Simple ABO genotyping method using three polymorphic sites at the ABO locus

Kang, Sung Ha; Shin, Dong Hun; Cho, Hyun Chan; Lee, Kyu Man; Han, Kyou Sup

Abstract

BACKGROUND
Recent progress in the molecular biology of the ABO blood group system has recognized the molecular basis of the red cell antigens and has provided a genetic model for ABO polymorphism at the molecular level. Genotyping methods with the basis of this genetic model were tested for korean blood donors.
METHODS
Simple genotyping of the ABO blood group was performed in 253 healthy Korean blood donors by polymerase chain reaction-restriction enzyme length polymorphism (PCR-RFLP) and allele-specific PCR analysis of the only three polymorphic nucleotide positions (nps) 261, 526 and 803 of the ABO blood group gene. Two differen regions of the ABO gene, each of which contained a different polymorphic site (np 261 or 526) were amplified. Amplified products were digested with four restriction enzymes, two complementary pairs, Kpn I and Bst PI, and Bss HII and Ban I to analyze the np 261 and 526, respectively. The np 803 was analyzed by PCR with allele-specific primers. The PCR determined genotypes were compared with serologically determined phenotypes.
RESULTS
The results were consistent for all individuals and not different from the proposed genetic model. The ABO genotypes of 253 healthy Korean blood donors were13 AA, 64 AO, 11 BB, 57 BO, 81 OO and 27 AB.
CONCLUSIONS
The simple method used in this study may serve as a helpful tool for solving difficulties in serological ABO blood typing in the transfusion and forensic medicine, especially for cases with ABO discrepancy due to genetic variations. This method would be more efficient and informative for genotyping of the Korean population.