Experiment 6&7: Forward typing and Reverse typing Causes of False Positive

I. Blood collection 1. Drying of reagent

 Each group should have 2 known A 2. Dust/bacteria contamination
and 2 known B 3. Over centrifugation
 Known A – discard supernatant,
wash 3x and prepare 5% RBC Causes of False Negative
 Known B – discard supernatant, 1. Failure to add reagent
wash 3x and prepare 5% RBC 2. Expired reagents
 These two will be used as known 3. Undercentrifugation
cells.
𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑒
 Reagent: % RBC = Procedure
𝑇𝑉
𝟓% 𝒙
 = 𝟒𝒎𝑳 1. Slide Method
𝟏𝟎𝟎
× = 4 ( 0.05)
Prick method
× = 0.2 mL x 1000 = 200 uL
 Collect blood from each other and Saw
confirm blood type Mix with different
1. Serum – save to use in applicator sticks, tilt
A B back and forth for 2
REVERSE TYPING minutes, observe
2. RBCs – wash and prepare 5% Patient blood 1 drop 1 drop agglutination
RBCs Result:
Anti-A antisera 1 drop -------- + = agglutination
Landsteiner’s Rule: 0 = no agglutination
Anti-B antisera ------- 1 drop

 “Under normal conditions in the ABO blood

group system, a person does not have
antibodies towards his own antigens. A person 2. Tube Method
will only have antibodies to the antigen he lacks FORWARD TYPING REVERSE TYPING
(naturally-occurring or immune antibodies)” Anti A Anti B A cells B cells
Anti-A antisera 1 drop - - -
ABH Blood Group System Anti-B antisera - 1 drop - -
Patients 5% 1 drop 1 drop - -
Blood Types Antigens Antibodies washed RBCs
A AAH Known A cells - - 1 drop -
Anti-B Known B cells - - - 1 drop
Patients serum - - 2 2
B BBH drops drops
Anti-A

AB ABH
No Antibodies Centrifuge for 15 seconds under medium
speed, gently dislodge and grade
agglutination
O H Anti-A,Anti-B
(only forming anti Result: 4+,3+,2+.1+,0
A,B if exposed)
Bombay Anti-
A,AntiB,Anti-H
(anti-A,B only
formed precious
exposure)
Para-Bombay ABH Anti-
A,AntiB,Anti-
H/secretions
may have ABH
Ags