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Questions and Answers
What principle does immunoassay auto analyzers primarily utilize?
What principle does immunoassay auto analyzers primarily utilize?
In a direct ELISA, which component is primarily coated on the microtiter well?
In a direct ELISA, which component is primarily coated on the microtiter well?
Which type of ELISA is specifically designed to detect antibodies in a sample?
Which type of ELISA is specifically designed to detect antibodies in a sample?
What happens to free primary antibodies during the indirect ELISA process?
What happens to free primary antibodies during the indirect ELISA process?
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What is the main purpose of adding a substrate in the ELISA process?
What is the main purpose of adding a substrate in the ELISA process?
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Which ELISA method requires both a primary and a secondary antibody for detection?
Which ELISA method requires both a primary and a secondary antibody for detection?
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The sandwich ELISA technique specifically helps to detect which type of molecule?
The sandwich ELISA technique specifically helps to detect which type of molecule?
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In a competitive ELISA, what is the primary reaction taking place?
In a competitive ELISA, what is the primary reaction taking place?
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Which of the following immunoassay techniques uses radioactivity?
Which of the following immunoassay techniques uses radioactivity?
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What type of substrates are typically used in ELISA for color development?
What type of substrates are typically used in ELISA for color development?
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What is the main function of the enzyme-linked secondary antibody in a Sandwich ELISA?
What is the main function of the enzyme-linked secondary antibody in a Sandwich ELISA?
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In a Competitive ELISA, what does a higher concentration of antigen in the sample indicate?
In a Competitive ELISA, what does a higher concentration of antigen in the sample indicate?
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Which of these is NOT a requirement for conducting an ELISA test?
Which of these is NOT a requirement for conducting an ELISA test?
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Which enzymes are most commonly used as labels in ELISA tests?
Which enzymes are most commonly used as labels in ELISA tests?
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What type of results can an ELISA test provide?
What type of results can an ELISA test provide?
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What is the primary purpose of the ELISA reader?
What is the primary purpose of the ELISA reader?
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Which component is responsible for forming a colored product in the ELISA process?
Which component is responsible for forming a colored product in the ELISA process?
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What function does washing serve in both the Sandwich and Competitive ELISA?
What function does washing serve in both the Sandwich and Competitive ELISA?
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Which among the following specimens can be tested using ELISA?
Which among the following specimens can be tested using ELISA?
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What condition must be met for an ELISA test to detect the presence of a specific antigen?
What condition must be met for an ELISA test to detect the presence of a specific antigen?
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Study Notes
Immunoassay Auto Analyzer
- Immunoassay is a biochemical test that determines the presence or concentration of a large molecule or small molecule in a solution using an antibody or antigen.
- An immunoassay auto analyzer is a lab instrument that automates many manual immunoassay techniques.
ELISA Technique
- ELISA stands for enzyme-linked immunosorbent assay.
- It's a plate-based technique used to detect and measure soluble substances like peptides, proteins, antibodies, and hormones.
- The technique relies on the highly specific interaction between antigens and antibodies.
Types of ELISA Assays
- ELISA tests are categorized based on the antigen-antibody binding methods:
- Direct ELISA: Antibody is coated onto the microtiter well.
- Indirect ELISA: Antigen is coated onto the well, followed by antibody binding.
- Sandwich ELISA: Primary antibody is coated, then antigen binds, followed by a secondary antibody binding to the antigen.
- Competitive ELISA: Antigen competes with labeled antigen to bind to the antibody coated on the well.
Direct ELISA
- This Sandwich ELISA variant detects antigens in a sample.
- The microtiter well is coated with antibody.
- Sample containing antigen is added and unbound antigen is removed.
- Enzyme-specific substrate is added, yielding a measurable colored product.
Indirect ELISA
- Indirect ELISA identifies antibodies in a sample.
- Antigen is coated onto the microtiter well.
- Sample containing antibodies is added to bind to the antigen.
- Enzyme-conjugated secondary antibody is used to detect the antigen-antibody complex.
- Specific substrate creates a measurable colored product.
Sandwich ELISA
- This ELISA type detects antigens through a dual antibody capture.
- A coated antibody on the well binds the antigen.
- An enzyme-linked secondary antibody binds to another epitope on the antigen.
- The enzyme substrate produces a measurable colored product
Competitive ELISA
- This ELISA detects antigen concentration by competition.
- The microtitre well is coated with antigen.
- Sample containing antibodies and antigen incubate together.
- The amount of free antibody is inversely correlated to the concentration of antigen in the sample.
ELISA Test Requirements
- Purified antigen or antibody
- Standard solutions (positive and negative controls)
- Sample to be tested
- Microwell plates
- Wash buffer
- Enzyme-labeled antibody and substrate
Enzymes used in ELISA
- Horseradish peroxidase (HRP) and alkaline phosphatase (AP) are commonly used.
ELISA Reader Components
- ELISA reader is a sophisticated instrument to detect proteins in a liquid sample.
- The system components include:
- ELISA plate
- Light source
- Optical system
- Optic fiber
ELISA Results and Interpretation
- ELISA result is expressed with OD450 values and a correlation is used to quantify the amount of antigen or antibody. The concentration of antigen in the sample is shown in a graph.
- The value is plotted against the known concentrations of positive and negative solutions.
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Description
Explore the basics of immunoassays and the ELISA technique through this quiz. Learn about the different types of ELISA assays and how immunoassay auto analyzers enhance laboratory efficiency. Test your knowledge on these essential biochemical testing methods.
