Week 2 Flashcards

(40 cards)

1
Q

What occurs in HDFN

A

When mom is negative and baby is positive, the first pregnancy has mom produce Anti-D from the being exposed to D positive cells
-Maternal anti D are able to cross placenta
-the fetal cells in the next pregnancies are destroyed by AntiD if the baby is Rh pos

-mom is protected with Rh neg against the anti D after delivery

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2
Q

What is the difference between the fisher race theory and Wiener Theory

what is the current theory

A

Fisher-Race Theory:
A gene complex is inherited that codes for 3 closely linked set of alleles

Wiener Theory:
1 gene is responsible for the expression of all
Rh Blood Group systems

current :Rh Blood Group
System antigens are determined By 2 genes. Allele D or RHCE

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3
Q

What are Rh Genetics

A

-genes codominant alleles
-RHD= D Ag as heterozygous or homozygous Dd or DD

RHCE genes have 4 alleles at am locus
1) RHCE
2) RHCe
3) RHcE
4) RHce

5 Ag can be found in most ppl

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3
Q

Which AG is most immunogenic

A

D Ag after A and B antigen
-Anti D is seen only if the pt with D Ag is exposed to D pos cells
-Exposure from pregnancy or transfusions

Rh pos/neg is whether there is D AG on RBC

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3
Q

Rh Antibodies- how are they different from ABO system

A

1) Red cell immune: DO NOT produce the antibodies unless exposed to red blood cells through transfusion or pregnancy
2) IgG
3) Poor Complement Binding

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3
Q

What is the structure of Rh Ag

A
  • protein structure-amino acids
    -Elicits an IgG immunogenic response
    -Exposure through transfusion or pregnancy-Red Cell Immune

 Not expressed on tissues
 Well-developed at birth
 Direct gene product (not enzyme)
 RHD/RHCE Genes codes directly for antigen
 Can easily cause Hemolytic Disease of the Fetus and Newborn (HDFN)

-protein that is made up of over 400 AA and crosses RBC membrane 12 times
-Exposed loops are the AG
-Product of RHD gene and RHCE gene

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3
Q

Rh Phenotype vs. Genotype

A

D+C+E-c+e+
Antigens Present, Serologically Detectable

DCe/Dce, Dce/dCe, DCe/dce
Genotype- Possible Genes Inherited

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3
Q

most common antibody in RH neg pt vs Rh pos

A

Anti D in Rh neg

Anti E in Rh pos

Anti e is autoAB hard to find since 98% of pop has e AG

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3
Q

Fisher-Race Nomenclature

A
  • 3 separate genes and their alleles D,C,E, c and e
    -linked on the same chromosome and inherited as a gene complex

AG that are possible are D,C,E, c and e
-d is amorphic or deleted gene
-If D antigen is present, the genotype can be either DD or Dd

DCE, DCe, DcE, Dce, dCE, dCe, dcE, dce

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4
Q

Wiener Nomenclature

A

One gene responsible for the expression of 8 alleles which causes many Rh Ag to be present
-terminology is still used even if the theory was incorrect

Ro, R1, R2, Rz, r, r’, r”, ry

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5
Q

Types of Rh Reagents
Monoclonal Anti-D

A

Monoclonal Anti-D and Polyclonal Anti-D blend – Must be mixed together for Weak D detection

Monoclonal Anti-D – Low protein diluent (6% bovine albumin) containing IgM

at michener we use Anti-D1 and Anti D2

Polyclonal Anti-D Blend – IgG and IgM

-Low protein diluent doesnt promote false positive agglu that you find with high protein D typing reagents

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6
Q

Types of Rh Reagents
Rh Control

A

-protein media can cause false pos
-Rh Control – Low protein diluent (6% bovine albumin) but NO anti-D
Purpose – Detects false positive results. To show reaction is due to anti-D NOT diluent
Should be NEGATIVE to report ABO and Rh typing

