oral
Jaundice Anorexia Nausea
- A – picornaviridae; B – hepadnaviridae; C – flaviviridae;
E – caliciviridae Dark urine Malaise Abdominal pain

REQUIRED TESTING ON ALLOGENEIC AND AUTOLOGOUS Hepatomegaly Fever Vomiting

DONOR BLOOD

● The goal of donor testing is to improve the safety of the Hepatitis A

blood supply. ● Picornaviridae
● Infectious hepatitis or short-incubation hepatitis
Required Donor Blood Tests ● Transmission: fecal-oral (spread thru water, food and
person-to person contact)
Testing for Testing Performed ● Incubation Period: 28 days (peak viremic period occurs 2
weeks before the onset of the elevation of liver enzymes
RBC antigens ABO and D phenotype or the appearance of jaundice
● (+) IgM anti-HAV antibody: required for diagnosis of
Clinically significant Antibody screen
RBC antibodies Hepatitis A
○ IgM antibodies are detectable at or prior to the
Hepatitis HBsAg onset of clinical illness and declines in 3-6
months
Anti-HCV ○ IgG antibodies appear soon after IgM and may
persist for years after the infection
Anti-HBc ● HAV circulates in the bloodstream only during the initial
phase of infection, when an individual is usually too ill to
HCV NAT donate; however, if blood is collected while the virus is
circulating, it can be transmitted by transfusion.
HBV DNA ● Because transfusion transmission is extremely rare,
donated blood is not tested for hepatitis A antigen or
HIV-1/2 Anti-HIV-1/2
antibody.
HIV-1 RNA
Hepatitis B
HTLV-I/II Anti-HTLV-I/II
● Hepadnaviridae
● Serum hepatitis; first known hepatitis virus transmitted by
Syphilis Nontreponemal serologic test for syphilis (eg, blood transfusion
RPR) or IgG or IgG + IgM antibody to T. pallidum ● Transmission: sexual, parenteral, perinatal; exposure to
antigens bodily fluids (parenteral – beyond intestines; perinatal –
before, during, and after birth)
WNV WNV RNA ○ High levels: blood, serum, wound exudates
○ Moderate: semen, vaginal fluids, saliva
T. Cruzi (Chagas IgG antibody to T. cruzi (one-time testing for donor
disease) screening)
○ Low: urine, feces, sweat, tears and breast milk
● Dane particle: complete HBV virion
● HBV DNA: first marker to appear; can be detected in the
Autologous Donation: serum about 21 days before HBsAg and may be useful
● The intended recipient is the blood donor adjunct in detecting early acute HBV infection, and also
● Phenotyped for ABO and D antigens and are tested for to evaluate the effectiveness of antiviral therapy in
unexpected antibodies patients with chronic hepatitis B
● Blood collection facilities are NOT required to perform
infectious disease testing on autologous blood products
that are not shipped.
● A directed donation undergoes the same testing as an
allogeneic donation.

Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne c
Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne c
HBV is sufficient to avoid hepatitis D transmission.
Markers (SEC – Ag; CES – Ab)
Hepatitis E
HBsAg • First serological marker to appear
(surface) • Early indicator of active infection ● Caliciviridae
• Important marker in screening blood donors ● Transmission: fecal-oral (contaminated drinking water in
• Indicates active HBV infection, acute or chronic developing countries)
• Serum HBsAg usually becomes undetectable by 4-6 months
after the onset of symptoms in patients with acute hepatitis B
○ HEV 1 & 2: fecal-oral
○ HEV 3 & 4: foodborne
HBeAg • Present during periods of active replication of the virus ● Incubation Period: 2-6 weeks
(envelope) • Appears shortly after HBsAg and disappears shortly before ● Major cause of hepatitis globally
HBsAg in recovering patients ● Associated with a high rate of mortality in pregnant
• Indicates a high degree of infectivity when present
women
● No FDA-approved serologic methods
HBcAg (core) • Not detectable in serum because of the viral envelope that
masks it
• Detected only through biopsy of the infected liver Hepatitis G
• NOT included in hepatitis profile ● Transmission: bloodborne (parenteral), fecal-oral and
respiratory route, vertical/perinatal
Anti-HBc IgM • First antibody detected
or • Indicator of current or recent infection
● Not a major cause of liver failure
HBcAb IgM • Useful in detecting infection during “core window” period ● RT-PCR: diagnosis of current, ongoing infection
• Not a marker of immunity
• Persists for the lifetime of the individual Hepatitis Tests
• Serves as a lifelong marker of HBV infection
● To prevent the transmission of hepatitis by transfusion,
Anti-Hbe or • Indicates that the patient is recovering from HBV infection
five tests are currently performed on donor blood:
HBeAb • Marker of convalescence and favorite prognosis ○ HBsAg
○ Total anti-HBc (IgM and IgG antibody)
Anti-HBs or • Appears during the recovery period of acute hepatitis B, ○ Antibody to HCV (anti-HCV)
HBsAb weeks to months after HBsAg disappears ○ NAT to detect HCV RNA and HBV DNA
• Persists for years and provide protective immunity
• Also produced after immunization with the hepatitis B
vaccine, which consists of recombinant HBsAg produced from Hepatitis B Surface Antigen
genetically engineered yeasts or mammalian cells ● Screening Tests: enzyme & chemiluminescent
• Protective titers of the antibody in the serum are considered immunoassays
to be 10 mIU/mL or higher
● Confirmatory: uses the principle of specific antibody
neutralization to confirm presence of HBsAg
● The FDA requires donor screening for HBsAg, ○ If HBsAg is present in the serum, the antibody
anti-HBc, and HBV-DNA binds the antigen
● IgM – recent infection ○ The neutralized HBsAg is blocked from binding
● IgG – chronic infection to the antibody coated solid medium
○ If the neutralization causes the reaction to
Hepatitis C disappear or diminish by at least 50%, the
● Flaviviridae original result is considered positive for HBsAg
● “Non A - Non B”
● Transmission: sexual, parenteral, perinatal/vertical Antibody to Hepatitis B Core
● Incubation Period: 2-26 weeks ● “surrogate marker”
○ Average incubation period: 7-8 weeks, followed ● No specific confirmatory test for anti-HBc
by seroconversion occurring in 8-9 weeks
■ Seroconversion – first exposed to Antibody to Hepatitis C Virus
pathogen until the body produces Ab ● Screening Tests: Third-generation ELISAs and
against pathogen chemiluminescent assays (detectable by approximately
● Most frequent cause of chronic liver infection and the 10 weeks after infection)
leading indicator for liver transplantation ● Confirmatory Test: NAT
● Standard treatment involved a combination of pegylated
IFN-alpha and ribavirin Nucleic Acid Testing to Detect DNA of HBV and RNA HCV
● The current recommended screening for DNA and RNA
Hepatitis D uses a mini pool of 6 to 16 donor samples.
● Delta hepatitis ● Sensitive NAT techniques for HCV RNA even in pooled
● Transmission: sexual donor samples have reduced the window period for HCV
● Only found in patients with HBV infection (requires detection to 7.4 days.
HBsAg in order to synthesize an envelope protein and ● HBV DNA testing has reduced the window period for
replicate) HBV detection to 18.5 to 26.5 days.
● Highest risk: IV drug users
● Detected by testing for IgM or IgG anti-HDV or HDAg HIV TYPES 1 AND 2
and HDV RNA in the serum
● Tests for HDV are not required for blood donations
● Although hepatitis D virus is also transmitted by blood,
HIV-1: HIV-2:
the concurrent presence of HBV is necessary to cause
disease. ● First virus designated as the ● Also causes AIDS
● HDV is a defective virus found only in HBV carriers. causative agent of AIDS ● More common in Africa
○ HDV doesn’t exist without HBV ● The virus infects CD4-positive ● Appears to produce a less
● Blood is NOT screened for HDV because testing for

