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#36263028 2022/09/29 To Up
Simultaneous genotyping for human platelet antigen systems and and loci by targeted next-generation sequencing.In order to treat the alloimmunization platelet transfusion refractoriness (PTR), human leukocyte antigen (HLA)-type and/or human platelet antigen (HPA)-type matched platelets between donors and patients are usually used. Therefore, genotyping of and loci, as well as HPA systems, for donors and patients, is of great significance. However, there is a rare report of genotyping for and loci as well as HPA systems at the same time. In this study, a high-throughput method for simultaneous genotyping of and loci, as well as HPA genotyping, was developed. A RNA capture probe panel was designed covering all exon sequences of the , , , , , and genes and and loci. The , , and 34 HPA systems were genotyped using a targeted next-generation sequencing (NGS) method. The genotypes of the and loci, as well as the HPA, were assigned based on the nucleotides in the polymorphism sites. Using the NGS method, 204 unrelated blood specimens were successfully genotyped for all 34 HPA systems as well as and loci. The accuracy of the NGS method was 100%. Only HPA-2, HPA-3, HPA-5, HPA-6w, HPA-15, and HPA-21w showed polymorphism with frequencies of 0.9412, 0.6863, 0.9853, 0.9779, 0.4314, and 0.9951 for a allele, respectively. Thirty-two single nucleotide variants (SNVs) were detected. Of them, 12 SNVs can lead to amino acid change. and are the most common alleles for and loci. A targeted next-generation sequencing method for simultaneously genotyping HPA systems and and loci was first established, which could be used to create a database of HLA-typed and/or HPA-typed unrelated donors.
Jielin Wang, Xuan You, Yanmin He, Xiaozhen Hong, Ji He, Sudan Tao, Faming Zhu