Quantitation of ABO mixed-cell populations by a peroxidase-anti-peroxidase immunoenzyme method

The peroxidase-anti-peroxidase immunoenzyme method was applied to red blood cell ghosts for the detection of ABO mixed-red blood cell populations. Red blood cell ghosts produced by acid-glycine lysis eliminated previous problems due to distorted red blood cell and hemoglobin-associated peroxidase activity. Anti-A,B was used to discriminate between various mixtures of test cells in group O cells. The positive-stained minor population of test cells stained dark brown following application of the peroxidase-anti-peroxidase method and the nonreactive group O cells were counterstained light blue with Coomassie brilliant blue which facilitated quantitation. The expected and observed numbers of positive-stained cells per 1000 cells were not significantly different as computed by X2. As low as a 0.5 percent minor population in an ABO red blood cell mixture could be accurately quantitated by this method.