T lymphocytes (helper T cells) severe disease

WEST NILE VIRUS
● Viremia is first detectable in ● Both forms of HIV are spread ● Mosquito Borne flavivirus that manifests symptoms
plasma 10 days to 3 weeks by sexual contact, perinatal ranging from a mild febrile illness to encephalitis, coma,
after infection breastfeeding, and parenteral and death
exposure to blood
● Most donor collection facilities ○ Bird: primary amplifying hosts in a mosquito
use a combination test that bird-mosquito cycle
detects antiHIV-1 and ● Other Route of Infection: blood components, donated
anti-HIV-2
organs, and breast milk
● Screening Test: IgM antibody-capture ELISA (serum &
● Screening Test: EIA (used for the qualitative detection of CSF); NAT
HIV-1/2 in a ChLIA test) ○ Plaque Reduction Neutralization Test: most
● Positive screening tests are repeated in duplicate, and if specific test for arthropod-borne flaviviruses
at least one of the duplicates also tests positive, a and helps to distinguish false-positive IgM
confirmatory test is performed antibody-capture ELISA from cross reactivity
● Confirmatory Tests: HIV-1 indirect immunofluorescence ● Currently, it is NOT logistically feasible for all units to be
assay, HIV-2 ELISA and rapid diagnostic test used for tested individually.
HIV-1 and HIV-2 differentiation
● HIV: CD 4:CD 8 – 2:1 CHAGAS DISEASE
● NV: 1:2 ● Causative Agent: T. cruzi
● American trypanosomiasis
SYPHILIS ● Transmission: contact with feces of infected reduviid
bugs; blood transfusion
● Causative Agent: T. pallidum ○ Infected individuals can experience severe
● Transmission: direct sexual contact; through transfusion heart or intestinal problems
● Was the first infectious disease screening performed on ● Screening Tests: ELISA, chemiluminescent assays
blood donations
● The routine storage of RBCs at refrigerated Malaria, Chagas disease and WNV are referred to as infections
temperatures limits survival of T. pallidum, but platelets transmitted by insect vectors.
stored at RT could transmit the organisms
● Donated blood can be tested by various methods, CYTOMEGALOVIRUS
including RPR, hemagglutination tests, and the FTA-Abs ● Risk of CMV Infection:
test. ○ Highest: individuals receiving allogeneic
● A positive confirmatory test result defers the donor for at marrow transplants and the fetus
least 12 months. ○ Moderate: recipients of solid organ transplants,
persons with HIV, and individuals who may
MALARIA require an allogeneic marrow transplant in the
future
○ Low: low birth-weight neonates and autologous
● Caused by Plasmodium species (P. malariae, P.
marrow recipients
falciparum, P. vivax, P. ovale)
● Leukocyte-reduced or CMV-negative units reduce the
● Transmission: through bite of a female Anopheles
risk of CMV infection in low birth-weight neonates.
mosquito
● Transmission: person to person through contact with
○ But infection may also occur following
infected body fluids (urine, semen, saliva, blood, cervical
transfusion of infected blood
secretions, and breast milk)
● Plasmodium can survive in blood components stored at
● Screening Tests: ELISA, fluorescence assays, indirect
RT or 4C for at least a week, and deglycerolized RBCs
hemagglutination, latex agglutination
can transmit disease.
○ If patient is symptomatic, active infection can be
detected by viral culture of urine, throat swabs,
Other Transfusion-Transmitted Infections/Diseases and tissue samples
● CMV – most frequently transmitted virus from mother to
HUMAN T-CELL LYMPHOTROPIC VIRUSES TYPES I/II fetus
● Blood and blood components are NOT universally
HTLV-I: screened for CMV
● First retrovirus to be associated with a human disease
● Associated with adult T-cell lymphoma/leukemia (ATL) EPSTEIN-BARR VIRUS
● Associated with HTLV-I-associated myelopathy or ● Virus is shed in the saliva and is most frequently
tropical spastic paraparesis (HAM/TSP) associated with infectious mononucleosis.
● Also associated with uveitis & infective dermatitis of ● EBV was first discovered in Burkitt’s lymphoma cells.
children, Sjogren’s syndrome, polymyositis, and facial ● Associated with cancers such as nasopharyngeal
nerve palsy carcinoma, non-Hodgkin’s lymphoma, oral hairy
● HTLV-I/II are transmitted through cellular blood products, leukoplasia in AIDS patients, T-cell lymphomas, and
breast milk, sexual contact, contaminated needles, and Hodgkin’s disease
injection drug users. ● Kissing dse
● The risk of HTLV-I/II transmission through transfusion is
less than 1:2,000,000 units transfused. ZIKA VIRUS
● Screening Test: HTLV antibody testing ● Transmission: mosquito borne (Aedes aegypti and
● No FDA-approved confirmatory test for HTLV (Western Aedes albopictus) – intrauterine, perinatal,
Blot, however, is commonly used as a supplemental test laboratory-acquired, sexual- and transfusion-associated
for confirmation of a positive HTLV antibody result). ● Associated with severe neurological complications

TICK-BORNE BACTERIAL AGENTS

● Lyme disease NOTES
● Rocky mountain spotted fever
● Ehrlichiosis ● Preservatives - added to whole blood to extend shelf life
● Additives - added to RBC with no plasma
TRANSFUSION-ASSOCIATED PARASITES ○ If 500 mL blood bag = 110 mL additive
○ RBC with no plasma = ↑ Hct
Babesia microti ○ Reduce HCT from 65%-80%
● Transmission: bite of an infected deer tick, blood ○ Added to RBC to reduce viscosity kasi ↑ Hct =
transfusion, solid organ transplantation dehydrated
○ Units of packed RBCs (liquid stored and frozen ○ Saline reduces viscosity para mabilis ang
deglycerolized) and platelet units, which contain transfusion rate
RBCs, have been associated with transmission ○ Preserves blood for up to 42 days
○ Registered additive in the US:
Trypanosoma cruzi ■ Adsol (AS-1) 42 days
● Chagas disease ■ Nutricel (AS-3) 42 days
● The AABB TTD Committee has given T. cruzi an orange ■ Optisol (AS-5) 42 days
category rating. ■ Solex
○ An orange category agent is considered a low ● Rejuvenation solution - restore ATP, 2,3-DPG
scientific or epidemiological risk regarding blood ○ Salvage (save) rare units
safety. ○ Extend shelf life of RBC up to 3 days
○ Rejuvesol - FDA approved
○ 50 mL rejuvenation solution = 1 RBC unit
○ If exposed ang blood sa open system, dapat

Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne c
Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne c
The donor RBCS, destroyed by the recipient antibodies, shortening
ma-transfuse within 24 hours then new RBC survival by causing new acting HDR. Clinically
○ Oxygen = ↑ bacterial growth = ↓ shelf life significant antibodies are typically IgG class which react at 37°C that
○ Freeze RBC = 10 years, depende sa is body temperature or say AHG phase of indirect anti globulin test
preservative/additive DAT (Direct Antiglobulin Test)
● Transfusion associated sepsis - work up of HTR, HDN, hemolytic anemias in in vivo
○ Platelet concentration is the most common
sensitization in RBCS
blood component associated
○ Most common isolate sa RBC units = Y.
IAT (Indirect antiglobulin test)
enterocolitica
- blood typing, cross matching, antibody screening, antibody
○ 2nd most common isolate sa RBC units =
identification that is in vitro sensitization of RBCS
Pseudomonas spp.
IgG = 37°C or body temp
○ Most common contaminant sa RBC units =
IgM = room temperature or 22°C or lower (M for Malamig)
Propionibacterium acne
Antibody screen
○ Most common isolate sa platelet = B. cereus,
- The process for detection if there’s unexpected antibody.
S. epidermidis
If positive antibody screen, proceed to antibody identification using
○ Blood in general = S. aureus, S. epidermidis
our antibody panel or antigram.
■ S. aureus - most common cause
■ ↑ deaths due to blood product
ANTIBODY SCREEN
contamination
● purpose is to detect any potential clinically significant antibody
● Blood substitutes in case of blood component shortage:
in a donors or recipients sample.
○ Hgb-based oxygen carriers (HBOCs)
● The AABB standard requires the use of an antibody screen to
○ Perfluorocarbons (PFCs)
detect clinically significant antibodies as part of pretransfusion
testing on the recipient.
Detection and Identification of Antibodies ● Must include incubation at 37°C and the use of antiglobulin test.
Main goal is to identify the irregular or unexpected antibodies as o 3 phases in detection of antigen antibody complexes
opposed to ABO which are expected antibodies because they are - immediate spin
naturally occurring. - LISS or 37°C
- AHG = final reading
Antibody Detection ● Included (but not on AABB requirement) in standard prenatal
● It's crucial in work up or investigation in hemolytic translation testing for obstetric patients in order to evaluate risk of HDN in
reactions, immune hemolytic anemias and also some monitoring fetus and to evaluate the mother's candidacy for Rh immune
patients or mothers are at risk of delivering infants with HDFN globulin (RhIg)
● Also required in donors of allergenic blood and blood products
Antibodies can either be autoantibody or Alloantibody. and stem or progenitor cells.
Autoantibody ● Antibody screens are performed to detect antibodies in the
- antibodies produce own antigens from the word, auto following people:
meaning self o Patients requiring transfusion
Alloantibody o Women who are pregnant after delivery or after delivery
- antibodies produce following exposures of foreign antigens. o Patients with suspected transfusion reactions
Refereed as self-antigens, came from other individual and o Blood and plasma donors
usually we develop Alloantibodies if we are transfused with ● When antibodies are detected in a blood donor, the resulting
blood components will be labeled with the identity of the antibody (with
the exception of cryoprecipitate and wash or frozen or
ANTIBODIES deglycerolize RBCS)
● Immune Alloantibodies
- These are produced in response to RBC stimulation
through transfusion, transplantation or pregnancy.
Transplantation of cornea - It does not require cross matching
unlike your liver and kidney.
This is an example of antibody screen red cell anti-gram. Nagkaroon
● Naturally occurring alloantibodies ng positive antibodies screen may be due to prior exposure to red
- form as a result of exposure to environmental sources such as cell antigens from pregnancy or transfusions.
pollen, fungus, and bacteria, which have a structure similar to
some RBC antigens. ● Tentative interpretation, example tentative detection of presence
or absence of antibodies.
● Passively acquired antibodies ● The cells here- I, II, III- these are red cell antigens.
- produced in one individual and then transmitted to another ● Per red cell, there are different antigens present. That's why
individual via plasma-containing blood components or para mas mag detect ang presence ng unexpected antibodies,
derivatives such as intravenous immunoglobulin (IVIG) we do II cell or III cell screen
● PHASES= immediate spin 37°C and then the AHG phase
● Autoantibodies ● Here in check cells, zero that is negative. If 1+, 2+, 3+, and 4+,
- antibodies directed against antigens expressed on one's own positive reaction. Graded based on strength or degree of
RBCS and generally react with all RBCS tested. reactivity.
Antibodies considered significant or termed as clinically significant ● If positive in immediate spin= IgM class
antibodies that causes decrease survival of RBCS possessing the ● If positive at 37°C and positive AHG = IgG class
target antigen. ● Remember: final reading in AHG phase, nandito yung mga
How? clinically significant antibodies

