[A study on HNA-1~-5 genotypes and frequencies in the blood donor population of jiangsu province].

Abstract

Objective To investigate the distribution characteristics of the genotypes and allele frequencies of HNA-1~-5 alleles in a blood donor population from Jiangsu Province. Methods Genomic DNA was extracted from 300 peripheral blood samples collected from healthy donors. All samples were genotyped using an optimized polymerase chain reaction-sequence specific primer (PCR-SSP) protocol. Randomly selected results were verified by direct sequencing, and genotype distributions were tested for conformity to Hardy-Weinberg equilibrium. Samples identified as potential HNA-1 null were confirmed by polymerase chain reaction sequencing-based typing (PCR-SBT) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results The allele frequencies for HNA-1a, HNA-1b, and HNA-1c were 58.6%, 41.4%, and 0%, respectively. Two identified HNA-1 null cases were confirmed to result from a complete deletion of the Fc gamma receptor IIIb (FCGR3B) gene. The allele frequency of the HNA-2-encoding CD177 gene was 100%, with no HNA-2 null cases detected due to the homozygous c.787 A>T (TT) mutation. The allele frequencies for HNA-3a, HNA-3b, HNA-4a, HNA-4b, HNA-5a, and HNA-5b were 63.0%, 37.0%, 100%, 0%, 88.8%, and 11.2%, respectively. No statistically significant differences in HNA allele frequencies were found between the Jiangsu population and those from Shanghai or Zhejiang. However, significant differences were observed when compared to the Guangzhou population specifically for the HNA-1a/1a (P=0.008), HNA-3a/3a (P=0.033), and HNA-3b/3b (P=0.022) genotypes. Conclusion This study establishes the HNA genotype and allele frequency profile for the Jiangsu blood donor population, providing essential baseline data for research into HNA-related alloimmunization. Furthermore, it optimizes a methodology suitable for large-scale HNA screening and establishes a comprehensive confirmatory testing protocol for HNA-1 null, thereby laying a technical foundation for subsequent research on this allele.