ABO Blood Grouping & Rh(D)

groups

Introduction
David Ntiamoah Ofosu; MSc,
CEMBA, PGCe, BSc

4/9/2015 DNO 1
The ABO System
Discovered in 1901 by Dr. Karl
Landsteiner
4 Main Phenotypes (A, B, AB, O)
ABO gene located on long arm of
chromosome 9

4/9/2015 DNO 2
ABO Basics
Blood group antigens are actually sugars
attached to the red blood cell.
Antigens are “built” onto the red cell.
Individuals inherit a gene which codes for
specific sugar(s) to be added to the red cell.
The type of sugar added determines the blood
group.

4/9/2015 DNO 3
 The ABO Antigens
Added to Proteins or Lipids in Red Cells
Substrate Molecule is H (fucose)
A antigen is N-acetyl-galactosamine (GalNAc)
B antigen is Galactose (Gal)
A and B genes code for transferase
enzymes

4/9/2015 DNO 4
This diagram illustrates the terminal sugar for each
blood group.

4/9/2015 DNO 5
ABO Antibodies
A and B substances very common
Antibodies produced to “non-self”
Produced after first few months of life
A & B people have mainly IgM
O people have IgG
May fade in old age

4/9/2015 DNO 6
Landsteiner’s Rule
Individual’s will form immune antibodies to ABO
blood group antigens they do not possess.
Substances are present in nature which are so
similar to blood group antigens which result in the
constant production of antibodies to blood group
antigens they do not possess.
Critical for understanding compatibility between ABO
blood groups.

4/9/2015 DNO 7
Antigens & Antibodies
Antigens on RBCs
Blood Group Antibodies in Serum Genotypes

A A Anti-B AA or AO

B B Anti-A BB or BO

AB A and B Neither AB

O Neither Anti-A and anti-B OO

4/9/2015 DNO 8
Inheritance of ABO Groups

Allele from Allele from Genotype of Blood types of

the mother the father offspring offspring

A A AA A

A B AB AB

A O AO A

B A AB AB

B B BB B

B O BO B

O O OO O

4/9/2015 DNO 9
ABO Typing
Cell Group Reverse Group
• Test Washed Cells • Test plasma/serum
With: with:
• Monoclonal Anti-A • Known A1 cells
• Monoclonal Anti-B • Known B cells
• Inert control • Known O cells
• ? Known A2 cells
Agglutination is a
positive result Reactions may be
weaker than cell
group
4/9/2015 DNO 10
Distribution of ABO Groups

Population O A B AB
Aborigines 61 39 0 0
Basques 51 44 4 1
Blackfoot (N. Am. Indian) 17 82 0 1
Bororo 100 0 0 0
Chinese-Canton 46 23 25 6
Chinese-Peking 29 27 32 13
English 47 42 8 3
Hawaiians 37 61 2 1
Irish 52 35 10 3
Mayas 98 1 1 1
Navajo (N. Am. Indian) 73 27 0 0
Peru (Indians) 100 0 0 0
United Kingdom (GB) 47 42 8 3
USA (blacks) 49 27 20 4
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USA (whites) 45 40 11 4
Distribution of the A allele

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Distribution of the B Allele

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Distribution of the O Allele

4/9/2015 DNO 14
ABO Type Frequencies In U.S.
ABO Type Per Cent

O 45%

A 40%

B 11%

AB 4%

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Significance of ABO Group
ABO mismatched transfusions:
• Rare
• May be life threatening
• Can be caused by technical or clerical error
• Intravascular haemolysis
• More severe in group O patients

4/9/2015 DNO 16
Antibody clinical significance
Immunizations are frequently done to protect us
from disease.
• Receive Hepatitis B immunization.
• Actual bits of hepatitis virus injected.
• Body recognizes as foreign and produces an immune
antibody.
• Subsequent exposure to real Hepatitis B virus will result in
destruction of the virus by immune antibodies.
ABO antibodies are immune and will result in
destroying incompatible cells which may result in the
death of the recipient.