1. There’s reactions AHG so immunoglobulin class is IgG. Single

specificity kasi sa AHG lang may reaction. Alloantibody kasi no
reaction in DAT. Poly is Polyspecific antiglobulin agent.
Alloantibody since no reactions in Polyspecific antiglobulin
reagent
We can identify antibody or auto antibody present by autocontrol
and direct antiglobulin test.
(+) autocontrol and (+) direct antiglobulin test = autoantibody
present
(+) autocontrol and (-) direct anti globulin test = false positive (if
false positive, ulitin ang nasa panel using a different type of ● Reagent red cells also called RBC reagents or reagent RBCS or
potentiator or no enhancement solution) simply screening cells.
● Group O cells are used so that anti A and anti B will not
2. IgG. There are reactions in 37°C and AHG phase, and then interfere in the detection of antibodies to other blood group
multiple specificities kasi sa dalawang cases may positive systems.
reactions and then it's an Alloantibody kasi no reactions at ● purpose of incubation = allow antibody binding.
polyspecific antiglobulin reagent ● Coombs or check cells
● addition of check cells to know if it's true negative, for
confirmation
● positive = agglutination, no need to add check cells
● negative = add check cells

ULITIN:
● if may reaction or agglutination upon additional check cells =
true negative, no antibody detected
● (-) + check cells = (+) result = true negative
● (+) = no need to add check cells

● Advantages
o Flexibility of the test system
o Availability of required testing equipment
o Relative low cost.

● Disadvantages
3. IgM since there’s reaction in immediate spin. Alloantibody kasi o Instability of the reactions and subjective nature of grading
no reactions in DAT by the technologist
4. IgM and auto antibody, observe the reactions o Amount of hands-on time for the technologist
5. IgG and auto antibody. o Problems related to the failure of the washing phase to
remove all unbound antibody
TUBE METHOD
In antibody screen, we perform methods to detect these TUBE METHOD: RBC reagents
clinically significant antibodies from traditional which is the tube to ● The RBC reagents used in the antibody screen come from
automated methods, gel technology, solid phase and affinity column group O individuals who have been typed for the most common
technology. and the most significant RBC antigens
● Usually obtained from commercial manufacturers
● The traditional tube method of detecting antibodies is an ● Cells are suspended at a concentration between 2-5% in a
indirect antiglobulin test performed in a test tube. preservative diluent, which maintains the antigens and prevents
● It may include immediate spin (IS) phase to detect antibodies hemolysis.
reacting at room temperature (but not required and may lead to ● AABB standards require that antibody detection in recipients
the detection of clinically insignificant cold antibodies), usually of be performed using reagent cells that are NOT pooled.
IgM class Pooled= combined
● Must include 37C incubation phase But antibody screening pooled ang reagent that consists of two
to three group or reagent cells selected from different donors of
known phenotype.
● Sets of alleles are prepared so that all antigens to the most
commonly encountered blood group antibodies are presented
and must include C, c, D, E, e, Fya, Fyb, Jka, Jkb, K, k, Le(a),
Le(b), P1, M, N, S, and s antigens.
● A three-cell screening panel ensures homozygosity for most
antigens against which the corresponding antibodies are likely
to exhibit dosage, but the additional cell adds expense to
pretransfusion testing

TUBE METHOD: Enhancement Reagents or Potentiators

COOMB’S CONTROLL CELLS (CHECK CELLS)

● Group O Rh (+) RBCS sensitized with anti IgG.
● It is added to negative AHG tests to validate or to confirm the
negative reaction.

In AHG test, two possible results

● negative result = add check cells to confirm true or negative
reaction.
● Upon addition, if no agglutination = invalid test that is false
negative
● Pwede neutralize AHG reagent, neutralize = blocked antibody
sites. That is due to inadequate red cell washing.
● Purpose of washing is to remove unbound antibodies or
proteins.
● If inadequately wash unbound antibodies or proteins present
parin, sila ang magbind sa AHG reagent so inhibited yung
reaction ng sensitize RBCs, walang visible agglutination
● Another reason maybe expired reagent or forgot to add in
procedure
● Another negative result, if we add check cells and there’s
agglutination = true negative
● The test was done properly and there is no sensitization of
patients antigen or RBC.

GEL TECHNOLOGY or COLUMN AGGLUTINATION

● Alternative method for Alloantibody detection
tube method = most common procedure in antibody screen.
Gel tech same sensitivity in tube testing but more standardized.
Pinaka advantage ng gel technology, standardization. Compared in
tube mas reliable, stable and well defined ang agglutination reaction
TUBE METHOD: AHG Reagents in gel technology
● Monospecific or polyspecific. ● Controlled centrifugation red cells through the
Monospecific dextran-acrylamide gel and appropriate reagents predispense
- either anti IgG or antibodies to complement components in specially designed microtube
C3 and C4 or C3B and C3D Gel particles ideal for trapping ang agglutinates
Poly specific Gel tech used not only in antibody detection and identification but
- both IgG and complement components. also in
- It's not often used when performing antibody detection ● Can do ABO forward and reverse grouping, Rh typing, direct
testing since the presence of anticomplement in the AHD antiglobulin test (DAT), antibody screening, antibody
reagent may lead to the detection of clinically identification and compatibility testing (cross matching)
insignificant antibodies. Rh typing, the gels in the card have Anti A, Anti B , Anti D, A1 and B
cells
Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne c
Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne c
● Principle: hemagglutination.
ABO and RH typing This shows young principle in
gel technology.
Gel card consists of gel
matrix since antibody
detection, anti-IgG is the
antibody in gel. Then we will
add RBCS. If present ang red
gel test of compatibility testing and cell in target antigen it will
antibody detection and bind with antibody, anti IgG
identification. Micro tubes are filled and then it will result in agglutination, that is now your positive
with gel containing anti-IgG as reaction.
shown here. Note: ideal for trapping agglutinates.
positive regardless of grading, distributed sila sa upper part
Reactions in gel test same in absent antigen, wala magbind sa anti IgG, no reaction. Red cell will
grading agglutination in test tube settle at the bottom that is negative reaction.
Hemeagglutination technique
Also graded from one to four Procedure
● Add 50 uL of 0.8% suspension of reagent screening sells to the
In gel tech we use a concentration of red cells that is lower in tube microtubes of the anti-IgG gel cards
method. We use 0.8 percent suspension of red cells, standard ● Add 25 uL of patient serum or plasma to the microtubes
concentration RCS that is 3 to 5%. ● Incubate the gel card at 37C for - 15 minutes -a predetermined
Develop gel technology = Doctor Yves Lapier. time and centrifuge - 10 minutes
MUST READ! ● After centrifugation, the test results are read and graded. No
washing step is required for antiglobulin testing. No
IgG-sensitized red cells are required

purpose of incubation = to allow time to bind antigen antibody

complex.
Larger agglutinates are trapped at the top of the gel microtubes.
Smaller agglutinates travel through the gel microtubes and may
be trapped in either top or bottom half of the microtubes.
Unagglutinated cells settle at the bottom

SOLID PHASE ADHERENCE TEST

Another alternative for all antibody detection
Steps:
a. Reagent RBCS are bound to the bottom of microplate
wells
Serum and an enhancement reagent are added and
incubated at 37° Celsius, allowing the antibody to bind to
AGGLUTIONATION REACTION the solid phase.
b. After washing, indicator RBCS coated with anti-human IgG
or added, and the plates are centrifuge
c. ballet young wells coated with red cell.

● Column agglutination or gel

technology has
equivalent sensitivity to standard LISS tube testing for antibody
detection and is particularly sensitive for detection of anti-Rh
antibodies.

● A gel cord measures 5x7 cm

and consists of 6 microtubes. Each
microtube contains predispense
gel diluents and reagents, if
applicable.

● Major Advantage:
standardization
o Less sensitive to clinically insignificant cold antibodies,
eliminating unnecessary antibody workups o Wells coated with red cell.
● Disadvantage: need special instrument or equipment. o We will add enhancement reagent usually LISS
● Centrifugation time: 10 minutes o Then add young patient serum incubate at
● Incubation time: 15 minutes o 37°C and then wash add, indicator cells coated with anti IgG
and then centrifuge.

