Genotyping And Gene Polymorphism Of Human ABO Blood Group,HAB Secretor Status And MN Blood Group

Aim: To set up the methods of polymerase chain reaction-sequence specific primers (PCR-SSP) for human ABO blood group, HAB secretor status and MN blood group genotyping and study the polymorphism of ABO blood group genes, HAB secretor status genes in part of Han population in Chinese and MN blood group genes in random blood donors and study the application of ABO genotyping in clinical transfusion and genetics. Methods: the sample DNA was extracted from periodical blood by rapid-salt-method or by traditional phenol-chloroform method from amniotic fluid, and the PCR-SSP based methods were introduced to designate the genotypes of ABO blood group, HAB secretor status and MN blood group. Results: Seven selected DNA samples previously known genotypes were detected , proving the reliability of the genotyping for ABO blood group. ABO genes frequencies of 104 healthy and unrelated Han individuals were 0.1538 for A? 0.0962 for A2, 0.2452 for B, 0.5048 for 0?and O2was not found (X2=6.7323, P>0.25, the data fit Hardy-Weinberg distribution), having concordance with the serological result. Three patients with unclear ABQ blood group by the routine serological method, were readily detected by our genotyping method as B0? A20?and A20?respectively. Two cases of antigens low expression caused by leukemia chemotherapy were designated as B0抋nd A 態. A case of disputed paternity, which was not exclusive by ABO, MN. Rh and HLA- A,B,DR,DQ systems, proved to be no paternity by ABO genotyping. An amniotic fluid specimen from antepartum diagnosis was detected as A2B. Three reported ABO subgroup Bx, AB2, and A2B by serological method, were designated as 0 A20?and B0?respectively by genotyping. The frequencies of HAB secretor gene (Se) and non- secretor gene(se) were 0.9808 and 0.0192 respectively by three pairs of specific primers (X2~Z0.042l, >0.75, the data fit Hardy-Weinberg distribution). Compared with the reported serological results, the frequency of se in this study is 3ignificantly decreased. Thus we selected four individuals previously detected as non- secretor phenotypes by serological method to be detected by this genotyping method, and all were designated as Se/Se, which suggests that other mutations may exist in these samples. Compared with the serological method, PCR-SSP for MN blood group has concordance with the serological phenotypes. The frequency of M and N in 103 Chinese donors is 0.4806 and 0.5194 respectively~ which has not statistically significant deviation from Hardy-Weinberg equilibrium. Conclusion: The results demonstrate that PCR- SSP used in this study. is a convenient and reliable method for ABO and MN genotyping, and should be of great value in clinical and forensic application. We also deduce that the mutation of G447A and G757A can not cover all the se genes in Chinese Han population.