Resolution of cis-trans ambiguities between HLA-DRB1 alleles using single-strand conformation polymorphisms and sequencing.
DNA-based typing of HLA alleles occasionally results in the inability to assign a specific allele because of ambiguity in associating two or more polymorphisms to the same or to alternate homologs (cis/trans ambiguity). Since most individuals are heterozygous at a given HLA locus, the highest level of confidence in definition is obtained when the alleles are tested in isolation. By using single-strand conformation polymorphisms (SSCP) to separate heterozygous HLA-DRB1 alleles, followed by sequencing of separated conformers, we have achieved resolution of previously ambiguous assignments without the need for additional probes or primers.
Paul
P
P
Allogen Laboratories, The Cleveland Clinic Foundation, Cleveland, Ohio, USA. pp@tt.ccf.org
Thomas
D
D
Kawczak
P
P
Good
D
D
Cook
D J
DJ
Ball
E J
EJ
D000483:Alleles; D000595:Amino Acid Sequence; D001483:Base Sequence; D014644:Genetic Variation / Q000276:immunology; D006684:HLA-DR Antigens / Q000032:analysis* / Q000235:genetics*; D059811:HLA-DRB1 Chains; D006650:Histocompatibility Testing / Q000379:methods; D006801:Humans; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D018807:Polymorphism, Single-Stranded Conformational; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA / Q000379:methods; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid
D006684:HLA-DR Antigens; D059811:HLA-DRB1 Chains