ISBB - IMMUNOLOGIC PROCEDURES Flashcards by Shynne Galang

Force of attration between a single Fab sute on an antibody molecule and a singke epitope on the corresponding antigen

Affinity

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Represents the overall streangth of antigen-antibidy binding

Avidity

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The ability of a particular antibody to combine with one antigen instead of another

Specificity

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It occurs if 2 different antigens share an identical or very similar epitope

Cross-reactivity

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Combination of antigen and antibody (sensitization)

Primary/Labeled Immunoassays

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Most sensitive and specific tests

Labeled Immunoassays

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Lattice formation and other demonstrable consequences of ag-ab reaction

Secondary/(precipitation/agglutination)

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Immunologically active in vivo reactions such as inflammation, chemotaxis, etc.

Tertiary/Skin test

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Preservation of serum if refrigeration is use

4°C for 72 hrs

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Preservation of serum if freezing

-20° C for longer period

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Destruction of complement at

56°C for 30 minutes

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Reinactivation of complement

4 hours after inactivation (56°C for 10 minutes)

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Chemical inactivation of Complement

Choline chloride

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It is the expression of relstive concentration

Dilution

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It is the volume of serum divided by total volume of solution

Simple dilution

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It is a dilution that involves 2 or more steps

Compound dilution

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It is a compound dilution in which the dilution factor is the sams in each step; used to obtain a titer

Serial dilution

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It is the reciprocal of the highest sefum dilution that gives a positive reaction with the antigen

Titer

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Both antigen and atibody are soluble

Precipation

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Zone of antibody excess

Prozone

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Zone of antigen excess

Postzone

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Zone of optimal antigen-antibody ratio

Zone of equivalence

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It measures the reduction in light intensity due to particles in a solution

Turbidimetry

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Measures the amount of light scattered at a particular angle from the incident beam as it passes through a suspension

Nephelometry

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Single diffusion, double dimension

Radial immunodiffusion

Single diffusion, single dimension

Ouidin Test

Double diffusion, single dimension

Oakley and Fulthorpe

Uses a plate containing agar with known antibody. Serum is placed on wells cut in the gel and the diameter of the precipitib ring formed is measured using either Fahey or Mancini

Radial immunodiffusion

RIA thag is Kinetic

Fahey method (FAK-ME)

RIA that is end point

Mancini (FAK-ME)

Double diffusion, double dimension

Ouchterlony technique

Smooth curve arc (ag in the sample is same as known antigen)

Identity

Spur formation (ag in spx has some similarities with known ag)

Partial Identity

Crossed lines (ag in the spx is different from known ag)

Non-identity

Physical attachment of antibody molecules to antigens on oarticles

Sensitization

Antiserum is incorporated into the agar and the uknown antigen is placed in the well and electrophoresed

Rocket electrophoresis

Antiserum is applied directly to the gel's surface rather than placed in a trough

Immunofixation electrophoresis

The height of rocket-shaped precipitation is ____ proportional to antigen concentration

Directly

Establishment of cross-links between sensitized particles amd antibodies resulting in aggregation

Lattice formation

Agglutination of particles artificially coated with antugens indicatiin the presence of antibody in the px sample

Indirect agglutination

Agglutination of particles with naturally occuring antigen on their surfaces

Direct agglutination

Agglutinatiin of antibody-coated staphylococcal cells by specific antigen

Coagglutination

Inhibition of agglutination of antigen coated particles by reagent antibody in the presence of homologous antigen in px sample

Agglutination inhibition (+ is no agglutination)

Agglutinatiin of particles artificially coated with antibody indicating the presence of antigeb in the patient's sample

Reverse passive agglutination

Involves migration of antigens and antibodies toward each other forming a precipitin line at equivalence

Counterimmunielectrophoresis

Sample used in complement fixation

Inactivated/Heated Serum

Complement best source

Guinea pig serum

Consist of sheep RBCs sensitized with hemolysin

Indicator system (best soruce: rabbit serum)

Positive result for complement fixation

No lysis

Positive result in NEUTRALIZATION

Absence of the corresponding reaction

Antigen-antibody reaction in which the biological effects of viruses and toxins are neutralized by homologous antibodies (neutralizing ab)

