Unzipping Genes Product Information

HiPer® Blood Grouping Teaching Kit

Product Code: HTI008

Number of experiments that can be performed: 100

Duration of Experiment:
Protocol: 15 minutes

Storage Instructions:
 The kit is stable for 12 months from the date of manufacture
 Store Anti A, Anti B and Anti RhD at 2-8oC
 Other kit contents can be stored at room temperature (15-25oC)

Registered Office : Commercial Office

WHO 23, Vadhani Industrial Estate,LBS Marg, A-516, Swastik Disha Business Park, Tel: 00-91-22-6147 1919
15 Mumbai - 400 086, India. Fax: 6147 1920, 2500 5764
GMP Tel. : (022) 4017 9797 / 2500 1607
Via Vadhani Indl. Est., LBS Marg,
Mumbai - 400 086, India Email : info@himedialabs.com
CERTIFIED Fax : (022) 2500 2286 Web : www.himedialabs.com

The information contained herein is believed to be accurate and complete. However no warranty or guarantee whatsoever is made
or is to be implied with respect to such information or with respect to any product, method or apparatus referred to herein
 Index

Sr. No. Contents Page No.

1 Aim
3
2 Introduction
3
3 Principle
3
4 Kit Contents
4
5 Materials Required But Not Provided
4
6 Storage
4
7 Important Instructions
4
8 Procedure
4
9 Observation and Result
5
1 0 Interpretation
6
1
1 Troubleshooting Guide
6

2
Aim:

To determine the blood group and Rh factor of an individual.

Introduction:

Blood grouping is the classification of blood based on the presence or absence of two inherited antigenic
substances on the surface of red blood cells (RBCs). The ABO and Rh are the major, clinically significant and
the most important of all the blood group systems. The ABO blood group system was first discovered by Karl
Landsteiner in 1 900. The human ABO blood group system is divided into the following four major groups
depending on the antigen present on the surface of their red blood cells:
1. “A” group
2. “B” group
3. “AB” group
4. “O” group

Table 1: ABO Blood Group System

Antigens on the surface of Red Antibodies in the Serum ABO Blood Group Genotype
Blood Cells

A Anti B A AA or AO
B Anti A B BB or BO
A and B Neither Anti A nor Anti B AB AB
Neither A nor B Anti A, Anti B, Anti AB O OO

The associated Anti A and Anti B antibodies usually belong to IgM class of immunoglobulins.
The Rhesus system (Rh) is the second most important blood group system in humans. The most significant
and immunogenic Rhesus antigen is the RhD antigen. The individuals carrying the Rh antigen are considered
to have positive blood group whereas those individuals that lack this antigen are considered to have negative
blood group.

Principle:

The ABO and Rh blood grouping system is based on agglutination reaction. When red blood cells carrying one
or both the antigens are exposed to the corresponding antibodies they interact with each other to form visible
agglutination or clumping. The ABO blood group antigens are O-linked glycoproteins in which the terminal
sugar residues exposed at the cell surface of the red blood cells determine whether the antigen is A or B. Blood
group A individuals have A antigens on RBCs and anti-B antibodies in serum. Similarly, blood group B
individuals have B antigens on RBCs and anti-A antibodies in serum. Blood group AB individuals have both A
and B antigens on RBCs and neither anti-A nor anti-B antibodies in serum. Whereas, blood group O individuals
have neither A antigens nor B antigens, but possess both anti-A and anti-B antibodies in serum. The Rh antigens
are transmembrane proteins in which the loops exposed on the surface of red blood cells interact with the
corresponding antibodies.

3
Kit Contents:

HiPer® Blood Grouping Teaching Kit enables rapid identification of ABO blood group and Rh factor depending
upon the antigen present on the surface of red blood cells.