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7
Q

When would you have a positive Rh control

A

Positive DAT - AB on the cells (used modified anti D like monoclonal, saline AntiD)

Rouleaux/cold agglutinins - use 3x washed RBCS

Bacterial contamination - repeat

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8
Q

What are three types of weak D

Genetic Weak D

A

Genetic Weak D

D occurs when there are low AG

 Gene codes for less D antigens
 Usually needs IAT Weak D testing
 Does not usually produce Anti-D

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9
Q

What are three types of weak D

C Trans (positional)

A

 Position effect: C antigen inherited in
trans position to D antigen
 Weaker expression of D antigen
 Monoclonal Anti-D can detect
 Does not usually produce Anti-D

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9
Q

What are three types of weak D

Partial D or Mosaic D

A

Missing part of the Rh antigen
 Mutated gene: change in aa, therefore change in protein
 Several variations
 Detected by monoclonal Anti-D or may need Weak D testing (IAT)
 different results with different antisera (polyclonal vs monoclonal)
 Individuals can make Anti-D to the part of the antigen missing

if they are either they need to be treated as Rh neg because you dont want them to produce AB /Anti D

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10
Q

How to detect Weak D?

A

Antisera
-Immediate spin method may have a Weak Positive results- still call Rh Positive
-Immediate spin method may have a Discrepancy in testing
-If automation is used- discrepancy in D1/D2 testing

11
Q

Weak D Policies

A

initial Rh typing on everyone
 If the IS Anti-D is negative :
1) Stop if adult patient, result as Rh Negative

2) Perform Weak D testing on ALL Donors (at CBS) if its D1 and D2 neg then you have to know its actually neg

3) Perform Weak D testing on Rh Negative babies of Rh Negative Moms
IF Weak D testing is POSITIVE, then donors and babies are called Rh Positive.

12
Q

Weak D IAT test

A

To detect these weaker expressions of
the D antigen, we use the IAT method
- do normal D1,d2 if there is less AG then you can have sensitization without agglutination
-incubate Anti D and wash to remove extra antisera, and add ahg
-the AHG will show agglutination if there is sensitization

NO checking under the Microscope
Add CCC to all negative reactions
minimum reaction should be 1+

13
Q

Weak D IAT False positive

A

Weak D testing can be false positive due to in vivo coating of Red cells with IgG antibody
-Rh control is positive
-AHG reacts with the IgG on the RBC membrane even if there is not Anti D attached
-invalid weak D testing

Auto Antibody, Mom’s Antibody on Fetal
cells, and Patient Antibody on donor cells.

14
Q

Molecular Testing for Weak D
policy for testing

A
  • prenatal pts with weak serological RHD results were RHD genotyping can modify their blood product or RhIG needs
    -pt that need chronic transfusions or have complex needs where genotyping can change blood requirements
    -pt who appear serologically D positive but need to be transfused with AntD

-mosiac D is most important because the pt can produce Anti D

15
Q

how to select Rh for transfusion

A
  • Rh AB are red cell immune
    -Rh neg ppl dont automatically make Anti D
    -Rh neg pts can be given Rh pos RBC once safely the AB is produced quickly - antigenic
    -Anti D is an IgG and doesnt bind complement so hemolysis is extravascular (5-10 days)
    -C, E, c, or e antigens not considered unless the pt has corresponding AB
16
Q

What are ABO ABs

A

non red blood cell stimulated
-most clinially significant
-ppl have ABO AB for the AG they lack
-IgM
-activate complement, only one IgM is needed to initiate complement pathway
* Immediate Cell Lysis
* Intravascular Hemolysis
-can cause severe transfusion reactions

17
Q

Landsteiner’s Rules

A

1) A person does not have the antibody to their own antigen.
2) Each person has antibodies to the antigen, they lack (only in the ABO system)

