Molecular Biology Study And Application On ABO Genotype In Beijing

Objective To establish a diagnostic method for ABO genotyping. To study the ABO distribution of genotype in Beijing Han population. To provide a simple and reliable technique for safe transfusion and identification of complicated blood group. To develop a technology for ABO genetics study on rare blood group. Methods Develop an ABO genotyping method by using multiplex-PCR-RFLP and PCR-SSP techniques. Study molecular genetics on rare blood group by DNA cloning and sequencing technique. Apply microfluidic chip analytical systems to diagnose genotyping on clinical samples with serological problems. Results We have developed a set of methods, which are stable and accurate for ABO genotyping. Our study showed that A₁₀₂, O₁ and B allele were more common genes in Beijing Han individuals. And A₁₀₂ was predominant allele in the phenotype A group in this population. Three O₂ alleles were found and no A₂₀₁ allele was found while gene frequencies investigation was performed. We didn't discover the A₁₀₁A₁₀₁, A₁₀₁O₂, A₁₀₂O₂, BO₂ and O₂O₂ in this population. We have detected a rare A201 sample that contains anti-A₁. Using microfluidic chip analytical systems has identified a sample with decreased B antigen, which caused by CML. Our DNA cloning and sequencing technique found a rare B(A) sample with nt700(C→G) mutation. Conclusion Our study established a ABO genotyping diagnostic system by combining the microfluidic chip analytical systems with Multiplex-PCR-RFLP technique. Through its application on ABO genotyping for clinical difficult cases, this diagnostic system have done excellent job. The DNA cloning and sequencing technique for ABO gene provides a reliable guarantee on ABO genotyping and a direct method for further study on molecular genetics of rare blood group.