ELISA and Kirby Bauer Disk Diffusion Tests PDF
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Summary
This document provides information about ELISA and the Kirby Bauer disk diffusion test, including procedures, applications, and factors influencing results. It covers topics like innate resistance and antibiotic susceptibility testing. The document appears to be lab examination materials for an undergraduate-level microbiology course.
Full Transcript
odule 9 M ELISA steps to detect antigens or antibodies. The function of each step in the ELISA procedure. The applications of ELISA:to diagnose disease, biotech: to quantify antigens, production of vaccines (trigger antibody response) LISA: E Direct Sandwich Elisa de...
odule 9 M ELISA steps to detect antigens or antibodies. The function of each step in the ELISA procedure. The applications of ELISA:to diagnose disease, biotech: to quantify antigens, production of vaccines (trigger antibody response) LISA: E Direct Sandwich Elisa detects or quantifies an antigen. 1. Immobilization of capture antibody (attached to plate) 2. Wash 3. Blocking agent to seal plastic surface and prevent non-specific binding of subsequent reagents** 4. Wash 5. Add sample and controls (may or may not have the antigen) 6. Wash 7. Detection antibody (conjugated enzyme) 8. Wash 9. Add substrate 10.Production of colored or non-colored product (if antigen is present) *may get false positives if you do not wash between 6-8. Indirect ELISA detects or quantifies antibodies against a particular antigen. 1. Immobilize the antigen. 2. Wash 3. Blocking agent to seal plastic surface and prevent nonspecific binding of subsequent reagents 4. Waash 5. Add sample and controls (plasma or serum that may or may not have antibodies) 6. Wash 7. Detection antibody (conjugated enzyme)secondary antibody 8. Wash 9. Add substrate 10.Production/non-production of colored product (if antibody is present) odule 10 M Be familiar with the Kirby Bauer disk diffusion test and the factors that may interfere with your results. - Always use same medium, incubation time, and temperature - use aseptic techniques to prevent contamination - Microbial culture is pure - Correct amount of bacterial inoculum - Correct amount of antibiotics (not expired) - Proper inoculation Interpret the results of the Kirby Bauer disk diffusion test - Innate resistance with no halo (-) - If halo is higher >= 18 it is susceptible,