18
ABO Genes where are they found
- on chromosome 9 -one allele from each patient -A and B are co dominant -o gene has no A or B AG, and its expressed until the person is OO
19
What are punnett squares
used to determine inheritance possibilities
20
ABO Antigens are found where
* RBC Membranes * Endothelial Cells * Platelets * Lymphocytes * Tissue * Soluble form found in plasma and other body secretions in a % of populations who have inherited the Secretor gene * Several genes and their enzymes are involved in the antigen production.
21
A and B genes code for?
enzymes = transferases -transfer sugar to a basic precursor substance on the RBC membrane -ABO antigens are carbohydrates attached through the action of a transferase enzyme Type 1 Precursor Substance – Beta1-3 linkage (ex. Secretory status) Type 2 Precursor Substance – Beta1-4 linkage (ex. RBCs)
22
The A & B Antigen Structure
-terminal sugars at the end of the long sugar chains (oligosaccharides) attached to lipids on the RBC membrane -last sugars added to the chain -O doesnt have the A/B ag (has the most amount of the last terminal sugar called H antigen) -Ags in different blood groups have the same precursor
23
Antigen Development
-A, B, and H antigens are controlled by the action of transferases. -*A, B, and H genes each produce a different transferase, adding a specific sugar to the chain. H Gene – Codes for Fucosyltransferase (FUT 1); Adds L-Fucose to Precursor Substance A Gene – Codes for N-Acetyl-galactosaminyltransferase; Adds N-Acetylgalactosamine to A Antigen B Gene – Codes for D-Galactosyltransferase; Adds D-Galactose to A Antigen NOT all H antigens are converted in A, B, or AB individuals * IF both A and B genes are present, some H chains are converted to A antigen, while others are converted to B antigen.
24
OO Genotype what enzyme or Ag is present
- neither enzyme is produced -doesnt convert any precursors to A or B AG -all have “unconverted” H antigen while A1B have the least H AG -O gene is an amorph (does not express a detectable product). -Blood Group O is recessive; the trait is expressed when inherited by both parents. O has most amount of H AG and AB has least
25
Rare and subtypes
26
Landsteiner’s Rules for ABO Antibodies
In the ABO blood group system, individuals possess ABO antibodies to the ABO blood group antigens they lack
27
What is the current hypotheses for ABO ab
--no known exposure and thought to be naturally occurring or stimulated by other substances -bacteria or pollen could have structures similar to A/B ag -environmental exposure would allow ppl to respond immunogenically and produce the AB -exposure could begin at birth -Non red cell immune -newborns cant produce own ABO Ab until they are 3-6 months - maternal AB -Do not perform RA/RB (Reverse Grouping) if less than 4 months of age.
28
What AB do O group ppl make
- Anti- A, B (IgG) -reacts with A or B Ag -cant be separated into Anti A or B
29
in routine BB how do we determine ABO GROUP
-Reverse cells RA and RB have the known antigen and are used to test with the patient plasma to determine the unknown antibodies (A1 cells and B cells).
30
4 Basic categories of Reagents and basic method of testing
1) Red Blood Cells with known antigens 2) Antisera with known antibodies 3) Potentiators to enhance antibody reactions 4) Antihuman Globulin reagents (Polyclonal and Monoclonal) 2 basic methods of testing immediate spin - IgM Antiglobulin test - IgG
31
Commercial Antibody Reagents * Polyclonal antibodies vs monoclonal
POLY -made from different clones of B cells with secrete AB with diff specificities -recognize diff epitopes -mix of both IgG or IgM AHG MONO- made from single clone of transformed B cells that secrete AB of SAME specificity -one immunoglobulin class IgG or IgM Uses Hybridoma technology * Recognizes a single epitope * E.g., Anti-C, Anti-A, and Anti-IgG
32
ABO Red Cell Testing (ABO Forward Testing)
Anti-A, Anti-B, and Anti-A,B are reagents used to determine the ABO blood type, antisera toward specific AG
33
ABO Typing forward vs reverse
forward- test pt cell for AG Reverse - test pt serum for AB