Establishment of Mixed Passive Hemagglutination Assay Using In-House Anti-Rh(D) Sensitized Human O RBCs for the Detection of Granulocyte Specific Antibodies / 대한수혈학회지
The in-house indicatorcells for MPHA were made by sensitizing human Rh(D) positive O RBCs with humanIgG anti-Rh(D) (DiaMed AG, Switzerland) and then combining with AHG anti-IgG (Immucor Inc., USA). To determine the optimal conditions, various combinations of anti-Rh(D) IgG sensitization strengths of indicatorcells, microwell coated antigens (intact granulocyte vs. extracted granulocyte) and reaction conditions were compared.
RESULTS:
The best test conditions for MPHA were as follows optimal results were obtained with the anti-Rh(D) sensitization dilutions of 1/64-1/192 and the reaction condition of 4 hours incubation at room temperature in humid chamber. Extracted granulocytes coated at the plate showed better results than intact granulocytes. HLA antigens were completely removed from extracted granulocyteantigens after acidified chloroquinetreatment.