In this study, we molecularly detect the Jka/Jkb polymorphism in Taiwanese, Thais, and Filipinos with PCR-restriction enzyme digestion. We designed the primers with mutagenic bases to create two restriction enzyme cutting sites (EcoN I and Xba I) simultaneously to confirm the genetic polymorphism of Jka/Jkb. One hundred and seven unrelated Taiwanese, 107 Thais foreign laborers, and 102 Filipino foreign laborers in Taiwan were analyzed. The frequency of Jka and Jkb is 0.51 and 0.49 in Taiwanese, 0.44 and 0.56 in Thais, and 0.45 and 0.55 in Filipinos, respectively. The molecular analysis of Jka/Jkb polymorphism is simple, accurate and could confirm the uncertain results obtained by serological typing. The method could be used for predicting fetal Kidd genotype from amniocyte DNA, and to avoid the hemolytic disease of newborns caused by Jk antibody.