Podcast
Questions and Answers
Antibodies are produced by _______.
Antibodies are produced by _______.
plasma cells
The fluid portion of the blood with the clotting factors removed is called _______.
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 _______.
The form of Chlamydia that divides inside the host cell is the _______.
reticulate body
The direct fluorescent antibody test for Chlamydia uses fluorescently labeled _______.
The direct fluorescent antibody test for Chlamydia uses fluorescently labeled _______.
The washing steps are necessary to _______.
The washing steps are necessary to _______.
A positive result with the negative control would _______.
A positive result with the negative control would _______.
How many patient samples were positive for Chlamydia?
How many patient samples were positive for Chlamydia?
Antigens with epitopes in common _______.
Antigens with epitopes in common _______.
Which of the following describes the identity seen between human serum albumin and bovine serum albumin?
Which of the following describes the identity seen between human serum albumin and bovine serum albumin?
Which type of identity would you expect to see between human serum albumin and sheep serum albumin?
Which type of identity would you expect to see between human serum albumin and sheep serum albumin?
Which of the following describes the precipitin line formation between wells 2 and 3?
Which of the following describes the precipitin line formation between wells 2 and 3?
In the Ouchterlony technique, a precipitin line forms where _______.
In the Ouchterlony technique, a precipitin line forms where _______.
When two antigens have NO epitopes in common, their precipitin lines form _______.
When two antigens have NO epitopes in common, their precipitin lines form _______.
What is the medium for diffusion in the Ouchterlony technique?
What is the medium for diffusion in the Ouchterlony technique?
The antibodies used in this activity have been produced in _______.
The antibodies used in this activity have been produced in _______.
Possible reasons for an indeterminate test include _______.
Possible reasons for an indeterminate test include _______.
What result(s) would you expect if you had forgotten to add the developing buffer?
What result(s) would you expect if you had forgotten to add the developing buffer?
Why might a patient infected with HIV initially test negative with the indirect ELISA?
Why might a patient infected with HIV initially test negative with the indirect ELISA?
In the direct ELISA, _______.
In the direct ELISA, _______.
Within a class of antibodies, the _______.
Within a class of antibodies, the _______.
In the indirect ELISA, _______.
In the indirect ELISA, _______.
In both the direct and indirect ELISA, the substance that changes from colorless to colored in a positive result is the _______.
In both the direct and indirect ELISA, the substance that changes from colorless to colored in a positive result is the _______.
The proteins separated on the nitrocellulose in this activity are _______.
The proteins separated on the nitrocellulose in this activity are _______.
In this activity, _______________ was used to detect a positive result.
In this activity, _______________ was used to detect a positive result.
The sample(s) from which of the following tested positive for reverse transcriptase?
The sample(s) from which of the following tested positive for reverse transcriptase?
After the proteins are separated by electrophoresis, the _______.
After the proteins are separated by electrophoresis, the _______.
Western blotting _______.
Western blotting _______.
The ELISA is easier to perform and less expensive than the Western blot.
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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