ABO GENOTYPING BY MUTAGENICALLY SEPARATED POLYMERASE CHAIN-REACTION

ABO blood groups were determined by the mutagenically separated polyme rase chain reaction (MS-PCR). The products from two sets of PCR reacti ons using the same program for the nucleotides at positions 261 and 70 3 from cDNA at the ABO locus were used to distinguish A, B and O allel es. Two forward mutagenic allele-specific primers of different lengths for the ABO polymorphic site were paired with the same reverse primer in each PCR reaction. The 216 bp fragment of the PCR products for the 261th nucleotide was A or B allele-specific and the 195 bp fragment w as O allele-specific. The 126 bp fragment of the PCR products for the 703th nucleotide was B allele-specific and the 106 bp fragment was A o r O allele-specific. The ABO genotypes were determined by the intersec tion of the predicted alleles from these two PCR reactions. The PCR pr oducts were obtained using 10 ng of DNA in 50 mu L of PCR reaction mix ture, and electrophoresed in 4% agarose gel. In this study, 265 ABO-ph enotype known samples (A: 31, B: 48, AB: 6 and O: 180) in Chinese were used. The results of ABO genotypes were AA: 1, AO: 30, BB: 2, BO: 46, AB: 6 and OO: 180. These results were confirmed by the PCR-RFLP ABO g enotyping method. This technique is a simple, rapid, and reliable meth od for ABO genotyping.