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Ch. 18 - Practical Applications of Immunology
Tortora - Microbiology: An Introduction 14th Edition
Tortora14th EditionMicrobiology: An IntroductionISBN: 9780138200398Non è quello che usi tu?Cambia libro di testo
Tutti i libri di testoTortora 14th EditionCh. 18 - Practical Applications of ImmunologyProblema 4
Capitolo 14, Problema 4

Use the following choices to answer questions 4 and 5:
a. Anti-Brucella
b. Brucella
c. Substrate for the enzyme
Which is the third step in a direct ELISA test?

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1
Understand the principle of a direct ELISA test: it involves detecting an antigen immobilized on a surface using a labeled antibody specific to that antigen.
Recall the sequence of steps in a direct ELISA: first, the antigen (in this case, Brucella) is attached to the well surface.
Second, a labeled antibody (anti-Brucella) is added, which binds directly to the antigen.
Third, after washing away unbound antibodies, a substrate for the enzyme linked to the antibody is added to produce a detectable signal.
Therefore, identify the third step as the addition of the substrate for the enzyme to develop the signal.

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Direct ELISA Procedure

Direct ELISA is an immunoassay technique used to detect the presence of an antigen in a sample. It involves immobilizing the antigen on a surface, then adding a labeled antibody that directly binds to the antigen. Understanding the sequence of steps—coating, blocking, antibody binding, and detection—is essential to identify the third step.
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Direct Microscopic Cell Counts

Role of Antibodies in ELISA

Antibodies are proteins that specifically bind to antigens. In direct ELISA, a labeled antibody (such as anti-Brucella if detecting Brucella antigen) binds directly to the target antigen. Recognizing the function and specificity of antibodies helps clarify which reagent is added at each step.
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Antibodies

Enzyme-Substrate Reaction in ELISA Detection

The enzyme linked to the antibody catalyzes a reaction with a substrate to produce a detectable signal, usually a color change. This step follows antibody binding and is crucial for visualizing the presence of the antigen. Knowing when the substrate is added helps determine the order of ELISA steps.
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Enzyme-Substrate Complex
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Use the following choices to answer questions 4 and 5:

a. Anti-Brucella

b. Brucella

c. Substrate for the enzyme

Which item is from the patient in an indirect ELISA test?

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The examples in questions 1 and 2 are:

a. Direct tests

b. Indirect tests

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How are monoclonal antibodies produced?

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Explain the effects of excess antigen and antibody on the precipitation reaction. How is the precipitin ring test different from an immunodiffusion test?

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Use the following choices to answer questions 1 and 2:

a. Hemolysis

b. Hemagglutination

c. Hemagglutination inhibition

d. No hemolysis

e. Precipitin ring forms

Patient’s serum, Chlamydia, guinea pig complement, sheep red blood cells, and anti-sheep red blood cells are mixed in a tube. What happens if the patient has antibodies against Chlamydia?

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Label the components of the direct and indirect FA tests in the following situations. Which test is direct? Which test provides definitive proof of disease?

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