Rapid DNA crossmatch analysis of HLA class II genotypic polymorphisms by temperature gradient gel electrophoresis in unrelated bone marrow donor selection

Eur J Immunogenet. 1994 Oct;21(5):313-24. doi: 10.1111/j.1744-313x.1994.tb00202.x.

Abstract

The present paper demonstrates the usefulness of the temperature gradient gel electrophoresis (TGGE) as a precise method for the rapid identification of HLA-DR full matched donors by molecular crossmatch analysis. The resulting TGGE fingerprints are highly diverse for different HLA-DR genotypes and identical for samples with identical HLA-DR type, as shown for a panel of 25 randomly selected PCR-SSO pretyped DNA samples. In addition, TGGE analysis of a panel of homozygous typing cell lines (HTCs) established, that even subtle differences in the DR4 subtypes are detectable by TGGE crossmatch analysis. Furthermore, TGGE crossmatch analysis of unrelated donor/patient pairs demonstrated, that only HLA-DR identical patient/donor combinations showed identical TGGE patterns. Thus crossmatch analysis by TGGE presents a novel basis for a rapid and safe unrelated bone marrow donor selection strategy. Only HLA class I identical donors which also show HLA-DR identity in the TGGE crossmatch test are selected for the final confirmation of the HLA class II genes by a suitable DNA subtyping method.

MeSH terms

  • Alleles
  • Bone Marrow Transplantation*
  • DNA Fingerprinting*
  • Electrophoresis, Polyacrylamide Gel / methods*
  • Genotype
  • HLA-DR Antigens / genetics*
  • Histocompatibility Testing / methods*
  • Humans
  • Polymerase Chain Reaction
  • Polymorphism, Genetic
  • Protein Denaturation
  • Temperature
  • Tissue Donors
  • Tissue and Organ Procurement*

Substances

  • HLA-DR Antigens