Laboratory Experiments in Microbiology (11th Edition)
11th Edition
ISBN: 9780321994936
Author: Ted R. Johnson, Christine L. Case
Publisher: PEARSON
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Textbook Question
Chapter 44, Problem 1Q
Was the test performed in this exercise a direct or an indirect ELISA test?
Expert Solution & Answer
Summary Introduction
To fill:
Indirect ELISA.
Introduction:
Immunoassays are tests done for the diagnosis of diseases by identifying the presence of antigen or a particular antibody from a patient's sample. Antigen-Antibody interactions form the basis of immunoassays wherein interactions like agglutinations, precipitation is seen. Immunoassays using enzyme labels are termed as ELISA-Enzyme Linked ImmunoSorbent Assay.
Answer to Problem 1Q
The test performed in this exercise is indirect ELISA.
Explanation of Solution
In Indirect ELISA first the antigen is added to the well and then the primary antibody is added. After the buffer wash, enzyme-labeled secondary antibody is added which binds to the Antigen-primary antibody complex. Finally, the substrate corresponding to the enzyme is added, which forms a color emitting enzyme-substrate complex. The color change indicates that appropriate Antibody is present in the patient's sample.
In this exercise, initially, S.entericaTyphimurium (antigen) is added, and the patient's serum (i.e primary antibody) is added followed by the addition of alkaline phosphatase labeled with secondary antibody. Finally, the alkaline phosphatase substrate (BCIP/NBT) is added in order to give color if there is an interaction between the antigen and the primary antibody.
Hence the test followed in the exercise is Indirect ELISA.
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Chapter 44 Solutions
Laboratory Experiments in Microbiology (11th Edition)
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