Cell number – per red cell, different antigens are present

Patient cells – this is where we can identify if alloantibody or
autoantibody is present

EXCLUSION

● When interpreting panel results, the first step is to

exclude antibodies that could not be responsible for the
Solid phase advantage over tube testing = Endpoint agglutination reactivity seen.
poll is more clearly defined. ○ Cells that gave a NEGATIVE reaction in all
phases of testing are examined
■ Immediate spin, LISS/37C, AHG
○ The antigens on these negatively reacting cells
will probably not be the target of the antibody
(rule-out technique)

EVALUATION OF PANEL RESULTS

● Evaluation of panel results should be carried out in a

logical step-by-step method to ensure proper
Summary of anti-gram Tentative interpretation, Guide for identification and to avoid missing antibody specificities
interpretation. that may be masked by other antibodies

ANTIBODY PANEL IDENTIFICATION

● Antibody identification is accomplished by first testing an

extended panel of reagent RBCs of known phenotype
against the recipient’s serum by IAT technique
● An autocontrol is always included as part of the testing to
help differentiate whether autoantibody, alloantibody or
both are present

PATIENT HISTORY

● Information concerning the patient’s age, sex, race, RULE OF THREE

diagnosis, transfusion and pregnancy history,
medications, and IV solutions may provide valuable ● Confirming the presence of an antibody by
clues in antibody identification studies, particularly in demonstrating three cells that react and three cells that
complex cases do not react
● It’s important to refer to medical records for patient ● The p value must be 0.05 or less for identification to be
history in cases of emergency – we can use these considered valid
records to make conclusions or initial interpretations

● Antibodies may be removed from serum by adding the

target antigen and allowing the antibody to bind to the
antigen (similar to the neutralization technique)
● Antigen/antibody complex is composed of solid
precipitates and is removed from the test system by
centrifugation
● The absorbed serum is tested against an RBC panel

Autoantibodies and Autoimmune Hemolytic Anemia

● The initial investigation of an antibody includes a DAT or

Answer – allo-Fy(a) autocontrol that, if positive, could be caused by a cold
autoantibody, warm autoantibody, drug mechanism,
ADDITIONAL TECHNIQUES: HDFN, or a delayed transfusion reaction.
ENZYMES
Associated with Specificity Serum Antibody
● When it appears that multiple antibodies may be
present in a sample, treatment of the panel cells with Transfusion reaction IgG Specific alloantibody
enzymes may help separate the specificities and allow
for identification. Warm autoimmune disease IgG (C3) Reacts with all cells at
antihuman globulin phase

Cold autoimmune disease; C3 Reacts with all cells at colder

pneumonia temperatures

Drug interaction IgG (C3) Serum may be nonreactive

Clot tube stored at 4° C C3 None

HDFN; maternal antibodies on IgG Alloantibody or ABO antibody

infant’s RBCs from mother on blood cells

● The specificity of cold and warm autoantibodies is

usually not relevant because autoantibodies are reactive
with all reagent red cells, donor red cells, and autologous
cells, regardless of the antigens present

Cold Autoantibodies and Cold AIHA

● Cold AIHA is caused by IgM autoantibodies, which

optimally react at temperatures below 37° C
Two Procedures for Enzyme Treatment: ● Cold AIHA disease is called cold agglutinin disease
1. One-Stage Enzyme Technique: antibody identification (CAD) (I antigen)
technique that requires the addition of the enzyme to the
cell and serum mixture Cold Agglutinin Disease
a. Patient serum, papain, and red cells are ● In CAD, patients may have a history of mild anemia,
incubated together Mycoplasma pneumoniae infection, or infectious
2. Two-Stage Enzyme Technique: treatment of the red cells mononucleosis.
with an enzyme before the addition of the serum ● A chronic form of CAD is often seen in elderly patients
a. Panel or screening cells are pretreated with with associated lymphoma, chronic lymphocytic
enzymes ficin or papain, then washed leukemia, or Waldenström macroglobulinemia
b. The pretreated cells are used without other ● The autoantibody specificity in CAD is most often
enhancement media in the antiglobulin test autoanti-I and less commonly autoanti-i

NEUTRALIZATION Paroxysmal Nocturnal Hemoglobinuria

● Paroxysmal cold hemoglobinuria (PCH) is a rare
autoimmune disorder characterized by hemolysis and
hematuria associated with exposure to cold
● autoanti-P is associated with this immune hemolytic
anemia
● Auto-anti-P is an IgG antibody reacting weakly or not at
all in routine in vitro test methods
● The Donath-Landsteiner test is required for confirmation
of PCH
● A patient with PCH needs to be transfused with an RBC
unit; it should be administered through a blood warmer
and patients should be kept warm at all times.

Dealing with Cold Autoantibodies

● Recognizing the presence of a cold autoantibody is the
first step in beginning the investigation
● Cold autoantibodies can have reactions at several
phases, and each example may be different in the
reaction strength and phase noted

Cold Autoantibody Specificity

● The tool first used is called a cold panel, which is a set of
selected reagent red cells that aids in determining the
presence of a suspected cold autoantibody

RBCs + PRP 2,000 x g, 3 minutes

CONTENTS: APPROVED PRESERVATIVE SOLUTIONS:
● Citrate ● Acid citrate-dextrose (ACD)
● Adenine ● Citrate-phosphate-dextrose (CPD) Harvesting platelets or 5,000 x g, 7 minutes
● Glucose/Dex ● Citrate-phosphate-dextrose-adenine cryoprecipitate from plasma
trose (CPDA-1)
● Phosphate ● Citrate-phosphate-2-dextrose (CP2D)
● Citric acid ● ACD-A (anticoagulant citrate-dextrose Plasma Expressors
A): used in apheresis procedures with ● mechanical devices that apply pressure to the blood bag
a shelf life of 21 days
which allows blood components to flow from one bag to
another by way of the integrated tubing system
Red Cell Additive Solutions
“SAM-G” Scale
● Some components, like plasma, must be labeled within
CONTENTS: ADDITIVE SOLUTIONS:
● Saline ● Adsol (AS-1) 10% of the actual volume of the components.
● Adenine ● Adenine (AS-3) ● Scales should be validated to ensure that they produce
● Glucose ● Optisol (AS-5) accurate and reliable results.
● Mannitol ● SOLX (AS-7)
Tubing sealers
Platelet Additive Solutions ● use a combination of targeted radio frequency energy
● Platelet Additive Solution (PAS): is a buffered salt and pressure to melt and seal the tubing.
solution that replaces a portion of the plasma used to ● The tubing is placed in a channel, and a trigger is
store platelets activated when the user is ready to make the seal.
● PAS platelets are leukocyte-reduced apheresis platelets
that are stored in a mix of 65% PAS and 35% plasma Sterile Connection Device
● PAS were developed to increase platelet viability during ● SCDs allow two separate blood bags to be connected
storage, minimize the amount of plasma in platelet via their PVC tubing without breaching the integrity of

Transport 1-10C Washed RBCs

● Increase RBC mass of symptomatic anemic patients with
Shelf-life Depends on the anticoagulant; if history of allergic, febrile, urticarial, and anaphylactic
exposed to open air – 24 hrs. reaction
● WB is prepared in an aseptic manner with a ratio of 14
mL of anticoagulant preservative solution per 100 mL of Indication • PNH
whole blood • For patient with polyagglutination
• For patient with aplastic anemia
• Patients with IgA-deficiency
Packed RBCs
● Increase RBC mass of symptomatic, normovolemic Storage 1-6C
patients
Shelf-life 24 hrs. (open system)
Indication ● Oncology patients undergoing
chemotherapy or radiation therapy
● Trauma patients
● Patients undergoing cardiac, ● Washing is accomplished with approximately 1000 to
orthopedic, and other surgeries 2000 mL of 0.9% saline using the automatic blood cell
● Patients with end-stage renal disease processor described for deglycerolizing frozen RBCs.
● Premature infants and neonates
● Patients with sickle cell disease
Frozen RBCs
Storage 1-6C ● For long-term preservation to maintain an inventory of
rare units or to extend the availability of autologous units
Shelf-life Depends on the anticoagulant

Indication For rare blood and autologous units

Leukocyte-Reduced RBCs
● Increase RBC mass in patients with severe and or Storage -65- -120C
recurrent febrile transfusion reactions due to leukocyte
Shelf-life 10 years
antibodies

Storage 1-6C Indication Febrile reactions

Shelf-life 24 hrs (open system), if Storage Room temperature

automated machines
(closed system) – 14 days
Shelf-life 5 days

DEGLYCEROLIZATION PROCESS:
Fresh Frozen Plasma & Plasma Frozen within 24 Hours of
● Frozen RBCs are thawed in a 37C dry warmer or water
Phlebotomy
bath.
● FFP: contains all the coagulation factors, including labile
● Unit is washed in a series of saline solutions of
factors V and VIII, which do not store well at
decreasing osmolarity.
temperatures greater than -18C
● A visual check of the supernatant from the final wash is
● PF24: may have reduced levels of factors V and VIII
performed to ensure sufficient glycerol removal and
compared with FFP
minimal hemolysis

Irradiated RBCs Indication • Management of bleeding in patients who require coagulation

factors II, V, X, or XI, when the concentrates are not available or
● Gamma irradiation of cellular blood components are not appropriate
prevents proliferation (multiplication) of the donor’s T • Abnormal coagulation assays resulting from massive transfusion
lymphocytes that cause transfusion-associated GVHD • Management of patients anticoagulated with warfarin who are
bleeding or require emergency surgery
• Replacement solution for therapeutic plasmapheresis for the
Indication • Prevent TA-GVHD treatment of TTP and HUS (plasma cryoprecipitate reduced can
• Immunocompromised patient/immunodeficiency also be used for these patients)
• Intrauterine/exchange transfusion • Correction or prevention of bleeding complications in patients
• Relative/family-related who have severe liver disease with multiple factor deficiencies
• HLA-matched donor • Management of patients with DIC when the fibrinogen level is
• Patient with cancer less than 100 mg/dL
• Patient who went chemotherapy • Management of patients with rare specific plasma protein
• BM stem cell transplantation deficiencies

