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Questions and Answers
What distinguishes ELISA from other serological tests?
What distinguishes ELISA from other serological tests?
Which of the following is NOT a primary binding test?
Which of the following is NOT a primary binding test?
What type of binding test includes both agglutination and neutralization tests?
What type of binding test includes both agglutination and neutralization tests?
Which enzyme is commonly used in ELISA for labeling antibodies?
Which enzyme is commonly used in ELISA for labeling antibodies?
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What is the primary purpose of conducting serological tests?
What is the primary purpose of conducting serological tests?
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What material are microtiter plates typically made from in ELISA tests?
What material are microtiter plates typically made from in ELISA tests?
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Which type of ELISA is performed in wells of a microtiter plate?
Which type of ELISA is performed in wells of a microtiter plate?
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Why is polystyrene used for the wells in ELISA plates?
Why is polystyrene used for the wells in ELISA plates?
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What is one characteristic of a Cell ELISA?
What is one characteristic of a Cell ELISA?
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Which of the following describes the primary function of the microtiter plate in ELISA?
Which of the following describes the primary function of the microtiter plate in ELISA?
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What is a significant advantage of IP staining over IF staining?
What is a significant advantage of IP staining over IF staining?
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Which type of microscope is needed for IP staining?
Which type of microscope is needed for IP staining?
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What characteristic of IP staining is emphasized in the content?
What characteristic of IP staining is emphasized in the content?
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Which statement is true regarding IP staining compared to IF staining?
Which statement is true regarding IP staining compared to IF staining?
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Which of the following is NOT an advantage of IP staining over IF staining?
Which of the following is NOT an advantage of IP staining over IF staining?
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What type of result does the ELISA method provide for antibody detection?
What type of result does the ELISA method provide for antibody detection?
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How is a qualitative result of an ELISA test identified?
How is a qualitative result of an ELISA test identified?
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Which statement about ELISA results is true?
Which statement about ELISA results is true?
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What is NOT a characteristic of the qualitative result in ELISA testing?
What is NOT a characteristic of the qualitative result in ELISA testing?
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Why is visual inspection important in ELISA testing for antibodies?
Why is visual inspection important in ELISA testing for antibodies?
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What is a characteristic of direct labeling of primary antibodies?
What is a characteristic of direct labeling of primary antibodies?
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What does indirect ELISA primarily focus on?
What does indirect ELISA primarily focus on?
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Which statement regarding indirect ELISA is true?
Which statement regarding indirect ELISA is true?
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What is a significant downside of direct labeling of primary antibodies?
What is a significant downside of direct labeling of primary antibodies?
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In the context of antibody detection, what does 'Ag' refer to?
In the context of antibody detection, what does 'Ag' refer to?
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What is one significant disadvantage of using radioimmunoassay (RIA)?
What is one significant disadvantage of using radioimmunoassay (RIA)?
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Which of the following is NOT an application of radioimmunoassay?
Which of the following is NOT an application of radioimmunoassay?
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Radioimmunoassay is primarily used for the measurement of which type of substances?
Radioimmunoassay is primarily used for the measurement of which type of substances?
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What is a common concern regarding the use of radioimmunoassay in medical settings?
What is a common concern regarding the use of radioimmunoassay in medical settings?
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Which characteristic primarily limits the use of radioimmunoassay?
Which characteristic primarily limits the use of radioimmunoassay?
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Study Notes
Serological Tests
- Serological tests are used to identify infectious diseases.
- These tests measure antigen-antibody reactions.
- Antigens are substances inducing an immune response.
- Antibodies are proteins produced by the immune system to counter a specific antigen.
Antigen
- Antigens are any substance that, when introduced into the body, stimulates the immune system to produce an immune response specific to that substance.
- This response is not directed at unrelated materials.
- Antigens have epitopes (antigenic determinants) which are the parts recognized by antibodies.
- Antibodies have paratopes which are the areas which recognize specific epitopes.
Antibody
- Antibodies are specific to microbes.
- Antibodies are also specific to species (in the same Ab).
- Antibodies have two important parts, the Fab region and the Fc region..
- Fab region binds to the antigen.
- Fc region controls the antibody function.
Antibody Labelling
- Antibodies can be labeled with enzymes or fluorescent dyes.
- The label is used to visualize the antigen-antibody complex.
- For example, enzyme labeling enables color change measurement.
- Fluorescent labeling allows visualization using a microscope.
Antigen-Antibody Reactions
- Serological reactions are used in diagnostic tests.
- Antigen-antibody reactions are used for diagnosing infectious diseases.
Serological Tests: Types
- Serological tests include primary, secondary and tertiary binding tests.
- ELISA (enzyme-linked immunosorbent assay), which includes Radioimmunoassay (RIA) is used for antigen/antibody detection and measuring.
- Different types of ELISA include Direct ELISA, Indirect ELISA and Sandwich ELISA.
- Direct ELISA: direct detection of antigen.
- Indirect ELISA: indirect detection of antibody.
- Sandwich ELISA: detection of antigen using two different antibodies.
- Immunofluorescence (IF) and immunoperoxidase (IP) are other types.
- Immunofluorescence (IF): uses fluorescent dyes for visualizing antigens.
- Immunoperoxidase (IP): uses enzymes for visualizing antigens.
ELISA Principle
- ELISA is a serological test determining antigen-antibody reactions.
- The reaction is detected by labeling the antibody with an enzyme (like horseradish peroxidase or alkaline phosphatase).
- Adding a substrate to the reaction creates a color change proportional to the amount of antigen or antibody.
- This color change is measured using a spectrophotometer.
ELISA: Types and Procedures
- ELISA can be direct or indirect.
- Direct ELISA involves coating wells with the antigen, adding the antibody (labeled with an enzyme), and detecting with the substrate .
- Indirect ELISA involves coating wells with the antigen, adding the serum to look for the presence of specific antibodies, then adding a secondary antibody specific to the first and detecting with the appropriate substrate.
- Sandwich ELISA involves coating the well with an antibody directed against the antigen, adding the antigen (which is being looked for), and a labeled secondary antibody directed against the antigen.
Immunofluorescence (IF)
- Used to identify antigens within cells.
- Antibodies with fluorescent tags are added to the sample.
- The fluorescence is observed under a microscope.
- Direct IF: detects antigen directly using labeled antibody.
- Indirect IF: detects antibodies indirectly using labeled secondary antibody.
- This method is useful in identifying the location and distribution of antigens in different tissue and cell types
Immunoperoxidase (IP)
- Identifies antigen locations using enzymes (peroxidases) or dyes linked to antibodies.
- Suitable for ordinary light microscopes to visualize stains.
- The method is very similar to the previous methods.
Advantages and Disadvantages of Different Tests
- Advantages and disadvantages of various techniques are covered in the presentation.
- These include cost, speed, sensitivity, and specificity
Reading ELISA Results
- Qualitative results are determined by naked-eye color change.
- Quantitative results are determined by color intensity measurements with a spectrophotometer.
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Description
This quiz covers the key concepts related to serological tests, including the roles of antigens and antibodies in the immune response. It explores how these components interact in identifying infectious diseases and highlights the structure of antibodies and their functions in targeting specific antigens.
