antibody detection lecture presentation handout
DESCRIPTION
Blood Banking - Antibody detection and identification handout 2009TRANSCRIPT
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Antibody Detection dand
IdentificationIdentification
Michelle BrownMichelle Brown
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f dFrequency of unexpected antibodies in antibodies in hospitalized ppatients…2%
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Purpose of Antibody Detection and IdentificationIdentification…
To detect and identify antibodies that are clinically significantclinically significant
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What is a Clinically Significant y gAntibody?
An antibody that reacts at 37°C which can cause in vivo red cell which can cause in vivo red cell
destruction
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What if you Have a Clinically S f b d ?Significant Antibody?
1 Identify the antibody1. Identify the antibody2. Find antigen negative blood3. Crossmatch compatible
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What’s Clinically Significant?
Always Significant Sometimes SignificantAlways SignificantABORh
Sometimes SignificantLea
MN RhKellDuffy
MN P1
LutheranDuffyKiddSs
Lutheran
Rarely SignificantLeb
Xga
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How do we Know if the Patient h b d ?has an Antibody?
They have a positive antibody screenscreen.
ABO Discrepancy – AB with anti-MAA B A1 B cell
M + M -
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How do we know the Patient has an Antibody?
I tibl t hIncompatible crossmatchTransfusion reactionHDNPositive DATPositive auto control
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What do we need for Antibody Identification?
ABO d RhABO and RhAntibody screenResults of previous testingHistory of transfusion or pregnancyDiagnosisMedicationsMedications
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Reagent Red Cells - ABSCReagent Red Cells ABSC
G OGroup O
2-cell or 3-cell screens2 cell or 3 cell screens
Typed for all common antigens in j bl d tmajor blood group systems
Match lot # with antigen profile sheet Match lot # with antigen profile sheet (antigram)
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Required Testing ParametersRequired Testing Parameters
M lMay use serum or plasmaMethods that will demonstrate li i ll i ifi t tib diclinically significant antibodies
Must include 37°C incubation and AHGAHGPolyspecific or anti-IgGIgG presensitized cells (check cells)
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Antibody Panels
10 20 O t d ll10-20 group O reagent red cellsAntigram – indicates the phenotypes f th t d llof the reagent red cells
Pattern of reaction of patient serum i h ll i h d h f with cells is matched to the pattern of
antigen typing to determine the antibody specificityantibody specificity
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D C E c e M N S s K k Fya
Fyb
Jka
Jkb
IS 37 AGT
1 0 + 0 + + + 0 + + 0 + + 0 + 0 0 0 0
2 + + 0 0 + + + + 0 0 + + + + 0 0 0 0
3 + + + 0 + 0 + 0 + 0 + + + 0 + 0 0 +3 0 0 0 0 0 0 0 +
4 + 0 + + 0 0 + + 0 + 0 0 + + + 0 0 +
5 0 0 0 + + + + 0 + 0 + + 0 + + 0 0 05 0 0 0 + + + + 0 + 0 + + 0 + + 0 0 0
6 0 0 + + + + + + + 0 + 0 + 0 + 0 0 +
7 + 0 0 + + 0 + 0 0 + + 0 0 + 0 0 0 0
8 0 0 0 + + + 0 0 + 0 + + + 0 + 0 0 0Pt 0 0 0
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D C E c e M N S s K k Fya
Fyb
Jka
Jkb
IS 37 AGT
1 0 + 0 + + + 0 + + 0 + + 0 + 0 0 0 0
/ / / / / / / / // / / / /
2 + + 0 0 + + + + 0 0 + + + + 0 0 0 0
3 + + + 0 + 0 + 0 + 0 + + + 0 + 0 0 +3 0 0 0 0 0 0 0 +
4 + 0 + + 0 0 + + 0 + 0 0 + + + 0 0 +
5 0 0 0 + + + + 0 + 0 + + 0 + + 0 0 05 0 0 0 + + + + 0 + 0 + + 0 + + 0 0 0
6 0 0 + + + + + + + 0 + 0 + 0 + 0 0 +
7 + 0 0 + + 0 + 0 0 + + 0 0 + 0 0 0 0
8 0 0 0 + + + 0 0 + 0 + + + 0 + 0 0 0Pt 0 0 0
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D C E c e M N S s K k Fya
Fyb
Jka
Jkb
IS 37 AGT
1 0 + 0 + + + 0 + + 0 + + 0 + 0 0 0 0
/ / / / / / / / /
2 + + 0 0 + + + + 0 0 + + + + 0 0 0 0
3 + + + 0 + 0 + 0 + 0 + + + 0 + 0 0 +3 0 0 0 0 0 0 0 +
4 + 0 + + 0 0 + + 0 + 0 0 + + + 0 0 +
5 0 0 0 + + + + 0 + 0 + + 0 + + 0 0 05 0 0 0 + + + + 0 + 0 + + 0 + + 0 0 0
6 0 0 + + + + + + + 0 + 0 + 0 + 0 0 +
7 + 0 0 + + 0 + 0 0 + + 0 0 + 0 0 0 0
8 0 0 0 + + + 0 0 + 0 + + + 0 + 0 0 0Pt 0 0 0
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Evaluating Antibody Specificity
V i t th f ti itVarying strength of reactivityDosageM lti l tib diMultiple antibodiesVarying strength of antigen on red cells (P Le I Fy Jk D)(P, Le, I, Fy, Jk, D)
If difficult pattern, may phenotype the patients cells to see which the patients cells to see which antibodies they could make
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Panel Interpretation
