ABO Genotyping Following a Single PCR Amplification

Using primers designed by Lee and Chang, 200 base-pair (bp) fragment of ABO locus was amplified by PCR, which spans the site of the single nucleotide deletion associated with O allele. O allele could be identified by Kpn I digestion of the PCR product as reported. A and B alleles were also distinguishable by Mae II digestion of the product. Thus restriction digestion by Kpn I and Mae II could genotype ABO blood group following the single amplification. The nucleotide substitution in the 200-bp product between A and B alleles was also found in O allele, resulting in 2 different suballeles OA and OG. The single-strand conformational polymorphism of the PCR product was also investigated for ABO genotyping following the single amplification.