AUTHOR=Wang Jielin , You Xuan , He Yanmin , Hong Xiaozhen , He Ji , Tao Sudan , Zhu Faming 

TITLE=Simultaneous genotyping for human platelet antigen systems and HLA-A and HLA-B loci by targeted next-generation sequencing

JOURNAL=Frontiers in Immunology

VOLUME=13

YEAR=2022

URL=https://www.frontiersin.org/journals/immunology/articles/10.3389/fimmu.2022.945994

DOI=10.3389/fimmu.2022.945994

ISSN=1664-3224

ABSTRACT=<p>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 <italic>HLA-A</italic> and <italic>HLA-B</italic> loci, as well as HPA systems, for donors and patients, is of great significance. However, there is a rare report of genotyping for <italic>HLA-A</italic> and <italic>HLA-B</italic> loci as well as HPA systems at the same time. In this study, a high-throughput method for simultaneous genotyping of <italic>HLA-A</italic> and <italic>HLA-B</italic> loci, as well as HPA genotyping, was developed. A RNA capture probe panel was designed covering all exon sequences of the <italic>GP1BA</italic>, <italic>GP1BB</italic>, <italic>ITGA2</italic>, <italic>CD109</italic>, <italic>ITGB3</italic>, and <italic>ITGA2B</italic> genes and <italic>HLA-A</italic> and <italic>HLA-B</italic> loci. The <italic>HLA-A</italic>, <italic>HLA-B</italic>, and 34 HPA systems were genotyped using a targeted next-generation sequencing (NGS) method. The genotypes of the <italic>HLA-A</italic> and <italic>HLA-B</italic> 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 <italic>HLA-A</italic> and <italic>HLA-B</italic> 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. <italic>HLA-A*11:01</italic> and <italic>HLA-B*46:01</italic> are the most common alleles for <italic>HLA-A</italic> and <italic>HLA-B</italic> loci. A targeted next-generation sequencing method for simultaneously genotyping HPA systems and <italic>HLA-A</italic> and <italic>HLA-B</italic> loci was first established, which could be used to create a database of HLA-typed and/or HPA-typed unrelated donors.</p>