A rapid HLA-DRB1*04 subtyping method using PCR and DNA heteroduplex generators.
作者： N. A. P. Wood , J. Evans , J. L. Bidwell , J. L. K. Wee , A. A. Oei
DOI: 10.1111/J.1399-0039.1996.TB02555.X
关键词:
摘要: We describe here a rapid polymerase chain reaction (PCR)-based method for the identification of HLA-DRB1*0401-*0412 alleles. The is based on generation specific DNA heteroduplex patterns between PCR products derived from selective group-specific amplification various DRB1*04 alleles and two synthetic generator (DHG) molecules following non-denaturing polyacrylamide minigel electrophoresis. One DHG was designed to detect DRB1*0401, *0405, *0407, *0408, *0409 alleles, whilst other DRB1*0402, *0403, *0404, *0406, *0410, *0411, *0412 Characteristic were obtained all tested both in homozygous heterozygous situations. Both PCR-SSP (sequence-specific primer) typing performed 41 positive DNAs Singaporean Chinese blood donors complete concordance results obtained. HLA-DRB1*0403, *0406 found account 95% population studied. technique described technically simple since it essentially involves only amplifications per individual subtyping. particularly useful resolving combinations which are indistinguishable by PCR-SSO oligonucleotide) or
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