Test Bank for Immunology and Serology in Laboratory Medicine 6th Edition by Turgeon

Test Bank for Immunology and Serology in

Laboratory Medicine 6th Edition by Turgeon

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Chapter 10: Precipitation and Particle Agglutination Methods
Turgeon: Evolve Resources for Immunology & Serology in Laboratory Medicine, 6th
Edition

MULTIPLE CHOICE

1. The definition of the term agglutination is the

a. visible expression of the aggregation of antigens and antibodies.
b. formation of a framework in which antigen particles or molecules alternate with
antibody molecules.
c. term used to describe the aggregation of particulate test antigens.
d. All of the above.
ANS: D
Precipitation and agglutination are the visible expressions of the aggregation of antigens and
antibodies through the formation of a framework in which antigen particles or molecules
alternate with antibody molecules. Agglutination is the term used to describe the aggregation
of particulate test antigens.

DIF: Cognitive Level: II REF: p. 146

2. Precipitation is the term applied to

a. the aggregation of soluble test antigens.
b. the aggregation of soluble test antibodies.
c. Either A or B.
d. the aggregation of particulate test antigens.
ANS: A
Precipitation is the term applied to aggregation of soluble test antigens.

DIF: Cognitive Level: II REF: p. 146

3. An artificial carrier could be

a. latex particles.
b. colloidal charcoal.
c. erythrocytes coated with antigen.
d. Both A and B.
ANS: D
Examples of artificial carriers include latex particles and colloidal charcoal. Cells unrelated to
the antigen, such as erythrocytes coated with antigen in a constant amount, can be used as
biological carriers.

DIF: Cognitive Level: I REF: p. 147 | p. 149

4. The quality of agglutination or precipitation test results depends on

a. length of incubation with the serum containing antibody.
b. amount of antigen conjugated to the carrier.
c. pH.
d. All of the above.
 ANS: D
The quality of test results depends on a variety of factors, including the following:
• Time of incubation with the antibody source (i.e., patient serum)
• Amount and avidity of an antigen conjugated to the carrier
• Conditions of the test environment (e.g., pH and protein concentration)

DIF: Cognitive Level: II REF: p. 147 | p. 148

5. All the following procedures are typically performed by latex agglutination or latex
agglutination inhibition assays except
a. C-reactive protein (CRP).
b. rheumatoid factors (IgG).
c. pregnancy testing.
d. RPR for syphilis.
ANS: D
The following immunologic assays are performed by latex particle agglutination:
• Latex agglutination—CRP, IgG and IgM rheumatoid factors, rubella antibody
• Latex agglutination inhibition—pregnancy testing
In latex agglutination procedures, if an antigen is present in a test specimen (e.g., CRP), the
antigen will bind to the combining sites of the antibody exposed on the surface of the latex
beads, forming visible cross-linked aggregates of latex beads and antigen. In some test
systems (e.g., pregnancy, rubella antibody), latex particles can be coated with antigen. In the
presence of serum antibodies, these particles agglutinate into large, visible bodies.
Flocculation testing can be used in syphilis serologic testing. In the rapid plasma reagin (RPR)
test, the antigen, cardiolipin-lecithin-coated cholesterol with choline chloride, contains
charcoal particles that allow for macroscopically visible flocculation.

DIF: Cognitive Level: II REF: p. 147

6. With agglutination inhibition in pregnancy testing, a negative result is evidenced by

a. agglutination.
b. no agglutination.
ANS: A
The determination of in vitro agglutination inhibition depends on incubation of the patient’s
specimen with anti-hCG, followed by the addition of latex particles coated with human
chorionic gonadotropin (hCG). If hCG is present, it neutralizes the antibody; thus, no
agglutination of latex particles is seen. If no hCG is present, agglutination occurs between the
anti-hCG and hCG-coated latex particles.

