Serological Testing Lab Report: Indirect ELISA Insights

PhysioEx Lab Report Exercise 12: Serological Testing Activity 3: Indirect Enzyme-Linked Immunosorbent Assay (ELISA) Name: Jennifer Adams Date: 4 May 2022 Session ID: session-afab5a2b-cc34-0bd1-4b32-9cdaf0486971 Pre-lab Quiz Results You scored 100% by answering 4 out of 4 questions correctly. Experiment Results Predict Questions No Predict Questions Stop & Think Questions For home pregnancy tests You correctly answered: the antigen is human chorionic gonadotropin. 1 Which of the following is true of the indirect ELISA? You correctly answered: A positive patient sample contains antibodies speci fi c to the antigen of interest. 2 What does the antigen bind to on the antibody? You correctly answered: the variable region. 3 Which of the following is true of the direct ELISA? You correctly answered: The positive patient sample contains the speci fi c antigen. 4 What must occur in order for an HIV-infected individual to test positive with the indirect ELISA test? You correctly answered: seroconversion. 1

Experiment Data Sample Optical Density HIV Test Result Positive Control 1.624 + Negative Control 0.154 - Patient A 0.054 - Patient B 0.432 IND Patient C 1.990 + Post-lab Quiz Results You scored 100% by answering 4 out of 4 questions correctly. Review Sheet Results What is the key substance found in the developing bu ff er? You correctly answered: enzyme linked to the secondary antibody. 2 Antibodies are made of You correctly answered: proteins. 1 How would you describe the binding site of the secondary antibody? You correctly answered: the constant region of primary antibody. 2 A positive result for HIV in the indirect ELISA performed contains You correctly answered: antibodies against HIV. 3 Which patient needs to be retested? You correctly answered: patient B. 4 Describe how the direct and indirect ELISA are di ff erent. Your answer: In a direct Elisa you are detecting the presence of an antigen and the primary antibody used is the enzyme linked antibody. In an indirect Elisa you are detecting 1

the antibody, and the secondary antibody is enzyme linked. Discuss why a patient might test indeterminate. Your answer: A patient might test indeterminate if they have not yet seroconverted or has not yet produced enough antibodies to yield a positive result. 2 How would your results have been a ff ected if your negative control had given an indeterminate result? Your answer: If the negative control had given an indeterminate result, the results would have been invalidates due to not enough washing during the washing steps. 3 Brie ﬂ y describe the basic structure of antibodies. Your answer: Antibodies are made up of amino acids in the Y shape. This shape allows for two antigen binding sites. They also have a variable region that acts as the binding site for epitope. 4

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PhysioEx Exercise 12 Activity 3

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