Rapid DNA typing utilizing immobilized oligonucleotide probe and a nonradioactive detection system application to HLA-DR typing of the Japanese population

Abstract

We established a rapid and simple method of HLA-DR genotyping, and applied it for analysis of the Japanese population. Our method includes rapid preparation of DNA samples from buccal mucosa, incorporation of biotin-dATP into DRB genes during amplification by the polymerase chain reaction, hybridization with sequence-specific oligonucleotide (SSO) probes immobilized on nylon membranes via poly (dT) tails, and detection of the hybridization signal as chemiluminescence. We carried out DR typing of 30 Japanese donors using 20 different immobilized SSO probes, and obtained unambiguous typing signals showing perfect correlation with their serologic DR types. The genotyping also enabled us to identify several DR types unique to the Japanese population, such as DRw12b (DRB1 ∗1202), DRw14c (DRB1 ∗1405), and serology blank type, DR‘JX6’ (DRB1 ∗1403). The method presented here would be suitable for routine DR typing in tissue-typing laboratories.