Questions and Answers
Antibodies are produced by _______.
plasma cells
The fluid portion of the blood with the clotting factors removed is called _______.
serum
The form of Chlamydia that divides inside the host cell is the _______.
reticulate body
The direct fluorescent antibody test for Chlamydia uses fluorescently labeled _______.
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The washing steps are necessary to _______.
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A positive result with the negative control would _______.
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How many patient samples were positive for Chlamydia?
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Antigens with epitopes in common _______.
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Which of the following describes the identity seen between human serum albumin and bovine serum albumin?
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Which type of identity would you expect to see between human serum albumin and sheep serum albumin?
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Which of the following describes the precipitin line formation between wells 2 and 3?
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In the Ouchterlony technique, a precipitin line forms where _______.
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When two antigens have NO epitopes in common, their precipitin lines form _______.
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What is the medium for diffusion in the Ouchterlony technique?
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The antibodies used in this activity have been produced in _______.
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Possible reasons for an indeterminate test include _______.
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What result(s) would you expect if you had forgotten to add the developing buffer?
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Why might a patient infected with HIV initially test negative with the indirect ELISA?
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In the direct ELISA, _______.
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Within a class of antibodies, the _______.
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In the indirect ELISA, _______.
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In both the direct and indirect ELISA, the substance that changes from colorless to colored in a positive result is the _______.
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The proteins separated on the nitrocellulose in this activity are _______.
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In this activity, _______________ was used to detect a positive result.
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The sample(s) from which of the following tested positive for reverse transcriptase?
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After the proteins are separated by electrophoresis, the _______.
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Western blotting _______.
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The ELISA is easier to perform and less expensive than the Western blot.
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Study Notes
Antibodies and Blood Components
- Antibodies are produced by plasma cells.
- Serum is the fluid portion of blood after clotting factors are removed.
Chlamydia and Antigen Detection
- The reticulate body is the form of Chlamydia that divides inside host cells.
- Direct fluorescent antibody tests utilize fluorescently labeled antibodies to detect specific antigens in patient samples.
- Washing steps in tests are vital to decrease nonspecific binding.
Testing Results and Interpretations
- A positive result with a negative control leads to a "false positive," rendering the results invalid.
- Only one patient sample tested positive for Chlamydia.
- Antigens that share epitopes can exhibit partial identity or be identical.
Serum Albumin Identifications
- Human serum albumin shows partial identity when compared to bovine serum albumin and sheep serum albumin.
- Precipitin lines between wells indicate antigen-antibody interaction; an arc signifies partial identity whereas an X indicates no shared epitopes.
Ouchterlony Technique
- Optimal antigen-antibody ratio forms a precipitin line in the Ouchterlony test medium, which is agar.
Indeterminate Test Results
- Indeterminate test results may occur due to low antibody levels, recent infection, or ineffective washing steps.
- Forgetting to add the developing buffer can cause false negatives for positive controls and patient samples.
ELISA Techniques
- Indirect ELISA could yield negative results in HIV patients still undergoing seroconversion.
- Direct ELISA involves both primary and secondary antibodies binding to the antigen.
- In indirect ELISA, the secondary antibody attaches to the primary antibody's constant region.
Substrates and Controls
- In ELISAs, substrates change from colorless to colored in positive results.
- Proteins separated in this lab are HIV antigens, detected by an indirect ELISA.
Western Blotting Techniques
- Proteins are transferred to a nitrocellulose membrane and treated with antibodies post-electrophoresis.
- Western blotting is acknowledged as a serological technique.
- ELISA is often preferred over Western blot due to its simplicity and cost-effectiveness.
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