Sayer Lelling

AP Bio

Immunology Virtual Lab

http://www.hhmi.org/biointeractive/vlabs/immunology/index.html

Answer IN YOUR OWN WORDS:

1. What is a false positive? How is it possible?
A false positive is when a specific antibody reacts with an antigen
nonspecifically. This is a false positive because it does not actually confirm if the
specific antibody is in the patient’s blood or not.
2. Why does the presence of an antibody to influenza virus indicate a person was exposed
to influenza?
The body produces antibodies when it encounters the antigen and is

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activated to do so. If the body was not exposed to influenza the body would have no

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use for the antibody and thus not produce it. Because the antibody is present we

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know the body had been exposed to influenza at some point in time.
3. What kind of condition is lupus (SLE)?

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Lupus (SLE) is believed to be an autoimmune disease, however the
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cause has yet to be determined.
4. How will the ELISA test help diagnose lupus?
The ELISA is an assay that measures the activity of enzymes that are
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linked to antibodies. By measuring the activity of the enzyme using ELISA we will
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learn how active the antibody is in the body, and thus whether the person has
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lupus.
5. Why do the blood samples need to be spun in the centrifuge?
The blood samples need to be spun in the centrifuge to spin out the
red blood cells and other blood components. This creates two layers with the
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components above and the serum below.

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6. What are serial dilutions and why are they used for this assay?
Serial dilutions are samples that are diluted in a systematic, organised
manner to determine the concentration of the sample or the sensitivity of the assay.
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7. Why has the ELISA plate been pretreated with the SLE antigen?
The ELISA plate has been treated with SLE antigen to eliminate bare
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spots that will allow the antibody to stick to the plastic of the wells.
8. What is the value of the positive control for this assay?
The positive control allows the experiment to account for the fact that
biological and chemical reagents used in ELISA could change over time. The positive
control should always produce positive results if done correctly.
9. Why is the buffer used as the negative control?
The buffer is used as a negative control because the SLE should never
react with the solution. If it does, then the patient’s cannot be trusted and the assay
should be repeated.
10. Why is each well washed in step 6?

https://www.coursehero.com/file/35392558/Immunology-Virtual-Labdocx/
 Washing removes any SLE antigen that did not react with the antigen
from the well. The antibody that has reacted with the antigen remains attached to
the well surface. If the antibody is not removed a false positive will likely occur.
11. Why are secondary antibodies added to each well (in step 7)?
The second antibody will react with the human antibody rather than
the antigen. Thus, it is used to detect the presence of SLE antibodies that may be
attached to the SLE antigens.
12. Why is the HRP-substrate added (in step 10)?
The HRP-substrate is added because it is the most commonly used
enzyme that will react with an antibody. The enzyme with hydrogen peroxide will
react with a chemical called ABTS and turn the solution yellow.
13. Explain how your results (the colored wells) indicate that patient A likely has SLE, patience
C likely does NOT have it, and patient B might have it.
If the wells are yellow, then the enzyme detected the SLE antibodies.

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Because the wells for patient A were yellow, it is likely that the patient has SLE.

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Because the wells for patient C were NOT yellow, the patient likely does not have

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SLE. Because SOME of the wells for patient B were yellow, it is unclear whether or
not the patient has SLE, and additional testing should be done.

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14. This virtual lab was testing for lupus. How is this same test used to test for the presence of
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HIV? If the results for an HIV test were the same as in this exercise, what would they indicate
about the three patients?

If this lab were testing for the presence of HIV the concept would be
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the same, except instead of SLE antigens, HIV antigens would be used. If the results
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were the same as in this experiment they would indicate the same thing. Patient A
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would likely have HIV, patient C would likely not, and further testing would be
required for patient B.
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https://www.coursehero.com/file/35392558/Immunology-Virtual-Labdocx/

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