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1. IMMUNOHEMA PRELIM > TOPIC 1 ANTIGLOBULIN TEST PRELIM > Flashcards

TOPIC 1 ANTIGLOBULIN TEST PRELIM Flashcards

1
Q

aka COOMB’S TEST

A

ANTIGLOBULIN TEST

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2
Q

ANTIGLOBULIN TEST aka

A

COOMB’S TEST

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3
Q

Detects NON-AGGLUTINATING ANTIBODY by means of COUPLING with a SECOND ANTIBODY

A

COOMB’S TEST

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4
Q

Agglutination takes place because the _______ is able to BRIDGE the distance between cells that IgG alone cannot do

A

ANTIHUMAN GLOBULIN

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5
Q

Demonstrate IN VIVO COATING of red cells with ANTIBODIES or COMPLEMENT

A

DIRECT ANTIGLOBULIN TEST

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6
Q

Detects PRESENCE of “UNEXPECTED” ANTIBODIES in the patient’s SERUM that COAT ANTIGENS in TYPE-O red cells in VITRO

A

INDIRECT ANTIGLOBULIN TEST

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7
Q

DIRECTLY AGGLUTINATES RBCS suspended in SALINE

A

IgM

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8
Q

NONAGGLUTINATING or INCOMPLETE ANTIBODIES because their MONOMERIC STRUCTURE

A

IgG

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9
Q

can synthesize/coat RBC without visible agglutination

A

IgG

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10
Q

MAJOR TYPES OF BLOOD GROUP ANTIBODIES

A

IgM
IgG

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11
Q

ANTIHUMAN GLOBULIN REAGENTS

A

Polyspecific AHG reagent
Monospecific AHG reagent

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12
Q

AHG reagent that
Contain antibody to HUMAN IgG and to the C3D COMPONENT of the HUMAN COMPLEMENT

A

POLYSPECIFIC AHG Reagents

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13
Q

AHG reagent that FACILITATE AGGLUTINATION

A

POLYSPECIFIC AHG Reagents

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14
Q

AHG reagent that is more specific

A

Monospecific AHG reagent

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15
Q

Contain ONLY ONE ANTIBODY SPECIFICITY

A

MONOSPECIFIC AHG Reagents

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16
Q

Monospecific AHG reagent contain antibodies either??

A

Anti-IgG or Anti-C3B/Anti-C3D

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17
Q

breaks the bonds between the antibody and antigen

A

Elution

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18
Q

If positive in monospecific and polyspecific =

A

Elution

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19
Q

If positive in polyspecific = confirm by? [polyspecific (+), monospecific (-)]

A

MonoSpecific
Confirm by adding O CHECK CELLS (Coombs cell)

*If (+) agglutination: VALID
(Indicates that AHG is added and AHG reagent was not NEUTRALIZED)

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20
Q

AHG reagent bind to the FC region of the antibody

A

C3D- degradation

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21
Q

TYPES OF AHG PREPARATION

A
  • CLASSIC METHOD
  • POLYCLONAL ANTIBODIES
  • MONOCLONAL ANTIBODIES
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22
Q

METHOD
* Injecting human serum or purified globulin into laboratory animals (RABBITS)
* Human globulin behaves as FOREIGN ANTIGEN
* The RABBIT’S IMMUNE RESPONSE is TRIGGERED
* An ANTIBODY to HUMAN GLOBULIN is PRODUCED

A

CLASSIC METHOD

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23
Q

Method
Mixture of antibodies from DIFFERENT PLASMA CELL CLONES which can recognize different ANTIGEN DETERMINANTS or the SAME PORTION of the ANTIGEN but with DIFFERENT AFFINITIES

