RHDalleles contributing to serologically weak D phenotypes in China: A single‐centre study over 10 years

BACKGROUND AND OBJECTIVES: In cases of serologically weak D phenotypes, RHD genotyping may identify discrepant serotyping results and protect the patient against allogeneic immunization. This study aimed to conduct a comprehensive analysis of weak D alleles in China. MATERIALS AND METHODS: We collected samples carrying weak D antigen during a 10-year period from 2005 to 2014. The intensity and epitopes of D were analysed serologically. Genomic DNA was extracted and used for RHD sequencing and heterozygote analysis. In particular, an in vitro expression method for functional verification of the rare and novel in-frame deletion mutation was developed and then combined with homologous modelling results for analysis. RESULTS: We studied a total of 283 weak D samples from volunteer blood donors and identified 45 RHD alleles among them, 11 of which were reported for the first time. Ten (3.5%) samples surprisingly carried DEL allelic variants and as many as 40 (14.1%) carried the wild-type RHD genotype. Combination of the results of functional experiments and in silico analysis suggested that the rare in-frame deletion mutation may reduce the expression of D antigen by affecting the RhD protein structure. CONCLUSIONS: This study provides an enhanced overview of the distribution characteristics of RHD alleles in Chinese subjects with serologically weak D. An in vitro method to predict the biological significance of variant RHD alleles was also provided. We found inconsistent genotyping and phenotypic results in some samples, indicating the existence of additional regulatory mechanisms.