DIF: Cognitive Level: II REF: p. 150

7. A false-positive latex agglutination assay for pregnancy can be produced by all the following
except:
a. cross-reactivity with luteinizing hormone (LH).
b. cross-reactivity with follicle-stimulating hormone (FSH).
c. antisperm antibodies.
d. monoclonal antibody directed against the beta subunit of hCG.
ANS: D
 False-Positive Results
If a patient has been given an hCG injection (e.g., Profasi, Pregnyl) to trigger ovulation or
lengthen the luteal phase of the menstrual cycle, trace amounts can remain in the patient’s
system for as long as 10 days after the last injection. This will produce a false-positive result.
Two consecutive quantitative hCG blood assays can circumvent this problem. If the hCG
level increases by the second test, the patient is probably pregnant.
Chorioepithelioma, hydatidiform mole, or excessive ingestion of aspirin may yield
false-positive results. In men, a test identical to the one used for pregnancy may be performed
to detect the presence of a testicular tumor. If monoclonal antibody against the beta subunit is
not used, other hormones with the same alpha unit may cross-react and cause a false-positive
reaction.
False-Negative Results
Testing before reaching detectable levels of hCG will yield false-negative results.

DIF: Cognitive Level: II REF: pp. 148-150

8. The first phase of agglutination represents

a. physical attachment of antibody molecules to antigens on the erythrocytic
membrane.
b. physical attachment of antigens to antibody molecules on the erythrocytic
membrane.
c. a reversible chemical reaction.
d. Both A and C.
ANS: D
The first phase of agglutination, sensitization, represents the physical attachment of antibody
molecules to antigens on the erythrocytic membrane. The combination of antigen and
antibody is a reversible chemical reaction. Altering the physical conditions can result in the
release of antibody from the antigen-binding site.

DIF: Cognitive Level: II REF: p. 151 | p. 152

9. The most efficient type of antibodies in agglutination reactions are

a. IgM.
b. IgG.
c. IgD.
d. IgE.
ANS: A
IgM antibodies are more efficient at agglutination because their large size and multivalency
permit more effective bridging of the space between cells caused by zeta potential. IgG
antibodies are too small to overcome electrostatic forces between cells.

DIF: Cognitive Level: II REF: p. 152

10. Anti-human globulin (AHG) is used to

a. substitute for a low titer of antibody.
b. alter the configuration of an antigen.
c. form cross-links between antibodies bound to the surface of erythrocytes.
d. form cross-links between antigens bound to the surface of erythrocytes.
 ANS: C
The use of AHG forms cross-links between antibody molecules that have bound to the surface
of red blood cells. This promotes the formation of agglutination and allows visual observation
of an antigen-antibody reaction.

DIF: Cognitive Level: II REF: p. 153

11. The lattice hypothesis states that an optimal reaction occurs between antigen and antibody
when
a. an excess of antigen is present.
b. an excess of antibody is present.
c. antigen and antibody are present in approximately equal proportions.
d. reaction takes place at room temperature.
ANS: C
Precipitation reactions depend on a zone of equivalence, the zone in which optimum
precipitation occurs because the number of multivalent sites of antigen and antibody are
approximately equal. For a precipitation reaction to be detectable, the reaction must occur in
the zone of equivalence. In this zone, each antibody or antigen binds to more than one antigen
or antibody, respectively, forming a stable lattice or network.
The lattice hypothesis is based on an assumption that each antibody molecule must have at
least two binding sites, and an antigen must be multivalent.

DIF: Cognitive Level: II REF: p. 153

12. The strength of an agglutination reaction can be enhanced by

a. centrifugation.
b. use of albumin.
c. use of low ionic strength saline.
d. All of the above.
ANS: D
Techniques used to enhance agglutination include centrifugation, treatment with proteolytic
enzymes, use of colloids, and AHG testing.

DIF: Cognitive Level: II REF: p. 152

13. If a slide agglutination test is rotated for less than 1 minute, as the directions specify, what
source of error can occur?
a. Possible false-positive
b. Possible false-negative
c. Depends on the amount of antibody present in the sample
d. No effect on the results
ANS: B
Causes of False-Negative Agglutination Reactions
Inadequate washing of red blood cells in antihuman globulin (AHG) testing may result in
unbound immunoglobulins neutralizing the reagent.
Failure to add AHG reagent.
Contaminated or expired reagents.
Improper incubation.
 Delay in reading slide reactions.
Under centrifugation.
Prozone phenomenon.

