Kengkate M, Butthep P, Kupatawintu P, Kanunthong S, Chantratita W, Nathalang O
Department of Pathology, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok, Thailand.
Transfus Med. 2012 Aug;22(4):272-6. doi: 10.1111/j.1365-3148.2012.01153.x. Epub 2012 Apr 10.
Different polymerase chain reaction (PCR) techniques for human platelet antigens (HPA) genotyping have been implemented, in order to diagnose the clinical syndromes of patients with thrombocytopaenia and provide effective HPA-matched platelet donors.
The aim of this study is to develop an in-house multiplex PCR for HPA-1 to -7 and -15 genotyping in the Thai population.
One hundred DNA samples of known HPA genotyping by the PCR with sequence-specific primers (PCR-SSP), as previously described, were tested with the multiplex PCR. Additionally, 300 DNA samples of group O donors were tested for HPA-1 to -7 and -15 genotyping using multiplex PCR.
The comparison of HPA-1 to -7 and -15 genotype results between multiplex PCR and PCR-SSP technique was in 100% concordance. Interestingly, HPA-2b2b genotype was found in two samples; however, other low-incidence genotypes such as HPA-1b1b, HPA-5b5b, HPA-6b6b and HPA-7b7b were not found in this study. Moreover, 30 samples were randomly tested twice for HPA genotyping using the multiplex PCR and demonstrated reproducible results.
This study shows that the in-house multiplex PCR is simple, cost-effective and suitable for HPA genotyping for routine laboratories in other developing countries. Nevertheless, a large-scale evaluation of this technique through multicentre analysis is suggested.
为了诊断血小板减少症患者的临床综合征并提供HPA匹配的血小板供体，已采用不同的聚合酶链反应（PCR）技术进行人类血小板抗原（HPA）基因分型。
本研究旨在开发一种用于泰国人群HPA-1至-7和-15基因分型的内部多重PCR。
用多重PCR检测100份先前用序列特异性引物PCR（PCR-SSP）进行过HPA基因分型的已知DNA样本。此外，用多重PCR对300份O型供体的DNA样本进行HPA-1至-7和-15基因分型检测。
多重PCR与PCR-SSP技术之间HPA-1至-7和-15基因型结果的比较一致性为100%。有趣的是，在两个样本中发现了HPA-2b2b基因型；然而，本研究未发现其他低发生率基因型，如HPA-1b1b、HPA-5b5b、HPA-6b6b和HPA-7b7b。此外，随机选取30个样本用多重PCR进行HPA基因分型检测两次，结果具有可重复性。
本研究表明，内部多重PCR简单、经济有效，适用于其他发展中国家常规实验室的HPA基因分型。尽管如此，建议通过多中心分析对该技术进行大规模评估。