4/9/2015 DNO 17
Genetics
Two genes inherited, one from each parent.
Individual who is A or B may be homozygous or
heterozygous for the antigen.
Heterozygous: AO or BO
Homozygous: AA or BB
Phenotype is the actual expression of the genotype,
ie, group A
Genotype are the actual inherited genes which can
only be determined by family studies, ie, AO.

4/9/2015 DNO 18
Inheritance
Blood group antigens are “codominant”, if
the gene is inherited, it will be expressed.
Some aberrant genotypes do occur but due
to the rarity will not be discussed.
Understanding of basic inheritance
important.

4/9/2015 DNO 19
Example of Determining
Genotype
Mom’s phenotype is group A, genotype AO
Dad’s phenotype is group B, genotype BO

B O
A AB 25% AO 25% (Group A)
O BO 25% (Group B) OO 25% (Group O)

4/9/2015 DNO 20
Other Examples
Mom Dad Offspring Blood
Group
AA BB 100% AB

BO OO 50% each of B or
O
OO OO 100% O

OO AO 50% each of A or
O
4/9/2015 DNO 21
Group O
Approximately 45% of the
population is group O.
No A or B antigens present,
think of as “0” antigens present.
These individuals form potent anti-
A and anti-B antibodies which
circulate in the blood plasma at all
times.

4/9/2015 DNO 22
Group A
Approximately 40% of the
population is group A.
No B antigens present.
These individuals form
potent anti-B antibodies
which circulate in the blood
plasma at all times.

4/9/2015 DNO 23
Group B
Approximately 11% of the
population is group B.
No A antigens present.
These individuals form potent
anti-A antibodies which circulate
in the blood plasma at all times.

4/9/2015 DNO 24
Group AB
Approximately 4% of the
population is group AB.
Both A and B antigens
present.
These individuals possess no
ABO antibodies.
NOTE: This slide is in error
as it only illustrates presence
of one antigen not 2.

4/9/2015 DNO 25
Hemolysis
If an individual is transfused with an incompatible
blood group destruction of the red blood cells will
occur.
This may result in the death of the recipient.

4/9/2015 DNO 26
Summary
Blood Group Antigens on cell Antibodies in Transfuse with
plasma group

A A Anti-B A or O

B B Anti-A B or O

AB A and B none AB, A, B or O

O None Anti-A & B O

4/9/2015 DNO 27
Universal Donor and Recipient
Universal Donor Universal Recipient
Group O Group AB
• Carries no A or B • Patient has no anti-A
antigens or anti-B present
• Packed and • Cannot lyse any
processed units transfused cells
have little antibody • Beware: other
• antibodies may be
present
Using the patient’s own group ASAP will conserve resources

4/9/2015 DNO 28
Rh (D) Antigen
Of next importance is the Rh type.
• Term “Rh” is a misnomer.
• Rh is a blood group system with many antigens,
one of which is D.
• Re-education of public is difficult.
Rh refers to the presence or absence of the
D antigen on the red blood cell.

4/9/2015 DNO 29
The Rh(D) Antigen
RH is the most complex system, with
over 45 antigens
Discovered in 1940 after work on
Rhesus monkeys
Subsequently discovered to be
unrelated to monkeys
RH gene located on short arm of
chromosome 1
4/9/2015 DNO 30
Simple Genetics of Rh(D)
86% of caucasians are Rh(D) pos
The antithetical antigen d has not been
found
The d gene is recessive:
• Dd, dD, DD, persons are Rh(D) pos
• Only dd persons are Rh(D) neg

4/9/2015 DNO 31
Rh (D) Antigen (continued)
Unlike the ABO blood group system, individuals who
lack the D antigen do not naturally make it.
Production of antibody to D requires exposure to the
antigen.
The D antigen is very immunogenic, ie, individuals
exposed to it will very likely make an antibody to it.
For this reason all individuals are typed for D, if
negative must receive Rh (D) negative blood.

4/9/2015 DNO 32
Rh (D) Antigen (continued)
The most important patient population to
consider is females of child-bearing age.
If immunized to Rh (D) antigen the antibody can
cross the placenta and destroy Rh (D) positive
fetal cells resulting in death.
This is why Rh negative women are given
Rhogam after birth of Rh positive baby.

