RAPID DNA CROSS-MATCH ANALYSIS OF HLA CLASS-II GENOTYPIC POLYMORPHISMS BY TEMPERATURE-GRADIENT GEL-ELECTROPHORESIS IN UNRELATED BONE-MARROWDONOR SELECTION

The present paper demonstrates the usefulness of the temperature gradi ent gel electrophoresis (TGGE) as a precise method for the rapid ident ification of HLA-DR full matched donors by molecular crossmatch analys is. The resulting TGGE fingerprints are highly diverse for different H LA-DR genotypes and identical for samples with identical HLA-DR type, as shown for a panel of 25 randomly selected PCR-SSO pretyped DNA samp les. In addition, TGGE analysis of a panel of homozygous typing cell l ines (HTCs) established, that even subtle differences in the DR4 subty pes are detectable by TGGE crossmatch analysis. Furthermore, TGGE cros smatch analysis of unrelated donor/patient pairs demonstrated, that on ly HLA-DR identical patient/donor combinations showed identical TGGE p atterns. Thus crossmatch analysis by TGGE presents a novel basis for a rapid and safe unrelated bone marrow donor selection strategy. Only H LA class I identical donors which also show HLA-DR identity in the TGG E crossmatch test are selected for the final confirmation of the HLA c lass II genes by a suitable DNA subtyping method.