Neutralization

Neutralization of SLO inhibiting the lysis of indicator red cells

ASO titration

Positive for ASO titration

No hemolysis

Antibodies in oatient srfum neutralize viruses by preventing the formation of plaques (white patches of dead cells)

Plaque reductiin assay

Use of labeled antibody to detect unknown antigen (Labeled Immunoassay)

Direct

Amount of bound label is inversely proportional to the concentration of the analyte (Labeled Immunoassay)

Competitive

Amount of bound label is directly proportional to the analyte concentration (Labeled Immunoassay)

Non-competitive

Use of labeled antiglobulin to detect patient antibody (Labeled Immunoassay)

Indirect

Use of solid phase-bound antibody to detect the analyte of interest (Labeled Immunoassay)

Sandwich/Capture assay

Requires separation technique to remove unbound antigens or antibodies (Labeled Immunoassay)

Heterogenous

Does not require separation technique; consists only of a liquid phase (Labeled Immunoassay)

Homogenous

Detection method is for Scintillation counting

RIA

Colorimetry or Spectro following adhesion of chromogenic substances

EIA

FIA antigen detection in tissue or body fluids

Direct FIA

FIA antibody identification

Indirect FIA

A blocking test whose positive result is no fluorescence

Inhibition IFA

FTA-ABS/FANA

Inhibition IFA

Luminometry that uses ruthenium as label

ECLIA

Uses a compound that generate light when a voltage is applied to the electrode

ECLIA

Used to identify microbial antigens or antibodies, for oregnancy testing and for testing cardiac troponin

Rapid IA

Falsely low results when an extremely high antigen concentraion is present and the majority of binding sites are filled

Hook effect

Enzymes that recognize and bind to a specific nucleotide sequences in DNA

Restriction endonucleases

Differences in restriction patterns due to variations in nucleotide sequences

Restriction fragment length polymorphisms

Use of fluorescent labeled probe to detect target nucleic acid detected in intact cells

Fluorescent in situ hybridization

Heat stable DNA polymerase from Thermus aquaticus

Taq polymerase

Short segments of DNA designed to hybridize to template strands

Primers

Building blocks from which DNA strands are synthesized

Deoxyribosenucleoside triphosphates

Separation of template DNA to single strands

Denaturation

Binding of primers specific for each target strand sequence

Annealine

Addition of dNTPs to produce new strands

Elongation/Extension

Involves visualization of PCR products using gel electrophoresis

Conventional PCR

Uses fluorescent probes to monitorbthe accunulation of PCR product

Real-time/quantitative PCR

Used for cellular RNA or qualitative detection of RNA viruses

RT-PCR

Is performed to detect nultiple targets simultaeneously using more than one primer pair in a single reaction

Multiplex PCR

Uses 2 separate primer pairs in 2 sequential reaction

Nested PCR

Uses RNA probes complementary to the target DNA followed by capture and detrction of the resulting RNA; used to detect oncogenic variants of HPV

Hybrid capture assay

Uses a series of short single stranded DNA probes to capture the target nucleic acid and to bind to multiple reporter molecules

Branched DNA amplication

Used for qualitative and quantitative detection of HBV, HCV, and HIV-1

Branched DNA amplification

Measurement of light scattering and fluorescence as cells flow through a laser beam one at a time

Flow cytometry

For cell transportation via a carrier stream of isotonic saline that is hydrodynamically focused so that tge cells pass in a single file

Fluidics system

Plots a chosen parameter on the x axis versus the number of events in the y axis

Single parameter histogram

Plots 2 parameters against each other; each dot represents an individual cell or event

Dual parameter dot plot

Consists of cells that are positive for fluorescence on the y- axis and negative for fluorescence on the x axis

Quadrant 1

Consists of cells that are positive for fluorescence on both the x and y axis

Quadrant 2

Consists of cells that are negative for fluorescence on bith x and y axis

Quadrant 3

Consists of cells that are positive for fluorescence on the x axis and negative for fluorescnece on the y axis

Quadrant 4

Progenitor cells

CD 34

Plasma cells

CD 38

All hematopoietic cells

CD 45

ISBB - IMMUNOLOGIC PROCEDURES Flashcards

(100 cards)