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首页> 外文期刊>Transfusion: The Journal of the American Association of Blood Banks >Mass-scale high-throughput multiplex polymerase chain reaction for human platelet antigen single-nucleotide polymorphisms screening of apheresis platelet donors.
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Mass-scale high-throughput multiplex polymerase chain reaction for human platelet antigen single-nucleotide polymorphisms screening of apheresis platelet donors.

机译：大规模高通量多重聚合酶链反应用于单采血小板供体的人血小板抗原单核苷酸多态性筛选。

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摘要

BACKGROUND: Treatment with human platelet antigen (HPA)-matched platelets (PLTs) is the optimal therapy for bleeding secondary to neonatal alloimmune thrombocytopenia. Recent advances in high-throughput DNA-based blood group and PLT antigen genotyping have made it possible to screen plateletpheresis donors for potential HPA-matched PLT transfusion. STUDY DESIGN AND METHODS: This prospective study evaluated genomic DNA from plateletpheresis donors for single-nucleotide polymorphisms (SNPs) associated with HPA-1, -2, -3, -4, -5, and -15 to determine whether high-throughput multiplex genomic DNA PCR and oligonucleotide extension technology can be used for mass-scale PLT antigen genotyping. Genotyping using SNP technology was confirmed using sequence-specific polymerase chain reaction (SSP-PCR). RESULTS: Of the 748 donors screened, 277 were found to be negative for antigens implicated in alloimmune thrombocytopenia. In addition, two donors were homozygous for HPA-1b/b and -2b/b, six donors for HPA-1b/b and -3b/b, one for HPA-2b/b and -3b/b, one for HPA-1b/b and -5b/b, 10 for HPA-1b/b and -15 b/b, four for HPA-5b/b and -15b/b, and one for HPA-2b/b and -15b/b. Retesting using SSP-PCR was conducted for 60 donors. Discrepant results occurred between SNP and SSP-PCR in less than 20% of samples for HPA-1b/1b/HPA-3b/3b, HPA-5b/5b, and HPA-15b/b. DISCUSSION: High-throughput multiplex PCR SNP and confirmatory molecular genotyping are useful for mass-scale screening of apheresis PLT donors to provide antigen-negative genotypes. Refinements to mass-scale multiplex analysis technology would reduce further the confirmatory testing needed.
机译：背景：人类血小板抗原（HPA）匹配血小板（PLTs）的治疗是新生儿同种免疫血小板减少症继发性出血的最佳治疗方法。高通量基于DNA的血型和PLT抗原基因分型的最新进展使得有可能筛选血小板减少供体以寻找潜在的HPA匹配的PLT输血。研究设计和方法：这项前瞻性研究评估了来自血小板减少供体的基因组DNA与HPA-1，-2，-3，-4，-5和-15相关的单核苷酸多态性（SNP），以确定是否高通量多重基因组DNA PCR和寡核苷酸延伸技术可用于大规模PLT抗原基因分型。使用序列特异性聚合酶链反应（SSP-PCR）证实了使用SNP技术进行基因分型。结果：在筛选的748名供体中，发现277例与同种免疫血小板减少症有关的抗原阴性。此外，有两个供体是HPA-1b / b和-2b / b纯合子，六个供体是HPA-1b / b和-3b / b，一个是HPA-2b / b和-3b / b，一个是HPA- 1b / b和-5b / b，HPA-1b / b和-15b / b为10，HPA-5b / b和-15b / b为4，HPA-2b / b和-15b / b为1。使用SSP-PCR对60个供体进行了重新测试。对于HPA-1b / 1b / HPA-3b / 3b，HPA-5b / 5b和HPA-15b / b，少于20％的样品在SNP和SSP-PCR之间产生了不一致的结果。讨论：高通量多重PCR SNP和确定性分子基因分型可用于大规模筛选单采血液分离术PLT供体，以提供抗原阴性基因型。对大规模多重分析技术的改进将进一步减少所需的验证测试。

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