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Polymerase chain reaction (PCR)/single-strand conformation polymorphism (SSCP) analysis of the human HLA-DQB regions.

Genetic diagnosis of 13 alleles of HLA-DQB1 (0501, 0502, 5031, 5032, 0601, 0602, 0604, 0201, 0301, 0302, 3032, 0401 and 0402) from 65 human DNA samples was achieved by applying single-strand conformation polymorphism (SSCP) analysis to DNA fragments amplified by the polymerase chain reaction (PCR) using a convenient primer set for DQB1 (recommendation of the International Histocompatibility Workshop, 1991). Differences between strand images (narrow/distinct or broad/diffuse) from the individual alleles and their electrophoretic mobilities are regarded as criteria for confirming the genetic diagnosis of DQB1 alleles. This primer set amplifies not only DNA fragments belonging to DQB1, but also to DQB2, and classification of 3 phenotypes (1.1, 1.2 and 1.1/1.2) in the presence of two alleles at the latter locus was suggested. Consequently, PCR/SSCP of DNA amplified by this primer enables classification of the phenotypes, at least under our experimental conditions, into 3 x 91 groups. Two advantages of SSCP analysis over VNTR with regard to the use of amplified DNA in forensic practice are described.