Name: Sara BolandLab: Saturday 8:00 a.m.

Lab Name: ELISA Simulation Lab

Purpose: The body fights infection from pathogens in many ways. One of which is through the development the Specific Immune Response. Specific Immune Responses are tailored to the specific pathogen invading the body. A specific immune response is triggered by proteins and other molecules produced by pathogens. These are called antigens. The T and B cells in the immune system are stimulated to reproduce and generate antibodies to fight infection once they are alerted to the presence of antigens. Phagocytic macrophages and dendritic break down pathogens and display the antigenic molecules so that they can be recognized by lymphocytes. Antibodies are immunoglobulins that bind to antigens and mark them for destruction. The indirect ELISA is as test that looks for pathogenic infection by recognizing the presence of antibodies in the blood. ELISA stands for enzyme-linked immunoabsorbant assay and the purpose of this lab is to use this multi-step assay to test samples for infection of a specific pathogen, in this case Simulated Avian Flu, which is an extremely dangerous influenza virus caused by H5N1.

Materials:-Plastic pipettes -Microtiter plate-Six patient samples: A, B, C, D, E, F -Simulated secondary antibody-Simulated chromogen -Positive control-Negative control

Organisms:-Simulated Avian Flu

Procedure:*Note: a clean pipet is used for every step in the following procedure1. Using one pipet, administer 3 drops of Simulated Avian Flu in each well of rows A and B in the microtiter plate.2. Add 3 drops of positive control to wells A1, A2 and A33. Add 3 drops of negative control to wells A4, A5 and A64. Add 3 drops of Patient A sample to wells A7, A8 and A95. Add 3 drops of Patient B sample to wells A10, A11 and A126. Add 3 drops of Patient C sample to wells B1, B2 and B37. Add 3 drops of Patient D sample to wells B4, B5 and B68. Add 3 drops of Patient E sample to wells B7, B8 and B99. Add 3 drops of Patient F sample to wells, B10, B11 and B12

10. Add 3 drops of simulated secondary antibody to each well in rows A and B of the microtiter plate11. Add 3 drops of simulated chromogen to each well in rows A and B of the microtiter plate12. Incubate the microtiter plate at room temperature for between 5 and 10 minutes

Results:

Sample Color Test Resultpositive control dark purple positivenegative control light green negativePatient APatient BPatient CPatient DPatient EPatient F