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Questions and Answers
What is the role of a secondary antibody in an ELISA test?
What is the role of a secondary antibody in an ELISA test?
- To detect the presence of antigens directly
- To recognize the first antibody and facilitate colorimetric detection (correct)
- To enhance the immune response against foreign substances
- To serve as a negative control in the experiment
Which of the following best describes the appearance of positive results in a Biolog System test?
Which of the following best describes the appearance of positive results in a Biolog System test?
- A purple color resulting from the reduction of tetrazolium dye (correct)
- A blue hue indicating high acidic pH levels
- A colorless solution indicating no bacterial growth
- A green color change suggesting the presence of amino acids
What is the significance of using both positive and negative control groups in an ELISA test?
What is the significance of using both positive and negative control groups in an ELISA test?
- To ensure accurate colorimetric detection without any antibodies
- To increase the sensitivity of the test towards antigens
- To validate that the manipulation of variables affects the test results (correct)
- To represent different types of antibodies that might be present
In the Biolog System, what is used to measure the concentration of bacteria in suspension?
In the Biolog System, what is used to measure the concentration of bacteria in suspension?
What happens if too much bacteria is inoculated in a Biolog test?
What happens if too much bacteria is inoculated in a Biolog test?
What is the primary purpose of the washing steps in an ELISA test?
What is the primary purpose of the washing steps in an ELISA test?
What is indicated by a negative control group in an ELISA test?
What is indicated by a negative control group in an ELISA test?
During the ELISA test, what is the role of the substrate added at the end of the process?
During the ELISA test, what is the role of the substrate added at the end of the process?
What is the incubation temperature during the Biolog System test?
What is the incubation temperature during the Biolog System test?
How is colorimetric detection achieved in an ELISA test?
How is colorimetric detection achieved in an ELISA test?
Flashcards
Antigen
Antigen
A foreign substance that triggers an immune response.
Antibody
Antibody
A protein produced by the immune system to neutralize an antigen.
ELISA Test
ELISA Test
A lab test used to detect antigens or antibodies in a sample, like blood.
Secondary Antibody
Secondary Antibody
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Colorimetric Detection
Colorimetric Detection
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Positive Control (ELISA)
Positive Control (ELISA)
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Negative Control (ELISA)
Negative Control (ELISA)
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Biolog System
Biolog System
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Tetrazolium Dye
Tetrazolium Dye
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Turbidimeter
Turbidimeter
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Study Notes
ELISA Test
- Antigens: Foreign substances triggering an immune response.
- Antibodies: Proteins the body produces to neutralize antigens.
- ELISA (Enzyme-Linked Immunosorbent Assay): Used to detect antigens and antibodies.
- Applications: Diagnosing HIV, EBV, pregnancy, drug screening, food allergies.
- Secondary Antibody: Binds to the primary antibody, facilitating detection.
- Colorimetric Detection: Visual color change signals the presence of a serum antibody—a sign of exposure to a pathogen.
- Washing steps: Crucial to remove unbound reagents/materials. Prevents inaccurate results.
- Controls: Positive and negative controls needed to validate test accuracy.
- Positive control: Shows a positive result as antibodies are provided.
- Negative control: Shows no color change due to lack of antibodies.
- Enzyme Conjugation: Secondary antibody is conjugated with an enzyme for color detection.
ELISA Procedure
- Label wells.
- Add viral antigen to wells.
- Incubate at room temperature for 5 minutes.
- Remove liquid.
- Wash with PBS.
- Add test reagent to each well.
- Incubate at 37°C.
- Remove and wash.
- Add secondary antibody.
- Incubate at 37°C for 15 minutes.
- Rinse twice with PBS.
- Add substrate.
- Incubate.
- Observe wells for color change (positive result).
Biolog System
- Purpose: Identifying bacteria based on metabolic properties.
- Applications: Identifying gram-positive and gram-negative species.
- Method: Utilizes a 96-well plate.
- Carbon Sources: Sugars, amino acids, and carboxylic acids.
- Sensitivity Tests: Antibiotics, salt, and pH tolerance.
- Positive Result: Tetrazolium dye reduction leading to a purple color.
- Tetrazolium: A redox dye.
- Incubation Temperature: 33°C.
- Indicator: Bacteria growth indicated by purple color (dye reduction).
- Turbidimeter: Measures light transmission to estimate bacterial concentration in a suspension.
- Transmittance: Percentage of light passing through a sample.
- Negative Controls: Essential for bias detection.
Biolog System Procedure
- Unwrap and label the microplate.
- Collect bacterial sample from agar plate.
- Prepare bacterial suspension in inoculating fluid.
- Mix thoroughly.
- Determine bacterial transmittance using turbidimeter.
- Transfer entire bacterial suspension into a reservoir.
- Transfer bacterial suspension into each well.
- Incubate at 33°C.
- Observe for color development (3-36 hours).
- Interpret results.
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