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Some cells pos, some neg, auto neg
D C E c e M N S s K k Fya
Fyb
Jka
Jkb
IS 37 AGT
1 0 + 0 + + + 0 + + 0 + + 0 + 0 0 0 0
2 + + 0 0 + + + + 0 0 + + + + 0 0 0 0
3 + + + 0 + 0 + 0 + 0 + + + 0 + 0 0 +3 0 0 0 0 0 0 0 +
4 + 0 + + 0 0 + + 0 + 0 0 + + + 0 0 +
5 0 0 0 + + + + 0 + 0 + + 0 + + 0 0 05 0 0 0 + + + + 0 + 0 + + 0 + + 0 0 0
6 0 0 + + + + + + + 0 + 0 + 0 + 0 0 +
7 + 0 0 + + 0 + 0 0 + + 0 0 + 0 0 0 0
8 0 0 0 + + + 0 0 + 0 + + + 0 + 0 0 0Pt 0 0 0
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Interpretation of Typical Panel Results
Some cells pos some neg auto negSome cells pos, some neg, auto negSingle alloantibody
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Many or all cells pos, auto neg
D C E c e M N S s K k Fya
Fyb
Jka
Jkb
IS 37 AGT
1 0 + 0 + + + 0 + + 0 + + 0 + 0 0 0 +
2 + + 0 0 + + + + 0 0 + + + + 0 0 0 +
3 + + + 0 + 0 + 0 + 0 + + + 0 + 0 0 +3 0 0 0 0 0 0 0 +
4 + 0 + + 0 0 + + 0 + 0 0 + + + 0 0 +
5 0 0 0 + + + + 0 + 0 + + 0 + + 0 0 +5 0 0 0 + + + + 0 + 0 + + 0 + + 0 0 +
6 0 0 + + + + + + + 0 + 0 + 0 + 0 0 +
7 + 0 0 + + 0 + 0 0 + + 0 0 + 0 0 0 +
8 0 0 0 + + + 0 0 + 0 + + + 0 + 0 0 +Pt 0 0 0
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Interpretation of Typical Panel Results
Some cells pos some neg auto negSome cells pos, some neg, auto negSingle alloantibody
Many or all cells pos, auto negy p , gMultiple antibodiesAntibody to high incidence antigen
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Few cells pos, no evident patternD C E c e M N S s K k Fy
aFyb
Jka
Jkb
IS 37 AGT
1 0 + 0 + + + 0 + + 0 + + 0 + 0 + 0 0
2 + + 0 0 + + + + 0 0 + + + + 0 0 0 0
3 + + + 0 + 0 + 0 + 0 + + + 0 + 0 0 03 0 0 0 0 0 0 0 0
4 + 0 + + 0 0 + + 0 + 0 0 + + + 0 0 0
5 0 0 0 + + + + 0 + 0 + + 0 + + 0 0 05 0 0 0 + + + + 0 + 0 + + 0 + + 0 0 0
6 0 0 + + + + + + + 0 + 0 + 0 + 0 0 0
7 + 0 0 + + 0 + 0 0 + + 0 0 + 0 0 0 0
8 0 0 0 + + + 0 0 + 0 + + + 0 + + 0 0Pt 0 0 0
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Interpretation of Typical Panel Results
Some cells pos some neg auto negSome cells pos, some neg, auto negSingle alloantibody
Many or all cells pos, auto negy p , gMultiple antibodiesAntibody to high incidence antigen
Few cells pos no evident patternFew cells pos, no evident patternWeak antibodyAntibody showing dosageTechniques to enhance reactions?
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All cells pos, including auto controlD C E c e M N S s K k Fy
aFyb
Jka
Jkb
1 0 + 0 + + + 0 + + 0 + + 0 + 0 +
2 + + 0 0 + + + + 0 0 + + + + 0 +
3 + + + 0 + 0 + 0 + 0 + + + 0 + +3 0 0 0 0 0 +
4 + 0 + + 0 0 + + 0 + 0 0 + + + +
5 0 0 0 + + + + 0 + 0 + + 0 + + +5 0 0 0 + + + + 0 + 0 + + 0 + + +
6 0 0 + + + + + + + 0 + 0 + 0 + +
7 + 0 0 + + 0 + 0 0 + + 0 0 + 0 +
8 0 0 0 + + + 0 0 + 0 + + + 0 + +Pt +
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Interpretation of Typical Panel Results
Some cells pos some neg auto negSome cells pos, some neg, auto negSingle alloantibody
Many or all cells pos, auto negy p , gMultiple antibodiesAntibody to high incidence antigen
Few cells pos no evident patternFew cells pos, no evident patternWeak antibodyAntibody showing dosage
All cells pos, auto posCold autoantibodyWarm autoantibodyWarm autoantibodyRouleaux
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Selected Cells
If d t h th ll If you don not have the cells on your panel to rule out an antibody, you need to run additional cellsneed to run additional cells
If d h h ll If you do not have the cells on your panel to prove an antibody, you need to run additional cellsto run additional cells
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Methods of Testing for Antibody
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Low Ionic Strength Solution (LISS)
+ ++
++
+
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LISS
I iti itIncreases sensitivity
Don’t increase serum:cell ratio
Less sensitive method for detection of Kell antibodies
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PEG
P l th l l lPolyethylene glycolRemoves water, concentrates the
tib d t tib d t k antibody, promotes antibody uptake, and enhances strength of reactionD ’ ifDon’t centrifugeUse anti-IgG for AGT
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T t T bTest TubeGelSolid Phase Red Cell Adherence
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Gel Test: Pos test Neg testPos test Neg test
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Gel Test: Antibody Screeny
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Gel Test: ABO Typing
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Solid Phase Red Cell Adherence
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Solid Phase Red Cell Adherence