A Simplified ABO Genotyping by Allele-Specific Polymerase Chain Reaction

Background : Genotyping of ABO gene could be more informative and valuable than serological typing in some situations such as the resolution for ABO discrepancy between the cell typing and serum typing and determination of A and B subgroups. We developed a simple allele-specific polymerase chain reaction (AS-PCR) method without the use of any restriction enzymes to detect the A, B, O, and cis-AB alleles for Koreans. Methods : An AS-PCR was designed with amplification refractory mutation system (ARMS) at nt (nucleotide) 261 (exon 6) and at nt 526, 803 (exon 7) of ABO gene to detect specific nucleotide sequence differences between the ABO alleles. We tested for ABO genotyping 60 DNA samples previously tested by PCR-RFLP and stored at . These samples had been obtained from blood donors recruited at the Gwangju-Chonnam Red Cross Blood Center between July 2002 and February 2003. Results : With our new PCR method, the genotypes of the 60 samples were found to be A/O (n=10), A/A (n=5), B/O (n=10), B/B (n=5), O/O (n=10), cis-AB/A (n=5), cis-AB/B (n=5), and cis-AB/O (n=10), which were the same results obtained previously with PCR-RFLP. Conclusions : Our AS-PCR is a simple and accurate method for the detection of A, B, O, and cis-AB alleles for Koreans.