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II Chapter-16 II 6
Bl oo d Grouping & R h Typing
Introduction
Blo od transfusion is a tremendous invent
ion and life saving procedure in many
sev ere blo od los s and severe anemia. Ho .
.
has me dic o-l ega l importance also.
we ver blo od can only be giv en after blood groucase
.
s of
ping It
·
Principle
Re d cel l me mb ran es contain antigens kno
wn as isoantigens (agglutinogens) in their
me mb ran es, wh ere as pla sm a contains cell
antibodies called as agglutinins. For the
gro upi ng, per son 's red cells are made to purpose of blood
react with available antisera wh ich contain
bod ies or agg luti nin s. Th en the slide is known anti-
observed under the Mi cro sco pe to detect
abs enc e of hem oly sis of red cells which the presence or
occurs as a result of antigen- antibody reac
tion.
Re qu ire me nts
1. Pri cki ng apparatus. (Lancet/ Pricking nee
dle, cot ton swab, sprit etc.)
2. 5 ml tes t tub es.
3. Gla ss slides.
4. 1% sod ium citrate in normal saline.
5. An ti-A ser um ( Contains anti-A antibodie
s).
6. An ti-B -se rum ( Contains anti-B antibodie
s).
7. An ti-D -se rum (Co nta ins anti-Rh antibo
dies).
Procedure
1. Us ing a ma rke r div ide eac h slides in to
two equ al halves.
2. Le ft hal f is tak en as a tes t sid e and rig ht
as con tro l sid e or vic e versa.
3. To tal 4 slid es are tak en for the procedure
. 1 eac h for anti-A, anti-B , and anti-D •And 4th
slid e is ma rke d as 'S' i.e. no ant ise rum
will be add ed to this slide.
4. Pla ce 8-1 O dro ps of sal ine sol utio n in
the Cen tre of the slide 'S'.
5. Pre par ati on of Re d cel l sus pe~ sio. . . 1· .
ins tea d of add ing blo od dro ps directly
n - A suspension _ of red cel ls i.n s~ me is prepared
with
fro m fin ger pnc k bec aus e dilution of bloo
d
 . g & Rh typing /- · ,.. .
11plP
" ~ } ....• . ..,,, ~
~ Gro .
~ onits easy detection of agglutination and 1 . • 14 I
Jji1epe . 1 d. h . a so it promote h .
s~ . which are mvo ve mt e mterference of 1utination. . . st e ehmination of I
factois . . . agg Pasma
aseptic conditions Th
f. ger pnck. under
5ml f a1i . . en add 2 drops f bl
Get a in by addmg o s ne usmg stick/ tooth pnc .k , o ood to the slide 'S,
.xit .
'tndnu
' unation of Blood groups- Place I drop of Ant· A
pctet11 . 1- serum on left h Jf
1 drop of Ant1-B over left half of the 2nd test tub d a of the Ist test
tvhe, d t e an 1drop of anti-Dover left half
ot the 3r tes .
to Left and right h f
F1·omthe slide 'S' Put one drop. of solution each a1ves o 3 test tubes.
Mix the antisera and blood - saline solution using the stick/ tooth prick.
Mixing can also be done by blowing on them.
~.
Wait for 8-10 minutes and then observe both the control and test sides of slides first with
I. the naked eye to detect the agglutination.

11.
Then confirm the results using microscope under the Low power.
\l Test side has to be compared with
the control side.
Precautions
I. 3separate sticks should be used for mixing.
l. While mixing care shovld be taken to stop the mixing of test solution and control solution.

Anti-A Anti- B Anti-D

/
-

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l. Slides should be dust free and grease free. t the error in results.
4. h . h t fail to preven
iviarking of slide should be done wit ou
Observations and results It appears as the isolated, dark- red
If h . . . . . 1 t0 the naked eye.
tnas t ere 1s agglutination 1t 1s v1s1b e
ses of .
Vanous sizes and shapes.
 Practical Sha rirK riya '
142
Clinical importance ~
Indications for ABO grouping
ABO grouping is required for all of the following individuals -
► Blood Donors_ Since it can be life threatening to give the wrong ABO
group to the '
patient.
► Transfusion recipients - Since we need to know the donor blood is ABO
comp t'b
a 1 le.
► Transplant Candidates and Donors - ABO antigens are found in other
tissues as Well
Therefore the transplant candidates and donors must be compatible.
·
► Prenatal Patients-To determine whether the mothers may have babies who
are suffering
fromABO-HDN.
► Newborns (sometimes) If the baby is demonstrating symptoms of Hemolytic
Disease of
the Newborn, the ABO group needs to be determined along wiG1 ru1 and
others.
► Paternity testing since the inheritance of the ABO Blood Group system is
very specific,
this serves as one of the first methods to determine the paternity issues.
Note : ABO antibodies are of major clinical significance for two reaso
ns - They are
naturally occurring and are found universally and they are highly reactive.
Transfusion reactions
► The routine practice of blood typin g and c_,~ ·. / -': '; . z: >_·; 1ing blood products shoul
-.,_·1· 1'
d prevent
adverse transfusion reactions caused b:-, /\.; "-·· · ;c~\. d xKlies.
► If a recipient who has blood group O is tranE,fi.1.:; ~d with non-group ORB Cs,
the naturally
occurring anti-A and anti-Bin the recipient's serurn binds to their correspondi
ng antigens
on the transfused RBCs. These antibodies fix complement and cause rapid
intravascular
hemolysis, triggering an acute hemolytic transfusion reaction that can cause
disseminated
intravascular coagulation, shock, acute renal failure, and death.
► Most cases of hemolytic disease of the newborn (HDN) that arise from an
ABO incom-
patibility require no treatment. Cases of severe hemolysis that require excha
nge tranSfu-
sions are less common, and fetal hydrops is rare.
► HON caused by ABO antibodies occurs almost exclusively in infants of blood
groupA~r
B who are born to group O moth ers. This is becau se the anti-A and 1
anti-B formed ~
group O individual s tend to be of the IgG type (and therefore can cross
the placenta ~
whereas the anti-A and anti-B found in the serum of group B and Am . d'1 1.d als respec
v u '
tively, tends to be of the IgM type. Although uncommon, cases of HDN
have been re-
ported in infants born to mothers with blood groupA2 and blood group B.
 143
d Grouping & Rh Typing
9100
adult
~l{DN tends to be relatively mild in nature mainly because fetal RBCs don't express
ens can
'r levels of A and B antigens. However, the strength of fetal ABO blood group antig
be unpre-
v'af.Y, and therefore the degree of hemolysis and hence the severity of HDN can
develop-
dictable. Early studies suggested that the race of a neonate was a risk factor for
se that re-
ing ABO HDN. However, later studies showed that the prevalence of disea
Caucasian
quired treatment did not differ significantly among Asian, Black, Hispanic, and
infants.

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