Enzyme-linked immunosorbent assay (ELISA) - Animation

Dr.G Bhanu Prakash Animated Medical Videos
11 Feb 201901:22

Summary

TLDRELISA, or enzyme-linked immunosorbent assay, is a highly sensitive method for detecting specific antibodies or antigens. The Direct ELISA tests for antigen presence by adsorbing an unknown antibody to a microtiter plate, followed by the addition of a sample and an enzyme-linked antibody. If the antigen is present, it binds and retains the enzyme, and a color change indicates its presence. The Indirect ELISA involves adding antigen to the plate, then serum suspected of containing antibodies, and an enzyme-linked antibody that reacts with the constant region of antibodies. A color change after substrate addition confirms the antibody's presence, offering a reliable way to identify immune responses.

Takeaways

  • 🧪 ELISA is a highly sensitive test used to detect antibodies or specific antigens.
  • 🔍 The Direct ELISA is used to test for the presence of an antigen by adsorbing an unknown antibody to the well of a microtiter plate.
  • 💧 After rinsing to remove excess antibody, a sample suspected of containing the antigen is added to the well.
  • 🔗 An enzyme-linked antibody that can react with the antigen is added, binding to the antigen if present, and is retained in the well.
  • 🎨 The addition of a colorless substrate for the enzyme leads to color development, indicating the presence of the antigen.
  • 🔄 The Indirect ELISA involves adding antigen to the microtiter plate well, allowing it to attach to the walls.
  • 💧 Rinsing again removes excess antigen before adding serum suspected of containing antibodies.
  • 🔗 An enzyme-linked antibody that reacts with the constant region of other antibodies is added in the Indirect ELISA.
  • 🎨 The addition of a colorless substrate in the Indirect ELISA, followed by color development, indicates the presence of the identified antibody.
  • 🔬 Both Direct and Indirect ELISA methods rely on the specific binding of antibodies to antigens for detection.
  • 📈 The development of color in ELISA tests is a visual indicator of the presence of the target antigen or antibody, making it a qualitative and semi-quantitative assay.

Q & A

  • What is the enzyme-linked immunosorbent assay (ELISA)?

    -ELISA is an extremely sensitive test used to detect antibodies or specific antigens in a sample.

  • What is the purpose of the direct ELISA test?

    -The direct ELISA test is used to detect the presence of antigens by using an unknown antibody that is adsorbed to the well of a microtiter plate.

  • How is the antigen detected in a direct ELISA test?

    -In a direct ELISA, the antigen is detected by adding a sample suspected of containing the antigen. If the antigen is present, an enzyme-linked antibody that can react with the antigen is added and binds to it, which is then retained in the well.

  • What role does the enzyme-linked antibody play in a direct ELISA test?

    -The enzyme-linked antibody in a direct ELISA test binds to the antigen if it is present, indicating its presence through the retention in the well.

  • What is added after the enzyme-linked antibody in a direct ELISA test?

    -A colorless substrate for the enzyme is added after the enzyme-linked antibody in a direct ELISA test.

  • How is the presence of the antigen indicated in a direct ELISA test?

    -The presence of the antigen is indicated by the development of color after the addition of the colorless substrate for the enzyme.

  • What is the indirect ELISA test used for?

    -The indirect ELISA test is used to detect the presence of antibodies by adding antigen to the microtiter plate well and allowing it to attach to the walls.

  • How does the indirect ELISA test differ from the direct ELISA test?

    -In the indirect ELISA test, the antigen is added to the well first, followed by the serum suspected of containing antibodies, and then an enzyme-linked antibody that reacts with the constant region of other antibodies is added.

  • What is the final step in identifying the presence of antibodies in an indirect ELISA test?

    -The final step in an indirect ELISA test is the addition of a colorless substrate, and the development of color indicates the presence of the antibody being identified.

  • Why is rinsing necessary in both direct and indirect ELISA tests?

    -Rinsing is necessary to remove excess antibody or antigen, ensuring that only specific interactions are detected and avoiding false positives.

  • What does the development of color signify in both direct and indirect ELISA tests?

    -The development of color in both direct and indirect ELISA tests signifies a positive result, indicating the presence of the antigen or antibody being tested for.

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Étiquettes Connexes
ELISA TestAntibody DetectionAntigen PresenceImmunosorbent AssayDiagnostic ToolBiological ResearchMedical TestingColorimetric IndicatorMicrotiter PlateEnzyme-Linked Antibody
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