NILAI UNIVERSITY COLLEGE

DIPLOMA IN MEDICAL LABORATORY TECHNOLOGY

MPAD 1221
PRACTICAL 2
Name : Tan Hui Jiang (heidi)
Date : 19.02.2009
Objectie
To determine ABO grouping of blood sample (e!erse "rouping)
P!i"ci#$e
Tube and tile method ma# be used for re!erse grouping of B$s. e!erse grouping is
performed to %onfirm the presen%e of e&pe%ted naturall# o%%urring antibodies in the
serum or plasma. eagent B$s demonstrating the strongest antigeni% ma'eup for A and
B are used (A
1
and B %ells).
Rea%e"t&
A %ells( B %ells and O %ells

Sam#$e
)erum or plasma
P!'ce()!e
Tube method
1. 2 drops of the serum or plasma are pla%ed to be tested in ea%h of t*o appropriatel#
labeled tubes (A
1
and B %ells).
2. 1 drop of reagent A
1
%ells is pla%ed in the tube mar'ed +A
1
%ells, and one drop of
reagent B %ells is pla%ed in the tube mar'ed +B %ells,.
-. .i&ed thoroughl# and %entrifuge at 1000g for 1/020 se%onds.
1. The B$s are gentl# resuspended *hile the agglutination is obser!ed and the results
are re%orded.
Re&)$t
Patie"t A ce$$& B ce$$& O ce$$& B$''( G!')#
1 0 2 0 A
2 2 0 0 B
- 2 2 0 O
1 0 0 0 AB
P!'ce()!e
Tile method
1. 1 drop of ea%h reagent to be tested (A %ells( B %ells or O %ells) is pla%ed on a %lean
glass slide or tile allo*ing appro&imatel# a 20 3 10 mm area for ea%h test.
2. 1 drop of *ell mi&ed serum or plasma is added to ea%h drop of reagent in step 1.
-. The reagents and serum or plasma are mi&ed thoroughl# using a %lean *ooden
appli%ator sti%' *hile spreading the mi&ture o!er an area appro&imatel# 20 3 10
mm.
1. The tile ba%' is gentl# tilted and forth for 2 minutes *hole agglutination is obser!ed
and the results are read and re%orded.
Re&)$t
Patie"t A ce$$& B ce$$& O ce$$& B$''( G!')#
1 0 2 0 A
2 2 0 0 B
- 2 2 0 O
1 0 0 0 AB
C'"c$)&i'"
Tile method is easier to be %arried out but tube method is more a%%urate. 4etermination
of ABO grouping of patient sample %an easil# done in lab( but there is a limit for
'no*ing the genot#pe. This method %an onl# sho* us the phenot#pe5blood group of the
patient.