ABO BLOOD GROUPING AND RH TYPING

The blood contains RBCs with different types of antigens on its

surface, while the plasma contains different antibodies.
There are more than 300 blood group systems but ABO and
rhesus (Rh) systems are of importance from clinical point of view
because A, B & Rh D antigens are most immunogenic. The ABO
system of grouping concentrates on the type of antibody present
in the plasma and the antigen present in the RBCs. The gene for A
& B antigen is present on chromosome 9 and for Rh antigen on
chromosome1.ABO system was discovered by Landsteiner in
1900.

ABO SYSTEM
Principle:-
The red cells contain different types of antigens (agglutinogen),
while plasma contains antibodies (agglutinins). In order to
determine the blood group of a subject, the red cells are allowed
to react with a sera containing known antibody (agglutinin).

FORWARD (CELL) GROUPING:

Red cells are tested for presence of antigens by employing known
antisera.
REVERSE (SERUM) GROUPING:
Serum is tested for presence of antibodies by employing known
red cell reagent.

Methods for ABO Grouping:-

1. Slide method
2. Tube method

1. Slide Method:-
Take a clean glass slide.
Divide one slide into two halves with a glass marking pencil and
mark these areas as A and B.
Place a drop of serum anti-A (blue) on the slide in area marked A
and a drop of serum anti-B (yellow) in the area marked B.

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 Make a finger prick with a disposable needle after cleaning the
area.
Place a drop of blood near anti-A and anti-B serum and mix them
with a stick or with the end of a glass slide.
Wait for 5 minutes and look for agglutination.
Observations:-
If any agglutination occurs, it is visible with naked eyes as dark
reddish clumps of different sizes. If agglutination is minimal it can
be confirmed by examining it under a microscope.

Note:-
Red blood cells from person with blood group O contain no
antigen; so no agglutination occurs and thus such a person is a
universal donor.
Red blood cells from persons with blood group AB contain both A
and B antigens, so agglutination occurs in both as no antibodies
are present in their serum and thus such individual is a universal
recipient.
RHESUS (Rh) SYSTEM
Rh blood group system was first reported by Landsteiner and
Weiner in 1940. In contrast to ABO system, Rh antigens are
present on red blood cells only and Rh antibodies develop only in
response to a known stimulus (e.g. blood transfusion, or after first
pregnancy). Rh factor is present in 85-95% of human beings.

Methods for Rh Grouping:-

1. Slide method
2. Tube method
1. Slide Method:-
Take a clean glass slide.
Place a drop of anti-D serum on the slide.
Place a drop of blood near anti-D serum and mix them as for ABO
grouping.
Wait for 5 minutes and see for agglutination

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Note:-
Rh positive subjects have Rh antigen on their red blood cells but
no Rh antibody in their serum.
Rh negative subjects have neither Rh antigen on their red blood
cells nor Rh antibody in their serum. The most common Rh
antigen is D.

Incidence of ABO Blood Groups in India

A = 22-27% ,B = 31-33% , O = 34-40% (most common) , AB = 5-8%
IMPORTANCE OF BLOOD GROUPING:-
1. Blood transfusion
2. Haemolytic disease of new born (Erythroblastosis Foetalis)
3. Paternity disputes
It is possible to disprove parentage but impossible to prove
parenthood.
4. Medico-legal use
In criminal cases, whether a stain is blood or not, and its blood
group can be detected.
5. Used in immunology, genetics and anthropology
6. Susceptibility to various diseases
Persons with blood group O are more susceptible to peptic ulcer
while persons with blood group A are more susceptible to gastric
cancer.

LANDSTEINER’S LAW:
Anti-A and/or Anti-B antibodies are always present in plasma of
individuals who lack corresponding antigen on their red cells.

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 ANTIGENS AND ANTIBODIES IN ABO GROUPS

GROUP ANTIGEN ON RBC ANTIBODY IN SERUM

A A ANTI-B

B B ANTI-A

AB A&B ----

O ---- BOTH ANTI-A & ANTI-B

ABO GROUPING

ANTI – A ANTI – B INTERPRETATION

( BLUE COLOR ) ( YELLOW COLOR )

AGGLUTINATION NO AGGLUTINATION A GROUP

NO AGGLUTINATION AGGLUTINATION B GROUP

AGGLUTINATION AGGLUTINATION AB GROUP

NO AGGLUTINATION NO AGGLUTINATION O GROUP

RH TYPING
ANTI – Rh INTERPRETATION

( COLORLESS )

AGGLUTINATION +++ ( POSITIVE )

NO AGGLUTINATION --- ( NEGATIVE )