HLA-DRB1*01 subtyping by heteroduplex analysis

Abstract: In this report we describe an alternative method for the identification of the four known HLA-DRB1*01 alleles, which is based on the generation of unique heteroduplex patterns between the different DRB1*01 alleles and a synthetic DNA heteroduplex generator (DHG) molecule. The method is technically simple, rapid and cost-effective, as it essentially involves only a single polymerase chain reaction (PCR) followed by polyacrylamide gel electrophoresis. This technique allows the rapid discrimination of the various known HLA-DRB1*01 subtypes, both in homozygous and heterozygous situations. We propose that this technology can potentially be applied to most HLA class I and class II subtyping.