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.