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CLS 1113 Introduction to Clinical Laboratory Practices

CLS 1113 Introduction to Clinical Laboratory Practices. Unit 5 Labeled Immunoassays Chapter 10. Labeled Immunoassays. Designed for Ags and Abs that DO NOT react in precipitation or agglutination tests due to their small size or low concentrations. Indirect method of detection:

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CLS 1113 Introduction to Clinical Laboratory Practices

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  1. CLS 1113Introduction to Clinical Laboratory Practices Unit 5 Labeled Immunoassays Chapter 10

  2. Labeled Immunoassays • Designed for Ags and Abs that DO NOT react in precipitation or agglutination tests due to their small size or low concentrations. • Indirect method of detection: • Competitive vs. Non-Competitive • Test Antigen or Antibody competes for binding sites

  3. Elements of Labeled Immunoassays • Ligands • Antibodies • Standards or Calibrators • Separation Methods • Detection of Label

  4. Radioimmunoassay (RIA) • Competitive binding assay • Uses a radioactive substance as a label • 3H - Tritiated hydrogen • 125I - Iodine 125

  5. Radioimmunoassay • Two Types • Number 1

  6. Radioimmunoassay Number 2

  7. Enzyme Immunoassay • Immunoassay labels • Enzymes • Cheap • Readily available • Long shelf life • Easily adapted to automation

  8. ELISA, Figure 10-4, page 149

  9. Enzyme Immunoassay • Enzymes are naturally occurring molecules that catalyze specific biochemical reactions. • They react with suitable substances to produce products that are chromogenic (color), fluorogenic, or luminescent.

  10. ELISA: Sandwich method Figure 10-5, page 149

  11. Fluorescent Immunoassay • Similar to ELISA but a fluorochrome is used rather than an enzyme. • Fluorochromes have the ability to absorb energy from light an emit it at a longer wavelength.

  12. Fluorescent Immunoassay: Direct and Indirect

  13. Chemiluminescent Immunoassays • Chemiluminescence is the production of light energy due to a chemical reaction. • Certain substances when oxidized can give off short or long bursts of light energy.

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