DIF: Cognitive Level: II REF: p. 150

14. In a group A patient’s reverse serum testing, the cells and serum agglutinate because
a. group A antigen is present on the patient’s erythrocytes.
b. group A antigen is present in the patient’s serum.
c. anti-group A is present in the patient’s serum.
d. Both A and C.
ANS: A
Reactions of Patient Serum and Reagent Erythrocytes

DIF: Cognitive Level: III REF: p. 157

15. In nephelometric methods, the relationship between the protein being assayed (the antigen in
this instance) and the concentration of the protein in the sample (as measured by the
instrument as the amount of scattered light) is
a. inversely proportional.
b. directly proportional.
ANS: B
The procedure is based on the reaction between the protein being assayed and a specific
antiserum. The resulting complexes of insoluble precipitants scatter incident light in solutions.
The amount of scattered light is proportional to the number of insoluble complexes and can be
quantitated by comparing the unknown patient values with standards of known protein
concentration.

DIF: Cognitive Level: II REF: p. 154

16. Latex particles act as ______________________ in nephelometry.

a. antigen-coated reaction intensifiers
b. antigen-coated reaction stabilizers of precipitate
c. sensitivity enhancers
d. color reaction intensifiers
ANS: A
Specific antigen-coated latex particles acting as reaction intensifiers are agglutinated by their
corresponding antibody.

DIF: Cognitive Level: II REF: p. 154

17. A fundamental prerequisite for nephelometric protein quantitation is the
a. dissolution of large protein molecules.
b. formation of a macromolecular complex.
c. pH of the solution.
d. ionic strength of the solution.
ANS: B
Increased light scatter of a solution can be measured by nephelometry as the macromolecular
complexes form.

DIF: Cognitive Level: II REF: p. 154

18. In nephelometry measurement, if antigen overwhelms the quantity of antibody, the measured
signal
a. increases.
b. decreases.
c. remains the same.
d. fluctuates between high and low.
ANS: B
The relationship between the quantity of antigen and measuring signal at a constant antibody
concentration is expressed by the Heidelberger curve. If antibodies are present to excess, a
proportional relationship exists between the antigen and resulting signal. If the antigen
overwhelms the quantity of antibody, the measured signal decreases.

DIF: Cognitive Level: II REF: p. 154

19. Which of the following is a correct statement regarding measurements of turbidity?

a. Light is scattered at a determined angle.
b. It is unaffected by large particles precipitating out of a solution.
c. The reference value is recorded in units of relative light scatter.
d. It represents the scattered light in forward direction.
ANS: A
Nephelometry is based on the principle that light is scattered by a homogeneous particulate
solution at a variety of angles. Three types of scatter can occur: (1) scatter around the
particles; (2) forward scatter caused by out of phase backscatter; and (3) forward scatter
exceeding backscatter.

DIF: Cognitive Level: II REF: p. 154

20. In nephelometry measurement, a __________________ method is used.

a. variable-time
b. fixed-time
c. fluorescent-tagging
d. competitive immunoassay
ANS: B
A fixed-time method is used routinely for precipitation reactions.

DIF: Cognitive Level: II REF: p. 156

 21. Nephelometry can be used to assay all the following except:
a. IgM.
b. IgG.
c. IgD.
d. IgA.
ANS: C
Currently, instruments using a rate method and fixed-time approach are commercially
available with tests for immunoglobulin G (IgG), IgA, IgM, C3, C4, properdin, C-reactive
protein (CRP), rheumatoid factor, ceruloplasmin, 1-antitrypsin, apolipoproteins, and
haptoglobins.

DIF: Cognitive Level: II REF: p. 156

22. Cryoglobulins are proteins that precipitate or gel when cooled to

a. –18° C.
b. 0° C.
c. 4° C.
d. 18° C.
ANS: B
Cryoglobulins are proteins that precipitate or gel when cooled to 0° C and dissolve when
heated.