A

POLYCLONAL ANTIBODIES

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24
Q

Method
* Derived from ONE CLONE of PLASMA CELLS and RECOGNIZE a SINGLE EPITOPE

A

MONOCLONAL ANTIBODIES

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25
3 Antibody molecules and Complement components
* Alpha * Beta * Gamma
26
Injecting an animal with HUMAN GLOBULIN stimulates the animal to _____ to the _______
REPRODUCE ANTIBODY FOREGIN PROTEIN
27
employ a variety of AHG reagents REACTIVE with various HUMAN GLOBULINS
Serologic Tests
28
aka DIRECT COOMB’S TEST
DIRECT ANTIGLOBULIN TEST
29
Detects IN VIVO SENSITIZATION of RBCs with IgG or COMPLEMENT COMPONENTS
DIRECT ANTIGLOBULIN TEST
30
Clinical conditions that can result in IN VIVO COATING of RBCs with ANTIBODY COMPLEMENT are:
HDN HTR AIHA DIHA
31
fetal cells are coated with maternal antibodies in vivo
HDN
32
patients RBC coated with antibodies in vivo
HTR (Hemolytic Transfusion Reaction)
33
o 4 mechanisms that results to DAT positive
Drug Adsorption Membrane Modification Formation of Immune Complex Autoantibody Formation
34
Aka INDIRECT COOMB’S TEST
INDIRECT ANTIGLOBULIN TEST
35
* DETECTS IN VITRO SENSITIZATION OF RBCS
INDIRECT ANTIGLOBULIN TEST
36
USED IN THE FOLLOWING SITUATIONS: * DETECTION OF INCOMPLETE ANTIBODIES TO POTENTIAL DONOR RBCs OR TO SCREENING CELLS IN SERUM. * DETERMINATION OF RBC PHENOTYPE USING KNOWN ANTISERA. * TITRATION OF INCOMPLETE ANTIBODIES
INDIRECT ANTIGLOBULIN TEST
37
DAT Specimen: Test: Tube:
RBC Antigen Phenotyping Pink stopper/ or EDTA
38
IAT Specimen: Test: Tube:
Serum (Unbound) Crossmatching Antibody detection Antibody identification Red Stopper
39
* Free-floating in plasma
INDIRECT ANTIGLOBULIN TEST
40
* Antibodies that are bound, for autoimmune diagnosis
DIRECT ANTIGLOBULIN TEST
41
IAT AS A PROCESS
Patients serum + RBC with know antigen After combining, wash If added, allow antibodies to attach the RBC surface Add AHG reagent that bind to the FC region of antibodies to produce agglutination
42
Why do we wash RBC?
To remove unwanted proteins, antibodies, and electrolytes that can cause adverse reactions
43
FACTORS AFFECTING ANTIGLOBULIN TEST
Ratio of serum to cells Reaction Medium Temperature Incubation time Washing of RBC Addition of AHG Centrifugation for reading
44
Increasing the ratio of serum to cells INCREASES the ____ of the test system
SENSITIVITY
45
Target when doing AHG test:
40:1 2 drops of serum + 1 drop 5% volume of solute/volume of solution cell suspension
46
allow AB-COATED CELLS to come into CLOSER CONTACT with each other
Albumin
47
ENHANCE AB UPTAKE and allow INCUBATION TIME to be DECREASED
Low Ionic Strength Solution
48
Low Ionic Strength Solution Incubation time
10-15 minutes
49
Reaction medium that causes aggreation to RBC
Polyethylene glycol
50
INCREASE ANTIBODY UPTAKE by REDUCING WATER MOLECULES surrounding RBC
Polyethylene glycol
51
what are the 3 reaction mediums
Albumin Low Ionic Strength Solution Polyethylene Glycol
52
OPTIMAL temperature for complement activation and rate of reaction
37C
53
Why is 37C the optimal temperature for AHG testing
The rate of reaction depends on this temperature
54
INCUBATION TIME for AHG testing
30-120 minutes
55
If Incubation time is extended upto 40 minutes or more? what happens?
Decrease Test Sensitivity
56
inadequate washing causes?
False Negative RXN
57
MOST IMPORTANT PHASE in testing
Washing phase
58
controlled by CHECK CELLS or GROUP-O CELLS sensitized with IgG, done immediately
WASH PHASE
59
should be DISCARDED AFTER the FINAL WASH
Saline
60
WASHING OF RBCs
Wash for a minimum of 3x at 37C
61
nadequate washing – FALSE-NEG RXN because of the ______ by ______
NEUTRALIZAITON of AHG reagent RESIDUAL UNBOUND SERUM GLOBULINS)
62
AHG reagent should be added immediately after washing to minimize?
Antibody eluting from the cell and NEUTRALIZE AHG reagent
