title: Rapid DNA crossmatch analysis of HLA class II genotypic polymorphisms by temperature gradient gel electrophoresis in unrelated bone marrow donor selection.
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.
authors.affiliation: Bone Marrow Donor Center (with Transplantation Immunology and Stem Cell Banks), Heinrich-Heine University, Dusseldorf, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Uhrberg
authors.forename: J
authors.initials: J
authors.lastname: Enczmann
authors.forename: P
authors.initials: P
authors.lastname: Wernet
mesh_terms: D000483:Alleles; D016026:Bone Marrow Transplantation; D016172:DNA Fingerprinting; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005838:Genotype; D006684:HLA-DR Antigens / Q000235:genetics*; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D011489:Protein Denaturation; D013696:Temperature; D014019:Tissue Donors; D009927:Tissue and Organ Procurement
chemical_list: D006684:HLA-DR Antigens