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Chapter 7: Antibody Detection and Identification
Test Bank

MULTIPLE CHOICE

1. In the process of identifying an antibody, the technologist observed 2+ reactions with 3 of the
10 cells in a panel at the immediate spin phase. These reactions disappeared following
incubation at 37 C and the antihuman globulin phase of testing. The antibody most likely to
be responsible is:
a. anti-E.
b. anti-D.
c. anti-I.
d. anti-Lea.
ANS: D
The Lea antigen is typically found on three to four cells in a panel, and the antibody reacts best
at the immediate phase of testing.

DIF: Level 3 REF: p. 173

2. Rh system antibodies characteristically give:

a. mixed-field reactions on panels.
b. weak reactions with panel cells.
c. strong reactions with panel cells when read at immediate spin phase.
d. reactions that are enhanced with enzymes.
ANS: D
Rh system antibodies are typically strong and are enhanced with enzyme treatment.

DIF: Level 1 REF: p. 168

3. Which of the following situations can be found in a classic case of autoimmune hemolytic
anemia?
a. Positive direct antiglobulin test
b. False-positive Fya phenotyping
c. Crossmatch incompatibility at antihuman globulin
d. All of the above
ANS: D
Red cells of a patient with this condition are coated with IgG antibodies that are signaling
premature destruction in the spleen. Because the red cells are coated, attempts to test them
with antiglobulin reagent results in positive reactions, and the antibody in the serum will react
with all normal cells tested at the antihuman globulin phase.

DIF: Level 2 REF: p. 177

4. The next step in investigating a positive direct antiglobulin test using polyspecific antihuman
globulin reagent should be to:
a. repeat the direct antiglobulin test using warm saline.
b. perform an eluate.
 c. add IgG-sensitized red cells to verify positive reaction.
d. repeat the direct antiglobulin test using monospecific anti-IgG and anti-C3
reagents.
ANS: D
Polyspecific antihuman globulin contains both a complement and IgG component. To
determine which caused the positive reaction, red cells should be tested separately using
monospecific anti-IgG and anti-C3 reagents.

DIF: Level 2 REF: p. 160

5. Antibody screening cells are positive at the antihuman globulin phase of testing. The first step
of the investigation should be to:
a. check transfusion and pregnancy history.
b. perform a direct antiglobulin test using anti-C3.
c. repeat the ABO typing.
d. crossmatch units until one is compatible.
ANS: A
A positive screen indicates the presence of unexpected antibody. Patient history can aid in the
investigation.

DIF: Level 2 REF: p. 161

6. The phase of the agglutination reaction is important in the interpretation of the antibody
screen or antibody identification panel because it:
a. determines whether there is a delayed transfusion reaction.
b. provides clues on antibody dosage.
c. indicates the class of the antibody.
d. determines whether an autoantibody is present.
ANS: C
IgM antibodies typically react at room temperature. IgG antibodies require the antiglobulin
phase to detect.

DIF: Level 2 REF: p. 165

7. In an antibody identification panel, only one red cell was negative at the antihuman globulin
phase. On ruling out and matching the pattern, an anti-k was identified. What further testing is
necessary to confirm the antibody?
a. Two more k-negative cells should be tested.
b. Two more K-negative cells should be tested.
c. Treat the panel cells with enzymes and perform the panel again.
d. Perform an adsorption using “k”-positive cells.
ANS: A
To satisfy the “rule of three,” three negative and three positive reactions for the antigen should
be observed to rule in an antibody.

DIF: Level 3 REF: p. 166

 8. An anti-Fya was identified in a patient’s serum. The patient’s red cells phenotyped as Fya
positive using commercial antisera. The next step is to:
a. repeat the panel to confirm the antibody.
b. report the antibody because this result is normal.
c. investigate a recent transfusion history.
d. wash the cells and use monoclonal anti-Fya antibodies.
ANS: C
The patient’s red cells should be Fya negative to make anti-Fya unless a recent transfusion was
given of Fya positive red cells. If no Fya-positive red cells were recently transfused, the
antibody identified may be incorrect or the patient may have a positive direct antiglobulin test.

DIF: Level 3 REF: p. 166

9. If all the panel cells were reactive at the same strength at the antihuman globulin phase, no
negative reactions were observed, and the autocontrol was negative, what should be
suspected?
a. Multiple antibody specificities
b. Warm autoantibody
c. Antibody to a low-frequency antigen
d. Antibody to a high-frequency antigen
ANS: D
Reactions of similar strength suggest one specificity; a negative autocontrol rules out an
autoantibody, and all panel cells reactive suggest an antibody to an antigen that is of high
frequency in the population.

