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We have developed an allele-specific polymerase chain reaction (AS-PCR) system to detect HLA-DQB1*0601 allele by using DNA amplified by a conventional PCR system. In AS-PCR, primers were designed to anneal to regions specific to HLA-DQB1*0601 and the third allele (DQB1*0602,*0603,*0604) of the human leukocyte antigen (HLA)-DQB1 genes. After the first-round PCR, the second-round PCR was performed using the allele-specific primer, and the products were further amplified. This system was found to be highly specific for the HLA-DQB1*0601 allele, even when the HLA-DQB1*0602,*0603,*0604 alleles were also present at a low frequency. With the method, HLA-DQB1*0601 allele was detected in 22 be359ba680
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