Storage 1-6C Storage -18 to -65C

Shelf-life 28 days or end of the expiration date of the unit Shelf-life -18C (1 year)
-65C (7 years)

● Current FDA guidance suggests a midplane dose of 25

Gy (2500 cGy/rad), with a minimum dose of 15 Gy to any Cryoprecipitated AHF
point of the blood container ● Also referred to as cryoprecipitate or CRYO – cold
insoluble precipitate that forms when a unit of FFP is
Platelet Components thawed between 1-6C
● To control or prevent bleeding associated with critically ● Contains, in a concentrated form, most of the
decreased circulating platelet numbers or functionally coagulation factors that are found in FFP (vWF,
abnormal platelets fibrinogen, FVIII, fibronectin, FXII)

Indication • Cancer patients undergoing chemotherapy or Indication • Fibrinogen deficiency

radiation • Hemophilia A
• Recipients of hematopoietic progenitor cell • VWD
transplants for a period after transplant • Factor XIII deficiency
• Patients with postoperative bleeding
• Organ transplant patients Storage -18C

Storage 20-24C with agitation Shelf-life 1 year

Shelf-life 5 days, midnight of 5th day

CONDITION SHELF-LIFE

POOLED PLATELETS If stored at -18C 1 year

● To achieve a therapeutic dose, platelet concentrates are
pooled for transfusion in adults If thawed 6 hours

If pooled 4 hours
Indication • Thrombocytopenia
• DIC
• Bleeding If thawed and pooled 4 hours

Storage Room temperature

● QC of CRYO must demonstrate greater than 150 mg of
Shelf-life 4 hrs
fibrinogen and 80 international units of factor VIII per unit
tested

Transportation of Blood Components

Plasma Protein Fraction Albumin ● Transport Temperature: 1-10C (Whole Blood and RBCs)
● Frozen units are shipped on dry ice.
Indication Replace loss of colloids in
● Platelets must be maintained as close as possible to
hypovolemic shock, severe 20-24C during shipping.
burns, or for pressure support ○ Discontinuation of the agitation of platelets
during hypotensive episodes during transportation should not exceed 24
hours.
Storage 2-10C
● On receipt of a shipment of blood components, the
temperature and appearance of the units must be
Shelf-life 5 years
observed and recorded.

Checklist for Receiving Blood

DISTRIBUTION AND ADMINISTRATION
✓ Temperature acceptable for component
Labeling
✓ Appearance: clots, discoloration,
1. Donation Identification Number
hemolysis
2. ABO/Rh Blood Groups
✓ Container closure
3. Collection Date (optional)
✓ Attached segments intact: RBCs
4. Produce Code
✓ Expiration date and time
5. Expiration date (and time)
✓ Shipping list correct
6. Special Testing (optional)
✓ Intact labels

Administration of Blood Components

Requirements of safe blood administration include the following:

● Positive identification of the patient using two
independent identifiers before transfusion is critical to
avoid transfusion reactions that may be fatal.
● A system to avoid and detect clerical errors also
contributes to avoiding serious reactions. Strict
adherence to policies regarding identification numbers
and mislabeled tubes should be followed.
● Direct observation of the patient should occur during the
first 15 minutes after infusion begins and periodically

Fever – most common symptom

Pain at infusion site
DIC

Clinical Signs Indicative of a Transfusion Reaction:

● Fever ≥1C increase or >38C
● Chills and rigors
● Respiratory distress: wheezing, coughing, dyspnea,
cyanosis
● Hypertension or hypotension
● Pain: abdominal, chest, flank or back, infusion site
● Skin manifestations: urticaria, rash, flushing, edema
● Jaundice or hemoglobinuria
● Nausea or vomiting
● Abnormal bleeding
● Oliguria or anuria Saline – to maintain blood pressure
Four Leading Causes of Preventable Laboratory Errors:
1. Improper specimen identification
2. Improper patient identification
3. Antibody identification error
4. Crossmatch procedure error

HEMOLYTIC TRANSFUSION REACTION (HTR)

● Immediate: at time of transfusion

● Delayed: few (3 to 7) days after transfusion
● HTRs most often occur with the transfusion of
incompatible RBCs but may also occur with transfusion
of ABO-incompatible plasma containing products.
○ Wrong blood type is transfused into the patient
Delayed Hemolytic Transfusion Reaction (DHTR)
○ Can be immediate or acute; during transfusion
● Appears after 24 hrs
or a few days post transfusion, the patient has a
● Most often a result of an anamnestic response
reaction
(secondary response) in a patient who has previously
been sensitized by transfusion, pregnancy, or transplant
Immediate Hemolytic Transfusion Reaction (IHTR)
and in whom antibody is not detectable by standard
● Underlying Cause: transfusion of an incompatible whole
pretransfusion methods
blood or RBC product to a recipient
○ It is possible that in the recent blood transfusion
● Four Most Commonly Identified RBC Antibodies:
of the patient, they were already exposed to the
○ Anti-A
foreign antigens of the donor and developed
○ Anti-Kell
antibodies

Febrile Nonhemolytic Transfusion Reaction

● “a 1C temperature rise associated with transfusion and
having no medical explanation other than blood
component transfusion”
● Any 1C or greater temperature increase above the
patient’s baseline temperature, during or within 24 h after
transfusion, with a minimum recorded temperature of
38C
● A 1C temperature increase above the patient’s baseline
pretransfusion temperature during or within 8h after the
end of the transfusion
● FNHTR is the most common encountered type of
transfusion reaction
○ Most common cause if cytokines produced by
leukocyte antibodies
○ Resolution – pre medication with
acetaminophen for reducing temperature, or in
Fever – most common sign of HTR cases wherein we need to transfuse blood, we
use leukoreduced units
Acute vs Delayed HTRs
Clinical Signs & • Fever: temperature increase ≥1C (or 2F) above 37C
Acute Hemolytic Delayed Hemolytic Symptoms • Other symptoms: chills, rigors, headache and vomiting

Clinical Signs • Immediate or within 24h • >24h to 28 days Major • Nonthreatening

& posttransfusion posttransfusion Complications • Significant discomfort to the recipient
Symptoms • Fever, chills, flushing, pain • Fever: temperature
at site of infusion, increase ≥1C (or 2F) with or Causes • Antibody to donor WBCs
tachycardia, tachypnea, without chills • Cytokines released by WBCs during blood product
lower back pain • Unexplainable decrease storage
• Hemoglobinemia, in hemoglobin and
hemoglobinuria, hypotension hematocrit
Clinical • DAT: negative
• Jaundice and
Laboratory Tests • No visible hemolysis
hemoglobinuria

Management • Antipyretics: acetaminophen

Allergic Transfusion Reactions

Prevention • Prestorage leukocyte reduction of blood products Urticarial Anaphylactic

