- to demonstrate principles of
immunodiagnosis including how antibody-antigen interactions can be used
to screen for disease.
- to understand the clinical
features and transmission of Lyme disease including the life cycle of Ixodes,
the tick that transmits the spirochete B. burgdorferi
to humans.
Students
will:
- perform an enzyme linked
immunosorbent assay (ELISA) simulation for Lyme disease
- be introduced to the complexity
of assay development, the importance of controls and the meaning of
false negative and false positive results.
2.5 hours
This lab aligns with the
following Maine
performance indicators: A4, C4, D3, J1-3, K1-6, L1-3 and 8,
M1-4 and is allied with Advanced Placement Biology Lab #1 and 2
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Hands-on
- Use pipettors
- Bind antigens to microplate
- Make serial dilutions of antibody
- Complete assay/review results
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Lecture
- Lyme disease case discussion
- Antibody-antigen interactions
- Introduction to ELISA technology
- Discussion of titration and its uses
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A microtiter plate coated with the antigen will be provided to the students. The students will titer serum from a "patient" with possible Lyme disease and serum from a normal control. Enzyme labeled goat species specific anti-IgG will be added. The final step is the addition of a chromogen. If antibody has bound to the antigen, a color will appear. The concept of titer, antibody specificity, and the complexity of assay development are demonstrated in the unit.
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