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Questions and Answers
In single radial immunodiffusion, what is the role of the agar medium?
In single radial immunodiffusion, what is the role of the agar medium?
- It provides a semisolid matrix in which antibodies are embedded to react with diffusing antigens. (correct)
- It contains the antigens that diffuse outward to react with antibodies.
- It is used to label antibodies with fluorescent dyes for visualization.
- It serves as a surface for enzymatic reactions in ELISA.
What is the fundamental difference between direct and indirect immunofluorescence (IF) in terms of what they detect?
What is the fundamental difference between direct and indirect immunofluorescence (IF) in terms of what they detect?
- Direct IF is qualitative, while indirect IF is quantitative.
- Direct IF uses a secondary antibody, while indirect IF uses a primary antibody.
- Direct IF detects antigen, and indirect IF detects antibodies. (correct)
- Direct IF detects complement components, while indirect IF detects immunoglobulins.
In the context of precipitation tests, which term describes the soluble antigen and the antibody involved?
In the context of precipitation tests, which term describes the soluble antigen and the antibody involved?
- Enzyme and substrate
- Precipitinogen and precipitin (correct)
- Agglutinogen and agglutinin
- Fluorochrome and antibody
What is the primary advantage of using ELISA over other serological tests like immunodiffusion?
What is the primary advantage of using ELISA over other serological tests like immunodiffusion?
In diagnosing Systemic Lupus Erythematosus (SLE), which immunological technique is most commonly used to detect anti-nuclear antibodies (ANA)?
In diagnosing Systemic Lupus Erythematosus (SLE), which immunological technique is most commonly used to detect anti-nuclear antibodies (ANA)?
Which of the following methods is best suited for quantifying specific immunoglobulin levels in a patient's serum?
Which of the following methods is best suited for quantifying specific immunoglobulin levels in a patient's serum?
Which of the following is measured to assess cellular immunity?
Which of the following is measured to assess cellular immunity?
A lab technician is tasked with identifying a specific viral antigen in a patient sample using immunofluorescence. Which method should they employ for direct detection?
A lab technician is tasked with identifying a specific viral antigen in a patient sample using immunofluorescence. Which method should they employ for direct detection?
In a double immunodiffusion assay, what does the formation of a precipitation line between the antigen and antibody wells indicate?
In a double immunodiffusion assay, what does the formation of a precipitation line between the antigen and antibody wells indicate?
What is the purpose of using fluorochrome-labeled antibodies in flow cytometry?
What is the purpose of using fluorochrome-labeled antibodies in flow cytometry?
Flashcards
Precipitation (Immunodiffusion)
Precipitation (Immunodiffusion)
Antigen-antibody reaction with a soluble antigen, where the antigen is a precipitinogen and the antibody is a precipitin.
Clinical applications of Single (Radial) Immunodiffusion
Clinical applications of Single (Radial) Immunodiffusion
Quantitative assay of immunoglobulins, complement components (C1, C2, C3,..).
Double Immunodiffusion
Double Immunodiffusion
Antigen and antibody diffuse against each other in agar, forming a precipitate line where they meet; used for diagnosing bacterial, viral, or fungal diseases.
Immunofluorescent (IF) Microscopy
Immunofluorescent (IF) Microscopy
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Direct IF
Direct IF
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Indirect IF
Indirect IF
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Enzyme-Linked Immunosorbent Assay (ELISA)
Enzyme-Linked Immunosorbent Assay (ELISA)
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Clinical application of ELISA: Diagnosis of infectious diseases
Clinical application of ELISA: Diagnosis of infectious diseases
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Assessment of Cellular Immunity
Assessment of Cellular Immunity
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Flow Cytometry
Flow Cytometry
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Study Notes
- These notes discuss methods for measuring immune function, including precipitation tests, immunofluorescent microscopy, ELISA, and assessing cellular immunity.
Precipitation (Immunodiffusion)
- Precipitation reactions involve the reaction of soluble antigens with antibodies.
- In precipitation, the antigen is called precipitinogen and the antibody is called precipitin.
- Types of precipitation tests include the Ring test and Agar diffusion.
Single (Radial) Immunodiffusion
- Soluble antigens react with antibodies in a semisolid medium.
- This leads to the formation of precipitation lines.
- Used for quantitative assay of immunoglobulins.
- Useful for quantitative analysis of complement components like C1, C2, and C3.
Double Immunodiffusion
- Antigens and antibodies diffuse against each other in semisolid agar.
- Lines of precipitate form where antigen and antibody meet.
- Used for diagnosing various bacterial, viral, and fungal diseases.
Immunofluorescent (IF) Microscopy
- Involves an antigen-antibody reaction where antibodies are labeled with a fluorescent dye.
- The antigen-antibody complex is visualized using a fluorescence microscope.
- There are two main types of IF: direct and indirect.
Direct IF
- Used to detect antigens like viral, parasitic, intracellular bacteria (chlamydia), and tumor antigens.
- This is done using a specific fluorescein-labeled antibody.
Indirect IF
- Used to detect antibodies in patient serum.
- Secondary anti-human globulin conjugated with fluorescein is utilized.
- Applied to detect autoantibodies, such as anti-nuclear antibodies (ANA), in patients with systemic lupus erythematosus (SLE).
Enzyme-Linked Immunosorbent Assay (ELISA)
- ELISA can demonstrate either an unknown antigen or an unknown antibody.
- Serves as a quantitative assay and is highly sensitive.
- Used in the diagnosis of infectious diseases, including viral (HIV, HBV, HCV) and bacterial infections (Brucellosis, Syphilis)
- Aids in detecting hormones.
Direct ELISA
- Used to detect antigens.
- Involves adding an enzyme-conjugated secondary antibody after the antigen binds to the antibody-coated well.
- Substrate is added for color measurement
Indirect ELISA
- Used to detect antibodies such as HIV and HCV.
- A specific antibody is added to antigen-coated wells.
- Followed by an enzyme-conjugated secondary antibody and substrate for color measurement.
Assessment of Cellular Immunity
- Cellular immunity is measured by evaluating: phagocyte function and lymphocyte proliferation.
- Flow cytometry is used, where leukocytes are stained with fluorochrome-labeled antibodies.
- Aids in calculating the CD4/CD8 ratio in HIV follow-up.
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Description
Overview of methods to measure immune function, including precipitation tests, immunofluorescent microscopy, and ELISA. Covers precipitation reactions, single and double immunodiffusion. Discusses using these methods for diagnosing diseases.