DIF: Level 3 REF: p. 169

10. Antibodies to low-incidence antigens include all of the following except:

a. anti-Vel.
b. anti-Cw.
c. anti-V.
d. anti-Lua.
ANS: A
Vel is an antigen found in high frequency in the population.

DIF: Level 1 REF: p. 171

11. Cold autoantibodies are typically of which specificity?

a. M
b. N
c. I
d. Leb
ANS: C
Cold autoantibodies are typically of the specificity anti-I. I is a high-frequency antigen found
on all adult red cells but is absent on cord red cells.

DIF: Level 1 REF: p. 174

12. If an anti-I is suspected in a patient’s sample that requires a transfusion, the most acceptable
course of action is to:
a. call the rare donor registry.
b. crossmatch cord blood.
c. perform a cold autoadsorption.
d. perform the prewarm technique.
ANS: D
Prewarming the patient’s serum and panel cells separately and then mixing at 37° C to avoid
cold temperatures usually avoids the reactivity of the anti-I, which is not clinically significant
but may mask other antibodies.

DIF: Level 2 REF: p. 176

13. What is the most important concern when trying to identify antibodies in a patient with a
warm autoantibody?
a. Identifying the specificity of the autoantibody
b. Determining whether there are underlying alloantibodies
c. Identifying the antibody found in the eluate
d. Determining whether complement is binding to the autologous red cells
ANS: B
Autoantibodies in the serum can mask underlying alloantibodies.

DIF: Level 2 REF: p. 178

14. An autoadsorption may be performed to investigate underlying autoantibodies. When is this

procedure acceptable?
a. When the autoantibody is reactive at 4° C
b. When the patient has not been recently transfused
c. Only if complement is coating the red cells
d. When the eluate is negative
ANS: B
If an autoadsorption is performed (using the patient’s red cells) and the patient has been
recently transfused, the adsorbing red cells may remove developing alloantibodies to the
transfused cells.

DIF: Level 2 REF: p. 180

15. Proteolytic enzymes should not be used to screen for antibodies because:
a. the reagent is too expensive for routine use.
b. clinically insignificant antibodies are enhanced.
c. red cells must be treated with enzymes first, which makes this technique
impractical.
d. some antigens are destroyed by enzymes, which would cause the antibodies to be
missed.
ANS: D
Proteolytic enzymes destroy some antigens in the Duffy and MNS system. Antibody screens
using enzymes would not detect antibodies to these antigens.
 DIF: Level 1 REF: p. 161

16. High-titer, low-avidity antibodies typically:

a. react with antigens of high frequency in the population.
b. react with antigens of low frequency in the population.
c. are clinically significant.
d. react best at colder temperatures.
ANS: A
High-titer, low-avidity antibodies are typically reactive with most panel cells at the antihuman
globulin phase and are not clinically significant. They may mask clinically significant
reactions.

DIF: Level 1 REF: p. 170

17. An example of a cold alloantibody includes:

a. anti-M.
b. anti-I.
c. anti-Lub.
d. anti-k.
ANS: A
Alloantibodies to anti-M react best at room temperature but can also demonstrate reactions at
37° C and antihuman globulin phases.

DIF: Level 1 REF: p. 172

18. The best description of the elution technique is that it is a technique used to:
a. disassociate IgM antibodies from red cells for further identification.
b. disassociate IgG antibodies from red cells for further identification.
c. adsorb IgG antibodies from serum.
d. separate IgG and IgM antibodies in serum.
ANS: B
Elution procedures remove IgG antibodies from sensitized red cells to be used for
identification using panel cells.

DIF: Level 2 REF: p. 179

19. All the following antigens are commonly found on screening cells except:
a. D.
b. k.
c. Kpa.
d. C.
ANS: C
Kpa is a low-frequency antigen in the Kell system that is typically not present on screening
cells.

DIF: Level 1 REF: p. 159

20. An autoadsorption uses what type of cells to remove antibody from the serum?
 a. Antibody screening cells
b. Donor red cells
c. Patient red cells
d. Antibody identification panel cells
ANS: C
Patient red cells are treated to remove IgG antibody and then are incubated with the patient’s
serum to remove more autoantibodies that are interfering with alloantibody identification.