63
What does higher RCF do?
yield more sensitive results
64
INADEQUATE RESUSPENSION result to?
Weak False-Positive
65
Too vigorous resuspension results to?
False Negative
66
Give 3 Sources of error for False Positive Results
Over incubation Over centrifugation Dirty Glassware * Contaminated reagent * Direct agglutination by agglutinins * Improper use of enhancement RGTS * Saline stored in GLASS or METAL (PLASTIC) * Presence of FIBRIN (CLOT)
67
Give 3 Sources of error for False Negative Results
Delayed Washing Expired Reagent Improper Centrifugation * Improper washing (VIGOROUS SHAKING) * Failure to add AHG reagent * Improper centrifugation * Serum cell ratio TOO LOW (40:1) / (133:1)
68
What are the MODIFIED AND AUTOMATED ANTIGLOBULIN TEST TECHNIQUES
Low Ionic Polybrene Technique Solid-Phase Technology Enzyme-Linked Antiglobulin Test Gel Test
69
MODIFIED AND AUTOMATED ANTIGLOBULIN TEST TECHNIQUES Relies on LOW IONIC CONDITITONS to RAPIDLY SENSITIZE CELLS with ANTIBODY
LOW IONIC POLYBRENE TECHNIQUE
70
MODIFIED AND AUTOMATED ANTIGLOBULIN TEST TECHNIQUES is added to allow the SENSITIZED CELLS to approach each other and PERMIT CROSS-LINKING by the attached antibody
Polybrene
71
MODIFIED AND AUTOMATED ANTIGLOBULIN TEST TECHNIQUES * POLYBRENE is added to allow the SENSITIZED CELLS to approach each other and PERMIT ____ by the attached antibody
CROSS-LINKING
72
What solution is added to reverse Rouleaux
High Ionic Strength Solution
73
MODIFIED AND AUTOMATED ANTIGLOBULIN TEST TECHNIQUES * TEST CAN BE CARRIED THROUGH TO AN AHG TECHNIQUE IF REQUIRED * MONOSPECIFIC ANTI-IgG reagent MUST BE USED
LOW IONIC POLYBRENE TECHNIQUE
74
Types of Solid-Phase Technology
Direct Indirect
75
In Direct SOLID-PHASE TECHNOLOGY Antibody is attached to a?
Microplate Well
75
In Indirect SOLID-PHASE TECHNOLOGY * Known RBCs are BOUND to a WELL that has been treated with ???
GLUTARALDEHYDE
76
In Direct SOLID-PHASE TECHNOLOGY If antibody is SPECIFIC FOR ANTIGEN on RBCs, the BOTTOM OF THE WELL will be COVERED with ?
Suspension
77
In Direct SOLID-PHASE TECHNOLOGY * If NO SUCH SPECIFICITY OCCURS, RBCs will ____ to the BOTTOM OF THE WELL
SETTLE
78
In Indirect SOLID-PHASE TECHNOLOGY * If antibody in SERUM IS SPECIFIC FOR ANTIGEN on FIXED RBCs, a _____ OCCURS
POSITIVE REACTION
78
In Indirect SOLID-PHASE TECHNOLOGY what is added to RBC-COATED WELLS
TEST SERUM
79
MODIFIED AND AUTOMATED ANTIGLOBULIN TEST TECHNIQUES * An RBC suspension is added to MICROTITER WELL and washed with SALINE then AHG is added
ENZYME-LINKED ANTIGLOBULIN TEST (ELAT)
80
In ELAT * The enzyme-labeled AHG will BIND to ______
IgG-SENSITIZED RBCs
81
MODIFIED AND AUTOMATED ANTIGLOBULIN TEST TECHNIQUES * An RBC suspension is added to MICROTITER WELL and washed with SALINE then AHG is added * The enzyme-labeled AHG will BIND to IgG-SENSITIZED RBCs * Excess antibody is REMOVED and ENZYME SUBSTRATE is added * The amount of COLOR PRODUCES is MEASURE SPECTROPHOTOMETRICALLY and is PROPORTIONAL to the ANTIBODY PRESENT
ENZYME-LINKED ANTIGLOBULIN TEST (ELAT)
82
TYPES OF GEL TESTS
* Neutral GEL Test * Specific GEL Test * GEL Low Ionic Antiglobulin Test
83
MODIFIED AND AUTOMATED ANTIGLOBULIN TEST TECHNIQUES Process that DETECTS RBC ANTIGEN-ANTIBODY REACTIONS by means of using a CHAMBER filled with POLYACRYLAMIDE GEL
GEL TEST
84
In Gel Test are Process that DETECTS RBC ANTIGEN-ANTIBODY REACTIONS by means of using a CHAMBER filled with??
POLYACRYLAMIDE GEL
85
In Gel Test The Gel acts as a??
Trap;Free Unagglutinated RBCs form pellets in the Bottom of the Tube
86
In Gel Test * AGGLUTINATED RBCs are TRAPPED in the TUBE for ____?
HOURS
87
In Gel Test NEGATIVE REACTIONS appear ?
Pellets in the bottom of the microtube
88
In Gel Test POSITIVE REACTION is?
Fixed in the GEL

1. IMMUNOHEMA PRELIM (2 decks)

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