4/9/2015 DNO 33
Hemolytic Disease of the Neborn
– How it Occurs
A child is Rh pos
B during pregnancy fetal Rh pos rbc’s escape into maternal
circulation
C Mother produces antibodies to Rh (D) antigen
D Second pregnancy with Rh (D) pos child results in
destruction of fetal D pos rbcs

4/9/2015 DNO 34
Blood Typing
There are 2 components to blood typing:
• Test unknown cells with known antibodies
• Test unknown serum/plasma with known rbc’s
The patterns are compared and the blood
group is determined.

4/9/2015 DNO 35
Slide Blood Typing
Very rudimentary method for determining blood
groups.
CANNOT be used for transfusion purposes as false
positives and negatives do occur.
A “false positive” is when agglutination occurs not
because the antigen is present, but cells may already
be clumpled.
A “false negative” is one in which the cells are not
clumped because there are too many cells or not
enough reagent.
4/9/2015 DNO 36
Slide Blood Typing - continued
The slide is divided into halves.
On one side a drop of anti-A is added, this will attach to
and cause clumping of rbcs possessing the A antigen.
On the other side a drop of anti-B is added which will
cause clumping of rbcs with the B antigen.
A drop of rbcs is added to each side and mixed well with
the reagent.
The slide is tilted back and forth for one minute and
observed for agglutination (clumping) of the rbcs

4/9/2015 DNO 37
Interpretation of Slide Typing Testing with
Anti-A Anti-Serum
If an rbc contains the A antigen
the red blood cells will be
agglutinated by anti-A, a positive
reaction.
If an rbc does not have the A
antigen there will be no clumping,
a negative reaction.

4/9/2015 DNO 38
Interpretation of Slide Typing Testing with
Anti-B Anti-Serum
If an rbc contains the B antigen the
red blood cells will be agglutinated
by anti-B, a positive reaction.
If an rbc does not have the B antigen
there will be no clumping by anti-B, a
negative reaction.

4/9/2015 DNO 39
Slide Blood Typing Group A
An unknown rbc suspension is added to known anti- sera.
The left hand of the slide contains anti-A which reacts with
the unknown cell.
The right hand side contains anti-B which does not
react with the cell.

4/9/2015 DNO 40
Slide Blood Typing Group B
An unknown rbc suspension is added to known anti- sera.
The left hand of the slide contains anti-A does not react with
the unknown cell.
The right hand side contains anti-B which reacts with
the cell.

4/9/2015 DNO 41
Slide Blood Typing Group O
The left hand of the slide contains anti-A does not
react with the unknown cell.
The right hand side contains anti-B does not react
with the unknown cell.

4/9/2015 DNO 42
Slide Blood Typing Group AB
The left hand of the slide contains anti-A which
reacts with the unknown cell.
The right hand side contains anti-B which reacts
with the unknown cell.

4/9/2015 DNO 43
Summary of Slide Typing
Anti-A Anti-B Blood Group

NEG NEG O

POS NEG A

NEG POS B

POS POS AB

4/9/2015 DNO 44
Distribution of Rh(D) Types
Population Rh(D) pos Rh(D) neg

Caucasian 86% 14%

African- 95% 5%
American
Oriental >99% <1%

4/9/2015 DNO 45
Significance of Rh(D)
80% of Rh(D) neg persons exposed to Rh(D) pos
blood will develop anti-D
Anti-D can also be stimulated by pregnancy with
an Rh(D) positive baby
• Sensitisation can be prevented by the use of anti- D
immunoglobulin, antenatally and post natally
Rh(D) neg females of childbearing potential
should never be given Rh(D) positive blood
products
4/9/2015 DNO 46
Inheritance
ABO & RH genes are not linked
ABO & Rh(D) type are inherited
independently
For example:
An A Rh(D) pos mother and a B Rh(D)
pos father
could have an O Rh(D) neg child

4/9/2015 DNO 47
Inheritance of ABO and Rh(D)

Mother Father

Group A AO Group B BO
Rh(D) pos Dd Rh(D) pos Dd

Group A AO Group B BO Group O OO

Rh(D) pos Dd Rh(D) pos Dd Rh(D) neg dd

4/9/2015 DNO 48
Question IS What Is your Blood Group

4/9/2015 DNO 49