DIF: Level 1 REF: p. 177

21. Anti-D, anti-K, and anti-Jka are the antibodies that are tentatively identified on a panel after
initially ruling out on negative cells. What selected cell from another panel should be chosen
to confirm the presence of anti-K?
a. K–, D+, Jk(a+)
b. K+, D+, Jk(a+)
c. K+, D–, Jk(a+)
d. K+, D–, Jk(a–)
ANS: D
To confirm the presence of the anti-K, a cell positive for K antigen and antigen negative for
the other two suspected antibodies will confirm anti-K if it is reactive against it.

DIF: Level 2 REF: p. 168

22. DTT (dithiothreitol) would be useful in the identification of which of the following
antibodies?
a. Anti-Jsa
b. Anti-Kpb
c. Anti-Vel
d. Anti-K
ANS: B
Anti-Kpb is a high-frequency antigen in the Kell system. If an Anti-Kpb is suspected, the panel
cells could be treated with DTT and the sample retested. If all reactions are eliminated, the
anti-Kpb is confirmed and other underlying alloantibodies are ruled out. Although K would
also be destroyed by DTT, there are sufficient negative cells on the panel to determine if
underlying antibodies exist and confirm the specificity.

DIF: Level 1 REF: p. 170

23. Anti-Sda should be suspected if:

a. weak reactions at the antiglobulin phase occur with several panel cells.
b. reactions are stronger with enzymes at the immediate spin phase.
c. the antibody reacts with most panel cells and are mixed field and refractile
microscopically.
d. weak reactions at the AHG phase titer out to high dilutions.
ANS: C
Anti-Sda antibodies are characteristically mixed field and refractile when observed under the
microscope. Sda is a high-frequency antigen; therefore, these reactions will be observed with
most cells tested.
 DIF: Level 2 REF: p. 170

24. A patient’s serum reacted weakly with all panel cells tested at the antiglobulin phase using
LISS and were not enhanced using PEG. The autocontrol was negative. Ficin-treated panel
cells were nonreactive. What is the most likely specificity of the antibody?
a. Anti-I
b. Anti-U
c. Anti-Ch
d. Anti-Jsb
ANS: C
Anti-Ch (anti-Chido) is an antibody to a high-frequency antigen in the high-titer low-avidity
category of antibodies that demonstrate weak reactions that are not normally enhanced with
potentiators. Anti-Ch reactions may be eliminated when testing with ficin-treated cells.

DIF: Level 2 REF: p. 170

25. Which of the following medications is most likely to cause the production of autoantibodies?
a. Tetracycline
b. Cephalothin
c. Methyldopa
d. Acutane
ANS: C
Methyldopa is medication often associated with autoantibody formation and can be observed
in the serum and the eluate without the presence of the drug (drug-independent mechanism).

DIF: Level 2 REF: p. 179

26. An antibody was detected at immediate spin and 37° C that appeared to have anti-Leb
specificity. To confirm the antibody identity and determine if there were other antibodies in
the serum, a Lewis neutralization technique was performed. Results of the control are as
follows:
Reaction with Le(b) positive cells
Patient serum + Lewis substance 0
Patient serum + saline control 1+
What conclusion can be made from these results?
a. anti-Leb is confirmed
b. the antibody was diluted, therefore no conclusion can be made
c. the antibody was not neutralized, therefore anti-Leb has not been identified
d. an antibody other than anti-Leb is most likely in the serum
ANS: A
Since the saline control demonstrated a positive reaction, the antibody was not diluted by the
neutralization procedure. The antibody to the Lewis antigen was neutralized since it did not
react with the Lewis positive cells.

DIF: Level 3 REF: p. 173

MATCHING
 Match the tentative interpretation of antibody screen and direct antiglobulin test (DAT) with
the results given below.
a. Alloantibody, IgG
b. Alloantibody, IgM
c. Autoantibody, IgM
d. Autoantibody or transfusion reaction, IgG

1. All screening cells 2+ at antihuman globulin phase, DAT positive, IgG 2+

2. One screening cell 1+ at antihuman globulin phase, DAT negative
3. All screening cells positive 1+ at IS, DAT positive, C3 1+
4. All screening cells positive 1+ at IS, DAT negative

1. ANS: D DIF: Level 2

2. ANS: A DIF: Level 2
3. ANS: C DIF: Level 2
4. ANS: B DIF: Level 2
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