ABO BLOOD GROUPING Forward Typing Principle:

Resuspend the cells w/ gentle agitation & examine macroscopically for agglu Interpretation: same as forward slide method + detection of weak subgrps of A* ABO grps represent Ags on RBC surface + Ab in o Note: vitro clumping of RBCs expressing Ag Dont rely on color of dyes to ID rgt antisera. All o Ab & corresponding Ag not present in same blood tubes must be properly labeled specimen Dont perform tests at T > than RT (20-24 C) o Det. of ABO grp. for pretransfusion studies of Pxs & Perform observations of agglu w/ well-lighted BG, donors; specialized (obstetric) cases not a warm view box Specimen: Record results immed. After obs. o No special prep of the Px needed Contaminated blood specimen, rgts or supplieso (+) ID of Px when specimen is collected interfere w/ results st Reverse Typing Label specimen at bedside. (Pxs 1 & last name, Principle: date collected, Pxs hospital ID no., time of o Confirm ABO blood grping collection, phlebos initials on the request form) o Based on the presence or absence of Abs anti-A & o Blood drawn by aseptic technique & tested right anti-B in serum away o Crosscheck for forward typing o Draw 5-7 ml of blood in plain (red top) or in an EDTA o Not performed on specimens from newborn & very (lavender top) vacutainer tube. Newborn infant young infants lack of synthesized Igs samples maybe collected in pediatric blood specimen Rgts, Supplies & Equip: containers or as RBCs in normal saline o Rgt RBCs: A1 & B (A2 optional); Known A & B cells Rgts, Supplies & Equipment o 10 x 75 mm TTs o Commercial blood grping antisera (Anti-A, Anti-B, o Pipettes anti-A,B) o Lamp o 10 x 75 mm disposable TT (for TT method); ceramic o Centrifuge ring slides (slide method) o NSS QC: rgt RBCs tested daily w/ known antisera o Disposable pipets 4 5/8 plastic or Pasteur Procedure o High intensity lamp/ optical magnifying lens o Label TTs: A, B, O, P o Centrifuge o 2 drops test serum/plasma o 1 drop A1, B & O rgt RBCs*; P tube: Pxs own QC: Rgt antisera tested daily w/ RBCs of known washed RBC antigenicity *or freshly prep. 2-4% susp. of washed A, B & O Procedure: cells o Slide Method o Mix well & centrifuge TT for 15 s, 3500 rpm ID ceramic ring slide w/ Pxs name & no. o Resuspend cells by agitation & examine Label A, B, O, P macroscopically for agglu 1 Drop each of anti-A, anti-B, grp O, Pxs serum Abs (Pxs autoAb) A cells B cells O cells Blood Grp present Small amt of blood (not > than of the vol. of + Anti-B A serum used; ratio = 2 serum: 1 blood) next to + Anti-A B each drop of antiserum w/in circ. Use wooden applicator stick or pipet None AB Finger blood, venous or AC blood may be used Anti-A & + + O Mix w/ separate applicator sticks. Tilt slide Anti-B backward & forward & observe for macro agglu. o Hemolyzed specimen unsuitable for this test; Dont warm slide! Agglu will occur w/in a few s. testing should be at RT or colder exc. in the case of weak Ags of Grp. A & B cells Max obs. Time: 2 mins. Interpretation Ags Anti-A Anti-B Grp O Blood Grp present + + A A + + B B + + + A, B AB None O C weak subgrps + factor of A* o Test Tube Method More sensitive bec of: Centrifugation Use of RC suspension prevent false (-) Prep. 2-4% suspension of Pxs RBCs in NSS (washing: 2x) 4 TTs: A, B, O, P 1 drop anti-A, anti-B, grp O, Pxs serum

Using disposable pipette, 1 drop RC susp each (1

3); last tube: Pxs own washed RBC Mix well & centrifuge TTs for 15 s at 3400 rpm

st