Table 2: Enlists the materials provided in this kit with their quantity and recommended storage

Product Quantity
Sr. No. Materials Provided Storage
Code 100 expts
1 TKC104 Anti A Sera 5 ml 2-8oC
2 TKC105 Anti B Sera 5 ml 2-8oC
3 TKC106 Anti RhD Sera 5 ml 2-8oC
4 TKC109 Cavity slide 10 Nos. RT
5 TKC125 Disposable Mixing Stick 300 Nos. RT
6 1
TKC 08 Blood Lancet 100 Nos. RT

Materials Required But Not Provided:

Reagents: 70% Alcohol/ Spirit

Other requirements: Cotton

Storage:

HiPer® Blood Grouping Teaching Kit is stable for 12 months from the date of manufacture without showing any
reduction in performance. Store the Anti A Sera, Anti B Sera and Anti RhD Sera at 2-8oC. Other contents can be
stored at room temperature (15-25oC).

Important Instructions:

1. Before starting the experiment the entire procedure has to be read carefully.
2. Always wear gloves while performing the experiment.
3. Ensure the slide is clean and dry prior to use.
4. Do not allow the antisera reagent dropper to touch the blood sample.
5. The result of the reaction should be interpreted immediately after mixing.
6. Avoid intermixing of the antisera reagents while performing the experiment as it may
give false result.

Procedure:

1. Dangle the hand down to increase the flow of blood in the fingers.
2. Clean the fingertip to be pierced with spirit or 70% alcohol (usually ring or middle finger).
3. With the help of the sterile lancet, pierce the fingertip and place one drop of blood in each of the cavities.

4
4. Add one drop of antiserum into each cavity as shown below:

Blood drop Blood drop Blood drop

+ + +
Anti A Anti B Anti RhD

5. Mix each blood drop and the antiserum using a fresh mixing stick.
6. Observe agglutination in the form of fine red granules within 30 seconds. Anti RhD takes slightly longer
time to agglutinate compared to Anti A and Anti B.

Note: Proper care should be taken while disposing the lancet and mixing sticks.

Observation and Result:

A B D A B D

Slide 1 Slide 2

A B D A B D

Slide 3 Slide 4

Table 3: Determination of blood group and Rh factor based on agglutination seen

Sr. No. Anti A Anti B Anti RhD Blood Group

Slide 1    A +ve
Slide 2    B +ve
Slide 3    AB +ve
Slide 4    O +ve

: Agglutination
: No agglutination

5
 Interpretation:

 If agglutination is observed when blood is mixed with Anti A reagent, then the individual is said to
have blood group “A”.
 If agglutination is observed when blood is mixed with Anti B reagent, then the individual is said to
have blood group “B”.
 If agglutination is observed when blood is mixed with Anti A and Anti B reagent, then the individual
is said to have blood group “AB”.
 If no agglutination is observed when blood is mixed with Anti A and Anti B reagent, then the
individual is said to have blood group “O”.
 If agglutination is observed when blood is mixed with Anti RhD reagent, then the individual is said to
have “+ve” Rh factor.
 If no agglutination is observed when blood is mixed with Anti RhD reagent, then the individual is
said to have “-ve” Rh factor.

Troubleshooting Guide:

Sr.No Problem Possible Cause Solution

Ensure that the antisera reagents are added

The antisera reagents
properly onto the respective cavity without spilling
mix with each other
1. False positive to the sides
result
Incubated for a longer The results should be read within the time period
time mentioned in the brochure

Antisera not stored Ensure that the antisera is stored in refrigerator

No agglutination
2. under proper (2-8oC)
observed
conditions

6
Technical Assistance:

At HiMedia we pride ourselves on the quality and availability of our technical support. For any kind of
technical assistance, mail at mb@himedialabs.com

25°C Storage temperature

15°C

Do not use if package is

damaged

HiMedia Laboratories Pvt. Limited,

23 Vadhani Industrial Estate,
LBS Marg,Mumbai-86,MS,India

PIHTI008_O/0419 HTI008-04