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HDM Final > Immunological Diagnostics > Flashcards

Immunological Diagnostics Flashcards

1
Q

When are serological assays used for immuological diagnostics?

A

When pathogen is difficult, dangerous, or impossible to culture

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2
Q

What must be available for serological assays for immunological diagnostics?

A

The antibody or antigen in pure, detectable form

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3
Q

What are the 4 most important characteristics of antibody binding used in serological assays?

A

Specificity
Amount
Isotype
Affinity

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4
Q

____________ is an important property of an immunodiagnostic test referring to the ability to detect small quantities of Ab or Ag

A

Sensitivity

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5
Q

An immunodiagnostic test with poor sensitivity would result in which of the following?

A. False positives
B. True positives
C. False negatives
D. True negatives

A

C. False negatives

[lack of detection when Ab or Ag is present]

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6
Q

What can you conclude about disease status if the sensitivity of an immunodiagnostic assay is HIGH?

A

You can rule OUT disease

[if high sensitivity test is positive, may want to confirm with a highly specific test, because it is possible you got a false positive]

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7
Q

__________ is an important property of an immunodiagnostic test referring to the ability to distinguish the immunogen from all other similar antigens

A

Specificity

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8
Q

An immunodiagnostic test with poor specificity would result in which of the following?

A. False positives
B. True positives
C. False negatives
D. True negatives

A

A. False positives

[detects something other than the immunogen of interest]

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9
Q

What can you conclude about disease status if the specificity of an immunodiagnostic assay is HIGH?

A

You can rule IN disease

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10
Q

What type of serological assay results in the formation of a complex that visibly precipitates?

A

Precipitin reactions

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11
Q

What type of serological assay results in cells/particles bound together through antibody-antigen interactions?

A

Agglutination reactions

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12
Q

What type of serological assay results in antibody covalently-linked to a label?

A

Labeled antibody methods

[antibody bound to Ag is detected by presence of the label]

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13
Q

What is the difference between direct and indirect serological assay methods?

A

Direct detects ANTIGEN

Indirect detects ANTIBODY

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14
Q

What are some other terms used for antibody?

A

Antiserum
Serum
Antitoxin
Antivenin

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15
Q

What is the direct fluorescent Ab test?

A

Identifies Ag in tissue by labeling monoclonal Abs specific to the epitope you are looking for

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16
Q

In the direct fluorescent Ab test, where on the Ab does the label stick?

A

The Fc region of IgG

[if this Ab binds Ag of interest, you will see fluorescence on microscopy]

17
Q

What is the indirect fluorescent Ab test?

A

Identifies Ab in patient

[make test Ag that patient’s fluorescent-labeled Abs will bind to]

18
Q

In precipitin reactions, precipitation occurs when _________ antigens and antibodies ______-______, forming large molecular aggregates

A

Multivalent; cross-link

19
Q

For precipitin reactions, what is the term for the point at which precipitation occurs?

A

Zone of equivalence (or equivalence point)

20
Q

Prior to reaching the zone of equivalence in a precipitin reaction, what is in excess - antibody or antigen?

A

Antibody

21
Q

Once you are past the zone of equivalence in a precipitin reaction, what is in excess - antibody or antigen?

A

Antigen

22
Q

What serological test is used to determine if two antigens contain overlapping epitopes?

What is this test also used for?

A

Ouchterlony double diffusion

[also used to determine concentrations of antigens]

23
Q

Agglutination reactions are similar to precipitin reactions, but what is the difference in agglutination reactions?

A

In agglutination reactions, the antigen is part of a whole cell or cell-sized particle

24
Q

Describe the degree of sensitivity and speed in an agglutination reaction

A

Very sensitive

Very fast reaction - occurs in minutes

25
Q

_____________ is the agglutination of erythrocytes and is used for blood typing

A

Hemagglutination

26
Q

What test uses anti-human immunoglobulins to precipitate agglutination of RBCs bound to anti-Rh antibodies?

A

The coombs test

27
Q

What does the direct Coombs test detect?

A

Detects anti-Rh antibodies found on fetal RBCs

28
Q

What does the indirect coombs test detect?

A

Detects anti-Rh antibodies in mother’s serum

29
Q

Which of the following best describes the Coombs test?

A. Precipitin reaction
B. Agglutination reaction
C. Labeled antibody method
D. Labeled antigen method

A

B. Agglutination reaction

30
Q

Labeled Ab methods detect antibodies bound to a _________ surface.

Antibodies are labeled in _______ amounts to ensure binding to Ag is not interrupted. The label is added to the ____ region of the Ab.

Direct and indirect labeling of antibodies is done, but _______ labeling is more popular.

A

Solid

Small; Fc

Indirect

31
Q

What test involves labeled antibodies (linked to enzyme) added to wells coated wtih different antigens?

A

ELISA

[antibodies specific to Ag will bind, while those that do not recognize Ag will be washed away]

32
Q

How is Ag:Ab binding detected in ELISA tests?

A

Measuring color change, mediated by enzyme-dependent conversion of a colorless substrate to a colored reactant

33
Q

What type of blotting technique is sometimes used following a positive ELISA to confirm the findings?

A

Western blot (looks for protein)

34
Q

Western blot is used to confirm diagnosis of HIV. The HIV particle is dissociated by ______ and separated on a _______.

Anti-HIV Abs in the serum of an infected individual will bind to the various HIV proteins. Labeled anti-human ______ are used to detect antibodies bound to the HIV proteins

A

SDS; PAGE

Immunoglobulins

35
Q

What technique is used as a tool for defining and enumerating lymphocytes using antibodies specific for cell-surface proteins?

A

Flow cytometry

[the flow cytometer detects and counts individual cells passing in a stream through a laser beam]

36
Q

Flow cytometers are equipped to identify what aspect of cells?

A

Their cell-surface antigens (it sorts them accordingly)

37
Q

Which type of T lymphocyte is more numerous, CD4 or CD8?

A

CD4

38
Q

What technique is used for detection of lymphocyte subpopulations using specific antibodies?

A

Fluorescence-activated cell sorting (FACs) - a specialized type of flow cytometry that sorts a heterogenous mixture of cells, one cell at a time, based on specific light scattering and fluorescent characteristics of each cell

HDM Final (12 decks)

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