Clinical Signs & • Hives and itching within • Rapid onset and severe
Allergic (Urticarial) Transfusion Reactions Symptoms 15-20 minutes of wheezing, coughing,
● In the classic type I hypersensitivity response, preformed transfusion dyspnea, bronchospasm,
IgE antibodies in the recipient react with the allergen respiratory distress,
vascular instability
(plasma protein), which activates mast cells.
● Mast cell activation results in degranulation and the
Major None • Shock
release of histamine, proteases, and chemotactic Complications • Loss of consciousness
factors. • Mortality
○ Histamine – primary mediator of allergic
response; released when antigen binds to the Causes • Recipient antibodies to • Associated with genetic
foreign plasma proteins IgA deficiency in recipient
surface of tissue mast cells or other substances such who possesses IgG
SIGNS & SYMPTOMS as drugs or food complement-binding anti-IgA
● Local erythema (redness) consumed by blood donor antibodies
● Pruritus (itching)
● Hives (raised, firm, red welts) Clinical • DAT: negative • DAT: negative
Laboratory • No visible hemolysis • No visible hemolysis
PREVENTION Tests • Perform IgA antigen and
● Medication and premedication with antihistamine anti-IgA testing
● Transfusion of plasma-deficient blood components
Management • Transfusion interrupted • Transfusion terminated
Anaphylactic Response • Antihistamine • Epinephrine and similar
administered drugs administered
● Immediate hypersensitivity type of immune system • Oxygen administered and
response open airways maintained
○ Anaphylactic and allergic responses are of both
immediate hypersensitivity type, but Prevention • Premedication with • Plasma-containing products
anaphylactic is more severe antihistamine, if patient from IgAdeficient donors
history reveals repetitive • Washed red cell and
○ Allergic – single organ reactions platelet products
○ Anaphylactic – involves multi organ • May necessitate washed
● Two significant features distinguish anaphylactic cellular products
response from other types of transfusion reactions:
1. Fever is absent
Transfusion-Related Acute Lung Injury
2. Clinical signs and symptoms occur after
●
Noncardiogenic pulmonary edema
transfusion of just a few milliliters of plasma or ●
Currently the leading cause of mortality from transfusion
plasma-containing blood components
○ Leading cause of transfusion-associated
● Anaphylactic and anaphylactoid reactions are the most
fatalities surpassing ABO incompatibility and
fatal and deadly as transfusion reactions
bacterial contamination
● Attributed to IgA deficiency in patients who have ● Caused by anti-leukocyte antibodies; anti-HLA class I;
developed anti-IgA antibodies by sensitization from
anti-neutrophil antigens that are present in plasma of the
transfusion or pregnancy
transfused unit
○ Selective IgA deficiency – most common
● Blood component is leukoreduced or leukopoor
immunodeficiency; rare ● TRALI – involvement of lungs; much more dangerous
○ Either low levels of IgA or no IgA
compared to FNHTR
○ <5 mg/mL – severe
SYMPTOMS:
○ >5 mg/mL – partial deficiency
● Dyspnea
SIGNS & SYMPTOMS: ● Hypoxemia
1. Anaphylactic, in patients deficient in IgA who have
● Tachycardia
class-specific IgA antibodies
● Fever
2. Anaphylactoid, in patients having normal levels of IgA
● Hypotension
but a limited type-specific anti-IgA that reacts with light ● Cyanosis
chain (kappa or lambda) of donor’s IgA
● Lack of
THERAPY & PREVENTION:
abnormal
● Stop the transfusion, and do not restart transfusion of the
breath sounds?
blood component. ● X-ray shows pulmonary edema
● Keep the IV line open with normal saline.
● Give epinephrine (usually about 0.5 mL of 1:1000
solution) immediately. Clinical Signs & • Fever (1-2C increase)
Symptoms • Chills
● For severe reactions, corticosteroids or aminophylline, or • Hypoxia
both, may be indicated. Airway patency must be • Dyspnea (shortness of breath)
maintained, and the vital signs must be stabilized by • Cyanosis (bluish discoloration of skin)
appropriate means. • Nonproductive cough
• New onset bilateral pulmonary edema
• Hypotension
• Acute onset within 6h of blood transfusion

Major • Severe and dramatic presentation

Complications • Can be fatal

Causes • Interaction of recipient-related risks and transfusion event

contamination of blood components include:
• Donor with pregnancy history, anti-HLA, anti-HNA ○ Closed sterile system of collection bags for use
in collection and blood product manufacturing
Clinical • DAT: negative ○ Attention during the donor phlebotomy process
Laboratory • No visible hemolysis to scrub arm at and around the venipuncture
Tests • WBC antibody screen in donor and recipient
• Chest x-ray
site
○ Diversion of the skin plug, generated during
Management Respiratory support phlebotomy, into a sample diversion pouch
○ Donor health check before donation
Prevention Avoid use of plasma components from multiparous women ○ Post Donation information provided to donors to
who have HLA antibodies report any health issues after donation
○ Requirement for methods to limit and to detect
Transfusion-Associated Circulatory Overload (TACO) or inactivate bacteria in all platelet components
● Associated with the lungs, but the main is the heart
● Second most common cause of transfusion related Yersinia According to CDC, it is the MOST COMMON CAUSE OF
deaths reported to the FDA enterocolitica DEATH by bacterial contaminated blood components
● Iatrogenic (physician-caused) transfusion reaction
Pseudomonas Second most common isolate found in RBC units
● Most frequent cause: transfusion of a unit at a too fast a spp. P. putida, P. fluorescens
rate – 100 mL/hr (usual is 200 mL/hr)
SIGNS & SYMPTOMS: Propionibacterium Common isolate of human skin; was the most common
● Elevation of B-type Natriuretic Peptide – secreted from acnes bacterial contaminants in RBC
the ventricles of the heart
● Dyspnea, orthopnea, cyanosis, tachycardia, elevated Bacillus cereus, Most frequently recovered from donated blood and
blood pressure, pulmonary edema, jugular venous Staphylococcus contamination of platelet
epidermidis *S. epidermidis or S. aureus are the MOST COMMON
distention, pedal edema and headache BACTERIAL CONTAMINANTS of blood (Harmening, 7th
INTERVENTION: According to CDC: edition)
● Stop transfusion or infusion of other fluids. Y. enterocolitica *S. aureus accounts for the greatest number of
deaths due to blood product contamination reported to
● Stabilize patients. According to FDA: the FDA from 2011 to 2015, all of which were platelet
● Contact the physician and notify the blood bank. S. aureus products.
● Provide respiratory support (eg, supplemental oxygen,
ventilatory assistance). NOTE:
● Administer diuretics were not contraindicated. • Brown plasma, purple plasma, clots and hemolysis = blood unit should not be
used for transfusion because they are signs of bacterial contamination
● Perform blood bank investigations to rule out other • Green color plasma = usually associated with biliverdin; discard
adverse effects • Platelet concentrate is the most common blood component associated with
transfusion sepsis.

Clinical Signs & • Acute respiratory distress (dyspnea, cough)

Symptoms • ↑ BNP PREVENTION AND TREATMENT:
• ↑ Central venous pressure
• Left heart failure ● Administer IV antibiotics
• Positive fluid balance ● Carefully inspect blood products before distribution for
• Radiographic evidence of pulmonary edema transfusion
• Acute onset within 6h of blood transfusion
● Proper sterilization technique
● Visual inspection of units (check for clots, and changes
Major Acute pulmonary edema
Complications of color of plasma, or hemolysis)
● Phlebotomy diversion consists of collecting the first
Causes • Transfusion of large volumes of blood component and 20-30 mL of blood in a separate container to be used for
fluids testing
• High flow rates

DELAYED NONHEMOLYTIC TRANSFUSION REACTION

Clinical Laboratory • Rule out TRALI
Tests • Chest x-ray
Alloimmunization
Management • Respiratory support ● May result from prior exposure to donor blood
• Administer diuretics when possible components
● Clinical signs and symptoms may be mild, including
Prevention • Infusions of large volumes of plasma avoided slight fever and falling hemoglobin and hematocrit levels;
• Transfuse only RBC units
• Transfuse blood at slower rate or severe, including platelet refractoriness with bleeding
● Significant complication: even very small amounts of
donor antigenic RBCs can elicit an alloimmune
Bacterial Contamination Reactions
● Caused by psychrophilic bacteria – cold loving Post-Transfusion Purpura
● Major causes: ●A rare complication, usually involving platelet
○ Donor has asymptomatic bacteremia at time of concentrates
donation ● Characterized by a rapid onset of thrombocytopenia as a
○ Rare bacterial infection that can survive in result of anamnestic production of platelet alloantibody
storage conditions recommended for RBCs and ● Usually occurs in multiparous females
platelet components SIGNS & SYMPTOMS:
○ System failure: pinhole in blood collection set, ● Purpura
testing error, not following SOP ● Thrombocytopenia
● Current mitigations in place to assist in the ● Hematuria

Indications for Irradiation of Blood Components for Prevention of TA-GVHD

Absolute Indications • Congenital cellular immunodeficiency

• Hematopoietic progenitor cell transplantation
• Hodgkin lymphoma
• Granulocyte transfusions
• Intrauterine transfusions
• Transfusion to neonates who have received IUT
• Transfusions from biological relatives
• Chemotherapy with purine analogs (fludarabine)

Probable Indications • Low-birthweight infants (<1,200 g)

• Hematologic malignancies other than Hodgkin
lymphoma
• HLA-matched platelet concentrates TRANSFUSION REACTION INVESTIGATION
• High-dose chemotherapy, radiation therapy, and/or
aggressive immunotherapy
DURING TRANSFUSION REACTIONS
● Stop the transfusion
Controversial • Solid organ transplantation
Indications • Large-volume or exchange transfusion of infants who ● Keep IV open with saline
did not receive IUT ● Perform clerical check for ID errors
• Aplastic anemia ● Contact treating physician
• Absolute lymphopenia
● Monitor/record vital signs
Irradiation NOT • HIV infection
Indicated • Hemophilia
• Small-volume transfusions of term infants who did not
receive IUT
• Elderly patients
• Typical dose immunosuppressive therapy (other than
purine analogs)
• Immunocompetent surgical patients
• Pregnancy
• Red cell membrane, metabolic, or hemoglobin
disorders

Iron Overload
● Transfusion-induced hemosiderosis
● Results from the accumulation of excess iron in WORK UP – TRANSFUSION INVESTIGATION
macrophages in various tissues ● Examination of pretransfusion clotted blood specimen,
● Iron intake (250 mg/unit) exceeds the daily iron excretion an EDTA anticoagulated posttransfusion blood specimen
(1 mg/day) with the subsequent deposition of excess iron (perform DAT), and the blood bag
in the liver, heart, kidney ● Perform gram stain on the blood in the bag and culture
● Patients who are transfusion-dependent are at risk of ● Repeat the ABO/Rh typing, antibody screen, and
iron overload (sickle cell anemia); other s at rick are crossmatching
people with hemoglobinopathies and thalassemia ● Examination of posttransfusion urine

Any investigation of a suspected transfusion reaction must include

investigation of clinical data to include the following:
1. Diagnosis
2. Medical history of pregnancies, transplants, and previous
transfusions
3. Current medications
4. Clinical signs and symptoms of the reaction