DIF: Cognitive Level: II REF: p. 157

23. Monoclonal cryoglobulins are most often

a. IgM.
b. IgG.
c. IgA.
d. Either A or B.
ANS: D
In most cases, monoclonal cryoglobulins are IgM or IgG.

DIF: Cognitive Level: II REF: p. 157

24. Cryoglobulin analysis can be useful in the diagnosis of

a. hypothermia.
b. Raynaud’s phenomenon.
c. infectious hepatitis.
d. rheumatoid arthritis.
ANS: B
Cryoglobulin analysis is frequently requested when patient symptoms such as pain, cyanosis,
Raynaud’s phenomenon, and skin ulceration on exposure to cold temperatures are present.

DIF: Cognitive Level: II REF: p. 157

MATCHING

Match the following terms with the correct definition.

 a. Uses antibodies bound to a particle to enhance the visibility of the reaction
b. Based on the interaction of soluble antigen with antibody
c. Detects antibodies with erythrocyte antigens

1. Coagglutination
2. Flocculation
3. Hemagglutination

1. ANS: A DIF: Cognitive Level: II REF: p. 147

2. ANS: B DIF: Cognitive Level: II REF: p. 150
3. ANS: C DIF: Cognitive Level: II REF: p. 151

Match the term with the appropriate description.

a. The zone in which optimum precipitation occurs
b. An excessive antibody concentration that produces a false-negative reaction
c. An excessive antigen concentration that results in no lattice formation
d. An equal amount of antigen and antibody concentrations

4. Prozone phenomenon
5. Postzone phenomenon

4. ANS: B DIF: Cognitive Level: II REF: p. 152 | p. 153

5. ANS: C DIF: Cognitive Level: II REF: p. 152 | p. 153

Match the type of immunoglobulin with the optimal thermal range.

a. 4 to 22 C
b. 18 to 22 C
c. 37 C
d. 56 C

6. IgM
7. IgG

6. ANS: A DIF: Cognitive Level: II REF: p. 153

7. ANS: C DIF: Cognitive Level: II REF: p. 153

Match the cause of a false-positive agglutination reaction with the appropriate corrective
measure.
 a. Use a control with saline and no antibody.
b. Read reactions exactly as specified.
c. Calibrate centrifuge to proper speed and time.
d. Use positive and negative QC cells.
8. Over centrifugation of cells and serum
9. Autoagglutination

8. ANS: C DIF: Cognitive Level: III REF: p. 150 | p. 153

9. ANS: A DIF: Cognitive Level: III REF: p. 150 | p. 153

Match the cause of a false-negative agglutination reaction with the appropriate corrective
measure.

a. Dilute patient’s serum and retest.

b. Wash cells according to directions; use positive and negative quality control (QC)
steps.
c. Repeat blood specimen should be collected a week or more later.
d. Calibrate centrifuge to proper speed and time.
Test Bank for Immunology and Serology in Laboratory Medicine 6th Edition by Turgeon

10. Prozone phenomenon

11. Anti-human globulin (AHG) malfunction
12. Postzone phenomenon

10. ANS: A DIF: Cognitive Level: III REF: p. 152 | p. 153

11. ANS: B DIF: Cognitive Level: III REF: p. 152 | p. 153
12. ANS: C DIF: Cognitive Level: III REF: p. 152 | p. 153

Match the grade of the agglutination reaction with the appropriate description.

a. Tiny aggregates
b. No aggregates
c. Medium-sized aggregates, some free red blood cells (RBCs)
d. Clear supernatant
13. Negative
14. Weak ()
15. 2+
16. 4+

13. ANS: B DIF: Cognitive Level: I REF: p. 153 | p. 154

14. ANS: A DIF: Cognitive Level: I REF: p. 153 | p. 154
15. ANS: C DIF: Cognitive Level: I REF: p. 153 | p. 154
16. ANS: D DIF: Cognitive Level: I REF: p. 153 | p. 154

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