Red cell phenotyping using hexadimethrine bromide (Polybrene) in a microplate system

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Abstract
To provide rapidly phenotyped units of blood, the hexadimethrine bromide (Polybrene) technique was adapted to microplate technology. Pilot samples from 282 donor U were phenotyped for antigens in the Rh, Kidd, Kell, Duffy and Ss systems with a standard tube-testing method and a Polybrene-microplate (P-MP) technique. Diluted antisera and a 1% red cell suspension were used to give P-MP reactions that were accurate and easy to interpret. One microplate, containing 96 tests, was prepared and read within 15 min for P-MP tests yielding direct agglutination (Rh, Jkb, Fya, Fyb), or within 19 min for P-MP tests requiring an antiglobulin phase (Jka, K, S, s). No false-positive results were found. No false-negative reactions were found in typing for Rh, K, S, s or Jka antigens. Phenotyping for Jkb and Duffy antigens gave a false-negative rate of less than 0.015. Considerable savings in reagents were obtained through microplate miniaturization and through the enhancement of apparent antibody avidity in the Polybrene/low ionic medium which permitted dilution of reagent antisera. The P-MP technique affords a rapid, accurate, simple, and inexpensive means of phenotyping large numbers of donor U.

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