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Exercise 12: Serological Testing: Activity 1: Using Direct Fluorescent Antibody Technique to Test for Chlamydia Lab Report

Pre-lab Quiz Results


You scored 100% by answering 4 out of 4 questions correctly.
1. Which of the following has an epitope? You
correctly answered: b. an antigen

2. Chlamydia trachomatis is
You correctly answered: c. a bacterium that reproduces inside its host cell.
3. The infectious form of Chlamydia that is spread from person to person is the You
correctly answered: d. elementary body that stains green in this activity

4. The goal of the direct fluorescent antibody test is to test for the presence of You
correctly answered: c. antigen.

Experiment Results
Stop & Think Questions:
The source of the patient samples is
You correctly answered: c. epithelial scrapings
What is the name for the specific part of the antigen that the antibody binds to?
You correctly answered: b. epitope

15. Drag the first slide (patient A) to the fluorescent microscope.


Count the number of elementary bodies you see through the microscope (recall that elementary bodies stain green), enter
the number of elementary bodies in the field below, and then click Submit Data to display your results in the grid. After you
click Submit Data, the slide will automatically be placed in the biohazardous waste disposal.
You answered: 0 elementary bodies
16. Drag the next slide (patient B) to the fluorescent microscope.
Count the number of elementary bodies you see through the microscope (recall that elementary bodies stain green), enter
the number of elementary bodies in the field below, and then click Submit Data to display your results in the grid. After you
click Submit Data, the slide will automatically be placed in the biohazardous waste disposal.
You answered: 15 elementary bodies
17. Drag the next slide (patient C) to the fluorescent microscope.
Count the number of elementary bodies you see through the microscope (recall that elementary bodies stain green), enter
the number of elementary bodies in the field below, and then click Submit Data to display your results in the grid. After you
click Submit Data, the slide will automatically be placed in the biohazardous waste disposal.
You answered: 1 elementary bodies
18. Drag the next slide (positive control) to the fluorescent microscope.
Count the number of elementary bodies you see through the microscope (recall that elementary bodies stain green), enter
the number of elementary bodies in the field below, and then click Submit Data to display your results in the grid. After you
click Submit Data, the slide will automatically be placed in the biohazardous waste disposal.
You answered: 19 elementary bodies
19. Drag the last slide (negative control) to the fluorescent microscope.

Count the number of elementary bodies you see through the microscope (recall that elementary bodies stain green), enter
the number of elementary bodies in the field below, and then click Submit Data to display your results in the grid. After you
click Submit Data, the slide will automatically be placed in the biohazardous waste disposal.
You answered: 0 elementary bodies Experiment

Data:
Sample
Patient A
Patient B

Number of Elementary Bodies


0
15

Chlamydia Result
Negative
Positive

Sample
Patient C
Positive Control
Negative Control

Number of Elementary Bodies


1
19
0

Chlamydia Result
Positive
Positive
Negative

Post-lab Quiz Results


You scored 100% by answering 5 out of 5 questions correctly.
1. In the serology test just performed, the fluorescence is found on the You
correctly answered: c. antibody

2. What would most likely happen if you forgot to fix the samples to the slide?
You correctly answered: a. They would all test negative because the washing steps would remove the samples.
3. Which sample showed some residual nonspecific binding after the washing steps? You
correctly answered: c. patient C

4. Which patient samples contained the Chlamydia-specific antigen? You


correctly answered: b. patient B

5. How is chlamydia disease transmission prevented?


You correctly answered: d. abstinence or faithful, mutual monogamy

Review Sheet Results


1. Describe the importance of the washing steps in the direct antibody fluorescence test.
Your answer:
It is very important to following the washing steps in the direct antibody fluorescence test in order to remove any non specific
binding that may have occurred.

2. Explain where the epitope (antigenic determinant) is located.


Your answer:
The epitopes are located in the antigens so that the antibodies can bind to them.
3. Describe how a positive result is detected in this serological test. Your
answer:
The elementary bodies of the Chlamydia trachomatis stains green inside the red host cell, and the presence of ten or more
elementary bodies in a field of view with a diameter of 5mm is considered a positive result.

4. How would the results be affected if a negative control gave a positive result? Your
answer:
This would be a false positive which would invalidate all other results.

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