Depending on the preliminary investigation results, more

specimens may be required:
1. A clotted blood specimen drawn 5-7 hours after
transfusion for unconjugated indirect bilirubin ○ Intravascular intrauterine transfusion – The
determination. fetus will be transfused with blood through the
2. The first voided post-transfusion urine collection. umbilical vein and the placenta. This is done
3. Other specimens collected at various times that are when the fetus has low RBC count or anemic
considered appropriate to the transfusion reaction ● There are several factors that affect immunization and
investigation. severity of HDFN, including antigenic exposure, host
factors, immunoglobulin class, antibody specificity, and
HEMOLYTIC DISEASE OF THE NEWBORN influence of the ABO group

● Clinical condition caused by the immune destruction of PATHOGENESIS

fetal or neonatal RBCs due to attachment of maternal
antibody (IgG) to an antigen present in the membrane of
RBCs
● The mother can be stimulated to form the antibodies by
previous pregnancy or transfusion and sometimes during
the second and third trimester of pregnancy
● Rh BGS is considered as the most severe cause of
HDFN
○ ABO BGS is the most common cause of HDN
● Rh BGS was considered as the most common cause of
HDFN but the incidence of the disease caused by anti-D
has steadily decreased since 1968 with the introduction
of Rh-immunoglobulin (RhIg)
○ Rhogam
● In addition to the use of RhIg, many other advances
have been made in the diagnosis and treatment of
HDFN
○ Ultrasonography, Doppler assessment of
middle cerebral artery peak systolic velocity,
cordocentesis, allele-specific gene amplification
studies on fetal cells in amniotic fluid, fetal DNA
analysis in maternal plasma, and intravascular
intrauterine transfusion have greatly increased
the success of accurately diagnosing and
adequately treating his disease

BLOOD SYSTEMS CAUSING HDN

● Anti-D (most immunogenic), either alone or in

combination with anti-C or anti-E
● Other Rh antibodies, such as anti-c
● Other blood groups: anti-Kell, anti-Duffy, anto-Kidd
● ABO antibodies (especially from blood type O mother)

○ Liley graph – Zone 1: Mild disease; Zone 2:

Intermediate disease; Zone 3: Hemolytic

Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne c
Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne c
alloimmunization has been determined experimentally
and is known to be less than that required to saturate all
D antigen sites
● FULL DOSE:
○ Contains 3o ul of anti-D
○ Protects up to 30 mL of D+ whole blood
○ After 12 weeks gestation, a full dose (300ug) is
indicated for abortion or miscarriage in
D-negative women
○ If mother is exposed to ≥ 2.5 mL Rh positive red
cells of the baby
○ If full dose vial is not available, micro dose vials
are not allowed
● MICRO DOSE
○ Contains 50 uL of anti-D
● Of the non-Rh system antibodies, anti-Kell is considered ○ Protects up to 5 mL of D+ whole blood
the most clinically significant in its ability to cause HDFN ○ After 12 weeks gestation, a 50 ug is indication
● Kell antigens are present on immature erythroid cells in for D-negative females for abortion or
the bone marrow, so severe anemia occurs not only by miscarriage, or ectopic pregnancies
destruction of circulating RBCs, but also by precursors ○ If mother is exposed to ≤ 2.5 mL Rh positive red
cells of the baby
RH HDFN ○ If microdose vial is not available, full dose vial
can be administered
● In Rh(D) HDFN, the Rh positive firstborn infant of a Rh ● Massive fetomaternal hemorrhages of more than 30 mL
negative mother usually is unaffected because the of full blood can occur in less than 1% of deliveries and
mother has not yet been immunized these massive hemorrhages can lead to immunization if
● When the D antigen is inherited from the father, these adequate RH immunoglobulin is not administered.
fetal cells immunize the mother and stimulate the ○ How do we know if there is massive
production of anti-D. Once the mother is immunized to D fetomaternal hemorrhage? We collect maternal
antigen, all subsequent offspring who inherit the D sample which is obtained within 1 hour of
antigen will be affected delivery, then we can do screening tests such
○ Can also be stated as Rh positive father and Rh as Rosette technique
negative mother ○ Rosette technique/test detects Rh fetal positive
● As little as 1 mL of fetal RBCs can immunize the mother red cells in maternal circulation
○ If HDN is positive for screening test, we do
RHOGAM or RhIg confirmatory test; we proceed to the
● Rh Immune globulin (RhIg) is as solution of concentrated quantitation of the hemorrhage by
anti-Rho(D) KLEIHAUER-BETKE TEST
● Prepared from pooled human plasma of patients who
have been hyperimmunized and contains predominantly KLEIHAUER-BETKE TEST (ACID ELUTION TEST)
IgG anti-D
● Inactivate fetal Rh antigens to prevent sensitization from ● Developed by Enno Kleihauer and Klaus Betke
the mother ● Or flow cytometry is used to quantitate the number of
● If the mother is sensitized and is injected with Rhogam fetal Rh-positive cells in the mother’s circulation as a
after delivery, it is ineffective because there are anti-Rh result of a fetomaternal hemorrhage
𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑓𝑒𝑡𝑜𝑚𝑎𝑡𝑒𝑟𝑛𝑎𝑙 ℎ𝑒𝑚𝑜𝑟𝑟ℎ𝑎𝑔𝑒 (𝑚𝐿)
antibodies that are already formed; purpose of Rhogam ● 30
is to prevent sensitization from the mother, but when ○ 30 – amount of fetal blood appropriate to one
sensitization has already occurred, Rhogam is ineffective vial of RhIg
● Rhogam can be administered during pregnancy, before ○ Volume of FMH = % fetal cells x 50
delivery, or immediately after delivery, miscarriage, or ○ Alternative formula for volume of FMH:
abortion # 𝑜𝑓 𝑓𝑒𝑡𝑎𝑙 𝑐𝑒𝑙𝑙𝑠 × 𝑚𝑎𝑡𝑒𝑟𝑛𝑎𝑙 𝑏𝑙𝑜𝑜𝑑 𝑣𝑜𝑙𝑢𝑚𝑒
○ Should be administered within 72 hours of 𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑚𝑎𝑡𝑒𝑟𝑛𝑎𝑙 𝑐𝑒𝑙𝑙𝑠
suspected or proven exposure to Rh positive ● A maternal blood smear is treated with acid and then
red cells stained with counterstain (Shepard’s stain)
● RhIg has two primary uses: ● Fetal cells contain hemoglobin, which is resistant to acid
○ Treatment of ITP and will remain pink/red (deeply stained)
○ Prevention of Rh HDN ● The maternal cells will appear as ghost cells
● Principle: active immunization induced by RBC antigen ● After 2,000 cells are counted, the percentage of fetal
can be prevented by the concurrent administration of the cells is determined, and the volume of fetal hemorrhage
corresponding RBC antibody is calculated
MECHANISM OF ACTION
● The administered RhIg attached to the fetal Rh positive
RBCs in the maternal circulation
● The antibody-coated RBCs are removed by the
macrophages in the maternal spleen
● The RBC antigens are thus unavailable for dendritic cells
to present antigen to T helper cells
● The amount of antibody necessary to prevent

ABO HDFN

● ABO incompatibility between the mother and the

newborn infant can cause HDFN, maternal ABO
antibodies that are IgG can cross the placenta and
attach to the ABO-incompatible antigens of the fetal
RBCs
● Example is when a mother blood type O and the
fetus/newborn is either blood type A, B, or AB
● Destruction of fetal RBCs leading to severe anemia is
extremely rare
● The disease is manifested by the onset of
hyperbilirubinemia and jaundice within 12 to 48 hours of
birth. The increasing levels of bilirubin can be treated
with phototherapy
● As the incidence of HDFN caused by rh(D) has declined,
ABO incompatibility has become the most common
cause of HDFN
● Microspherocytes and increased RBC fragility in the
infant are characteristic of ABO HDFN, but not of Rh
HDFN

DIAGNOSIS AND MANAGEMENT

ABO, RH, and ANTIBODY SCREEN OF THE MOTHER

● The prenatal specimen must be types for ABO and Rh.
the antibody screening method must be able to detect
clinically significant IgG alloantibodies that are reactive
at 37C and in the antiglobulin phase
MCA-PSV
● The measurement of the fetal middle cerebral artery
peak systolic velocity with color Doppler ultrasonography
can reliably predict anemia in the fetus
● MCA-PSV is non-invasive and poses no adverse effects
for the fetus

Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne collado Qatreanne c

Immghy1 Prelims Notes 2
No ratings yet
Immghy1 Prelims Notes 2
43 pages
Intro To Blood Banking
No ratings yet
Intro To Blood Banking
183 pages
Trans BB Lesson 1 Lec
No ratings yet
Trans BB Lesson 1 Lec
8 pages
Lec Transfusion Medicine
No ratings yet
Lec Transfusion Medicine
5 pages
History of Blood Transfusion
No ratings yet
History of Blood Transfusion
3 pages
Reviewer Evaluation Exam
No ratings yet
Reviewer Evaluation Exam
19 pages
L1 Immunohema
No ratings yet
L1 Immunohema
7 pages
BB Day 1 Mtap2 Trans
No ratings yet
BB Day 1 Mtap2 Trans
25 pages
Imhm Prelim Lec
No ratings yet
Imhm Prelim Lec
15 pages
History of Transfusion Medicine Rukmini
No ratings yet
History of Transfusion Medicine Rukmini
103 pages
Blood Transfusion Complication of Blood Transfusion
100% (2)
Blood Transfusion Complication of Blood Transfusion
42 pages
Introduction of Blood Banking
No ratings yet
Introduction of Blood Banking
47 pages
Bloodbanking
No ratings yet
Bloodbanking
7 pages
Blood Product
0% (1)
Blood Product
89 pages
The Blood Donor
No ratings yet
The Blood Donor
14 pages
BB - Introduction To Blood Banking and Transfusion Practices
No ratings yet
BB - Introduction To Blood Banking and Transfusion Practices
6 pages
Blood Banking Lec (Part 1)
No ratings yet
Blood Banking Lec (Part 1)
4 pages
CP Blood Banking - PPTX STUDENT
No ratings yet
CP Blood Banking - PPTX STUDENT
103 pages
Immunohematology
No ratings yet
Immunohematology
195 pages
Immunohematology ASCPi Exam Guide
No ratings yet
Immunohematology ASCPi Exam Guide
22 pages
BB - Lec - 01 - Introduction and History of Blood Banking and Transfusion
No ratings yet
BB - Lec - 01 - Introduction and History of Blood Banking and Transfusion
9 pages
RMT 2 Bloodbank Lecture NOTES
No ratings yet
RMT 2 Bloodbank Lecture NOTES
98 pages
Blood Banking Lab
No ratings yet
Blood Banking Lab
16 pages
Immunohematology for Med Students
No ratings yet
Immunohematology for Med Students
30 pages
Blood Bank
No ratings yet
Blood Bank
43 pages
1 Ih
No ratings yet
1 Ih
29 pages
Medical Technology Donor Screening
100% (1)
Medical Technology Donor Screening
6 pages
Blood Components and Transfusion Guide
100% (1)
Blood Components and Transfusion Guide
36 pages
Introduction To BB
No ratings yet
Introduction To BB
58 pages
Fundamental Concepts: Phases of Transfusion
No ratings yet
Fundamental Concepts: Phases of Transfusion
35 pages
Lec 1 - Intro To Blood Bank
No ratings yet
Lec 1 - Intro To Blood Bank
2 pages
1 Intro To Blood Banking
No ratings yet
1 Intro To Blood Banking
1 page
MLS 046 P1W1
No ratings yet
MLS 046 P1W1
94 pages
A. History of Blood Transfusion: The 1600 - 1700s
No ratings yet
A. History of Blood Transfusion: The 1600 - 1700s
20 pages
BB Notes
No ratings yet
BB Notes
40 pages
Blood Transfusion
No ratings yet
Blood Transfusion
52 pages
Donor Screening and Blood Collection
No ratings yet
Donor Screening and Blood Collection
43 pages
FINALS
No ratings yet
FINALS
23 pages
Blood and Blood Components-Lecture
No ratings yet
Blood and Blood Components-Lecture
13 pages
MBBS Intro To Transfusion
No ratings yet
MBBS Intro To Transfusion
65 pages
Blood Transfusion: Being Presented by
No ratings yet
Blood Transfusion: Being Presented by
55 pages
MTLE BBN
No ratings yet
MTLE BBN
26 pages
8 - Blood Banking and Serology
80% (5)
8 - Blood Banking and Serology
4 pages
8 Blood Banking and Serology PDF
No ratings yet
8 Blood Banking and Serology PDF
4 pages
Donor Selection and Blood Components
No ratings yet
Donor Selection and Blood Components
17 pages
DONOR Blood and Blood Transfusion
No ratings yet
DONOR Blood and Blood Transfusion
16 pages
2-Transfusion of Blood and Blood Products
No ratings yet
2-Transfusion of Blood and Blood Products
47 pages
Mtle Blood Banking Revised
No ratings yet
Mtle Blood Banking Revised
26 pages
Blood 1st Year
No ratings yet
Blood 1st Year
81 pages
Blood Bank
No ratings yet
Blood Bank
26 pages
l6. Blood Donation (Bblab)
No ratings yet
l6. Blood Donation (Bblab)
5 pages
Transfusion Medicine - Yasin Arafat
No ratings yet
Transfusion Medicine - Yasin Arafat
12 pages
Peptalk Proto Donation Proc
No ratings yet
Peptalk Proto Donation Proc
61 pages
Blood Donation
No ratings yet
Blood Donation
16 pages
Donation Lecture
No ratings yet
Donation Lecture
38 pages
Ponciano IH PDF
No ratings yet
Ponciano IH PDF
69 pages
Blood Group Serology
No ratings yet
Blood Group Serology
19 pages
Blood Transfusion Course
No ratings yet
Blood Transfusion Course
42 pages
Gian
No ratings yet
Gian
15 pages
Precipitation - Finals Answers
No ratings yet
Precipitation - Finals Answers
2 pages
Miggy CC2
No ratings yet
Miggy CC2
1 page
CC 2 Miggy
No ratings yet
CC 2 Miggy
1 page
Labeled Immuno Assays Finals Sagot Ni Yoko
No ratings yet
Labeled Immuno Assays Finals Sagot Ni Yoko
3 pages
Blitz CC2
No ratings yet
Blitz CC2
1 page
His Midterm Lesson 1
No ratings yet
His Midterm Lesson 1
2 pages
Curriculum Vitae Pascah
No ratings yet
Curriculum Vitae Pascah
4 pages
Foreign Body Airway Obstruction: (FBAO)
No ratings yet
Foreign Body Airway Obstruction: (FBAO)
14 pages
Cutaneous Manifestations of HIV Infection
No ratings yet
Cutaneous Manifestations of HIV Infection
18 pages
Yoga Guide for Grapplers Fitness
No ratings yet
Yoga Guide for Grapplers Fitness
2 pages
Beating Lyme - 1st Edition ISBN 081440944X, 9780814409442 Open Access Download
No ratings yet
Beating Lyme - 1st Edition ISBN 081440944X, 9780814409442 Open Access Download
17 pages
Elderly Hip Fracture Anesthesia Guide
No ratings yet
Elderly Hip Fracture Anesthesia Guide
3 pages
Neonatal Sepsis
No ratings yet
Neonatal Sepsis
17 pages
Hivduringpregnancy Cbrochure en
No ratings yet
Hivduringpregnancy Cbrochure en
9 pages
Goldberg 2013
No ratings yet
Goldberg 2013
7 pages
Adult Attention-Deficit Hyperactivity Disorder Key Conceptual Issues
100% (1)
Adult Attention-Deficit Hyperactivity Disorder Key Conceptual Issues
12 pages
Handbook of Electrogastrography Instant Reading Access
100% (9)
Handbook of Electrogastrography Instant Reading Access
16 pages
645-Article Text-3548-1-10-20211216
No ratings yet
645-Article Text-3548-1-10-20211216
7 pages
Hormone Cure Instructor Manual PDF
89% (9)
Hormone Cure Instructor Manual PDF
26 pages
روح ٤
No ratings yet
روح ٤
11 pages
Lista de Precios 20-01-2021 (Clientes) - 1
No ratings yet
Lista de Precios 20-01-2021 (Clientes) - 1
32 pages
Reticulocyte Count Lab Guide
No ratings yet
Reticulocyte Count Lab Guide
3 pages
Chapter60 Assessment of Neurologic Function
No ratings yet
Chapter60 Assessment of Neurologic Function
25 pages
Diet Therapy II Mercy Alfred
No ratings yet
Diet Therapy II Mercy Alfred
139 pages
Advanced Radiation Therapy Techniques
No ratings yet
Advanced Radiation Therapy Techniques
18 pages
DRPs in Type II Diabetes Patients at Konawe Hospital
No ratings yet
DRPs in Type II Diabetes Patients at Konawe Hospital
19 pages
1997 Kintzer & Peterson - Primary and Secondary Canine Hypoadrenocorticism
No ratings yet
1997 Kintzer & Peterson - Primary and Secondary Canine Hypoadrenocorticism
9 pages
Drug Development Process-1
No ratings yet
Drug Development Process-1
12 pages
Gynecology Case Presentation
No ratings yet
Gynecology Case Presentation
10 pages
Nuclear Cardiology
No ratings yet
Nuclear Cardiology
36 pages
The Effect of Hyoscine Butylbromide On The First Stage of Labour in Term Pregnancies
No ratings yet
The Effect of Hyoscine Butylbromide On The First Stage of Labour in Term Pregnancies
5 pages
Understanding Delusional Disorder
No ratings yet
Understanding Delusional Disorder
11 pages
Laporan Bulanan JKN Desember 2022
No ratings yet
Laporan Bulanan JKN Desember 2022
4 pages
Basic Skills In: Interpreting Laboratory Data
No ratings yet
Basic Skills In: Interpreting Laboratory Data
28 pages
BOOST Pharm Exam 2 Review - BLANK
No ratings yet
BOOST Pharm Exam 2 Review - BLANK
20 pages
Drug Repurposing Patent Insights
No ratings yet
Drug Repurposing Patent Insights
8 pages