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Ab indicates_____, Ag indicates_____
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Serology is a measurement of...
Ag-Ab interactions
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Uses of Immunological Tests (10):
- Detect exposure
- Quantify Ab or Ag
- ID infectious agents
- Monitor exposure
- Vx titers
- Monitor proteins, hormones & drug levels
- Type blood & Tissue
- Recheck leukemia/lymphoma therapy
- ID tumor Ag & Auto-Ab
- ID cells in immune response
- Survey a population
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What is the basis for a variety of immunologic tests?
Ag-Ab interaction
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What affects the Ag-Ab interaction?
- Forces of attraction
- Concentration of Ag-Ab
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Titration-
Serum dilutions used to measure the amount of Ab in serum
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Purpose of a titer
Quantify the amount of Ab
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What methods can be used to detect Ab for a titration?
- Precipitation/Agglutination
- ELISA
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Using an ELISA, what would indicate a high titer?
+wells with color change
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Which value indicates a higher Ab concentration, 1:4 or 1:512?
1:512 = higher Ab concentration
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Two general types of Immunodiagnostic Tests:
- Primary Binding Tests
- Secondary Binding Tests
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Primary Binding Tests are used to....
Directly measure binding of Ag to Ab in vitro
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Secondary binding tests are used to....
Measure the RESULTS of Ag-Ab interactions in vitro
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Samples that can be used for in vitro tests (4):
- Serum
- Peritoneal Fluid
- Pleural Fluid
- CSF
- Synovial Fluid
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Cells/Tissues to probe with Ab when looking for Ag
- Whole blood in anticoag (RBC, WBC and Platelets)
- Tissue Sections on dead or euthanized animals or from biopsy
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What Reagents can be used in immunodiagnostic tests(5)?
- Complement
- Polyclonal Ab
- Monoclonal Ab
- Anti-globulins (Ab to Ab)
- Anti-serum
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Anti-Serum contains...
Ab against specific Ag
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What must ALWAYS be done before running a binding test?
Run a known positive and negative controls
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Two Major Types of Primary Binding Tests:
- Immunofluorescence Assays
- Immunoenzyme Assays
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Immunofluoroescence Assays include (2):
- Direct Fluorescent Ab Tests
- Indirect Fluorescent Ab Tests
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What do DFA and IFA detect?
- Ab
- (Direct/Indirect Fluorescent Ab)
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Immunoenzyme Assays include (3):
- ELISA
- Western Blot
- Immunohistochemistry
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Secondary Binding Tests include (4):
- Precipitation Tests
- Agglutination Tests
- Neutralization Tests
- Complement Fixation Tests
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What is the unknown in Direct Flourescent Ab Tests? Indirect FA?
- Direct= Ag
- Indirect= Serum
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Fluorochrome commonly used to label Ab?
FITC
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ELISA tests use wells coated with ___
Ag
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Indication of a Positive ELISA test?
Color Change
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Which ELISA test can be used for a titer?
ELISA
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Which test requires electrophoresis of proteins?
Western Blot
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Western Blot is probed with:
- Labeled Ab (protein Ag)
- P's serum (Ab)
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Controls used for an Immunohistochemistry:
- Irrelevant Ab
- Normal Tissue Sample
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In a Precipitation Test, what does the precipitate consist of?
Complex of Ab & SOLUBLE Ag
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