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Precipitation
Combine soluble antigen and soluble antibody to produce insoluble complexes that are visible.
Agglutination
Particulate antigens form visible aggregates when an antibody is present.
Antigen-Antibody Binding Affinity
Initial attraction force between a Fab site on an antibody molecule and an epitope of determinant site on an antigen.
Antigen-Antibody Binding: Avidity
Sum of the attractive forces between an antigen and an antibody that keeps the molecule together.
Prozone phenomenon
Antibody excess where antigens combine with only one or two antibody molecules, resulting in a false negative.
Postzone phenomenon
Antigen excess where small aggregates are surrounded by excess antigen, resulting in a negative test result.
Zone of Equivalence
Point where the number of multivalent sites of antigen and antibody are approximately equal.
Radial Immunodiffusion
Single diffusion technique where antigen diffuses out until the point of equivalence is reached.
Ouchterlony Diffusion
Double diffusion method where antigen and antibody diffuse out radially to form a line of precipitation.
Identity (Ouchterlony Diffusion)
Antigens placed in the wells are identical.
Partial identity (Ouchterlony Diffusion)
Antigens in the wells are similar but not identical.
Non-identity (Ouchterlony Diffusion)
Antigens in the wells do not share any similarity.
Immunofixation Electrophoresis
Double-diffusion technique where unknown antigen is electrophoresed, and then antibody is applied directly to the gel.
Nephelometry
Technique measuring light scattered at an angle to indicate the amount of antigen or antibody present.
Radial Immunodiffusion (RID)
Technique where the square of the diameter of the ring of precipitation is proportional to the concentration.
Ouchterlony Diffusion Patterns
Concept to match the precipitation pattern with the correct interpretation.
Immuno-electrophoresis
Differentiation of serum proteins by electrophoresis followed by diffusion of antibody from the wells.
Immunofixation electrophoresis
Technique used for identifying over- or underproduction of antibody by electrophoresis of serum followed by direct application of antibody to the gel.
Direct Agglutination
Agglutination reaction using known bacterial antigens to test for unknown antibodies, such as in the Widal Test for Typhoid Fever.
Hemagglutination
Agglutination involving red blood cells as particles with antigens, commonly used in ABO blood group typing.
Passive agglutination
Agglutination method using particles coated with antigens not usually found on their surface, like erythrocytes or latex particles, to detect patient factors or antibodies.
Reverse Passive Agglutination
Agglutination technique employing antibodies attached to a carrier particle to detect microbial antigens, used for rapid identification of infectious agents.
Agglutination Inhibition
Agglutination reaction based on competition between particulate and soluble antigens for limited antibody-combining sites, used to detect antibodies to viruses like Rubella and Influenza.
Competitive Immunoassays
Immunoassays where the labeled antigen competes with unlabeled patient antigen for a limited number of antibody binding sites, with the bound label inversely proportional to the labeled antigen concentration.
Noncompetitive Immunoassays
Immunoassays where the antibody is first passively absorbed to a solid phase, allowing the unknown patient antigen to react and be captured by the antibody, with the measured label amount directly proportional to the patient antigen.
Homogenous Immunoassays
Does not require a separation or washing step to separate the bound from the free reactants
Heterogeneous Immunoassays
Often performed utilizing a solid phase for the reaction and washing away of unbound reactant
Radioimmunoassay's (RIA)
Uses radioactive labels (125 I most popular) and emits gamma radiation detected by a gamma counter; measures trace amounts of analytes like hormones, serum proteins, and vitamins
Enzyme Immunoassays (EIA)
Uses enzymes as labels that react with suitable substrates to produce breakdown products that may be chromogenic, fluorogenic, or luminescent; can be homogeneous or heterogeneous
Capture Assays (Sandwich Immunoassays)
Best suited for antigens with multiple determinants like antibodies, cytokines, proteins, tumor markers, and microorganisms
Fluorescent Immunoassay (FIA)
Involves qualitative observations using a fluorescent microscope, used for rapid identification of microorganisms, tumor-specific antigens, and transplantation antigens
Direct IFA
Antibody conjugated with a fluorescent tag added directly to unknown antigen fixed to a microscope slide for pathogen presence demonstration
Indirect IFA
Patient serum incubated with known antigen attached to a solid phase, then anti-human immunoglobulin with a fluorescent tag added to form a sandwich for antibody identification
Chemiluminescent Immunoassays
Follows antigen-antibody combination with light emission from a chemical reaction, can be used in heterogeneous and homogenous assays, sensitive with a large linear range of measurements
Rapid Immunoassays
Membrane-based, single-use, and disposable assays used for rapid strep, pregnancy, mono, etc., in home, clinic, or hospital settings
Immunoassay
Antigen or antibody coupled to membrane, producing visible color change
Colloidal immunoassay
Incorporates colloid for visible color change in reaction
Hybridization Methods
Techniques like Southern blot, Northern blot, array, and solution hybridization
Southern blot
Uses probe with known sequence to identify DNA sequence
Northern blot
Identifies RNA sequences
Array methods
Examines multiple genes simultaneously using specific probes on glass slides
Solution hybridization
Probe and single-stranded target in solution form RNA:DNA hybrids
FISH
molecular technique that uses fluorescent probes to detect specific DNA sequences or gene changes on chromosomes
Polymerase chain reaction
A laboratory technique to rapidly amplify millions of copies of specific DNA segments
PCR
In vitro replication process with oligonucleotide primers and synthetic nucleic acids
Denaturation
First step of PCR cycle: DNA separation at 94-96°C
Annealing
Second step of PCR cycle: Primer binding at 50-70°C
Extension
Third step of PCR cycle: DNA synthesis at 68-72°C
Transcription Based Amplification
Process focusing on making copies of RNA at a single temperature
Probe Amplification
Extending primers into multiple copies of probes with known target sequence
Signal Amplification
Binds large signal amounts to target sequences, detecting multiple genotypes