A novel approach to simultaneous genotyping of human platelet antigen systems and human leucocyte antigen class I loci using PacBio long‐read sequencing

Abstract

Background and ObjectivesAccurate human leucocyte antigen (HLA) and human platelet antigen (HPA) typing is essential for establishing a blood platelet donor bank to deal with refractoriness in patients undergoing multiple platelet transfusions. Current methods, such as Sanger and next‐generation sequencing, encounter difficulties in haplotyping. Herein, the aim of this study was to establish a method for HLA and HPA typing based on the long read sequencing.Study Design and MethodsThe HPA and HLA class I genotypes of 268 platelet donors from the Taiyuan Blood Center, China were identified using long‐read sequencing on the PacBio platform. Allele frequencies for HPA systems and HLA class I genes were calculated, and genetic variability within HPA system genes was analysed.ResultsPolymorphisms were identified in 8 of the 35 HPA systems (HPA‐1 to HPA‐6w, HPA‐15 and HPA‐21w), with the frequencies of the ‘b’ allele at 0.0187, 0.0709, 0.4086, 0.0075, 0.0149, 0.0317, 0.4310 and 0.0019, respectively. The alleles with the highest frequencies at the HLA‐A, HLA‐B and HLA‐C loci are HLA‐A02:01, B51:01, B46:01 and C06:02, respectively. Additionally, several genetic patterns in HPA systems were identified, including the c.166‐1029C>T variant, which was found exclusively in samples carrying the HPA‐1b allele.ConclusionThis study developed a targeted long‐read sequencing method characterized by high throughput and simultaneity, capable of resolving allele ambiguities for effective HLA class I genotyping in establishing a platelet donor bank.