Simultaneous genotyping of human platelet alloantigen-1 to 28bw systems by multiplex polymerase chain reaction sequence-based typing
Simultaneous genotyping of human platelet alloantigen-1 to 28bw systems by multiplex polymerase chain reaction sequence-based typing
复制标题
通过多重聚合酶链式反应序列分型对人血小板同种抗原 1 至 28bw 系统进行同步基因分型
DOI：
10.1111/vox.12507
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发表时间：
2017-05-01
期刊：
影响因子：
2.7
通讯作者：
Zhu, F.
中科院分区：
文献类型：
--
作者：
Hong, X.;Chen, S.;Zhu, F.
背景与目的人血小板异体抗原（HPA）基因分型对同种免疫血小板疾病的诊断和预防具有重要意义。本研究采用多重PCR-SBT技术，建立了浙江汉族人群HPA-1 ~ -28bw系统的同步基因分型方法，并对HPA-1 ~ -28bw系统的基因型和等位基因频率进行了分析。材料与方法根据HPA-1 ~ 28bw系统（分别位于ITGB3、GP1BA、ITGA2B、ITGA2、GP1BB和CD109）的核苷酸序列设计特异性引物。采用多重PCR扩增系统，对扩增子进行纯化和测序。共检测到335名健康志愿献血者。结果我院血小板免疫车间10份参考样本的基因型均与已知基因型一致。在28个HPA系统中，中国汉族人群的HPA-1 ~ 6w、HPA-15和HPA-21w系统中检测到HPA a和HPA b等位基因，而在其他HPA系统中仅检测到HPA aa基因型。HPA-1a和HPA-1b的频率分别为0.993和0.007，其中HPA-2a和HPA-2b的频率分别为0.943和0.057，HPA-3a和HPA-3b的频率分别为0.527和0.473，HPA-4a和HPA-4b的频率分别为0.997和0.003，HPA-5a和HPA-5b的频率分别为0.991和0.009，HPA-6wa和HPA-6wb的频率分别为0.980和0.020，HPA-15a和HPA-15b的频率分别为0.508和0.492，HPA-21wa和HPA-21wb的频率分别为0.994和0.006。结论建立了一种HPAs的多重PCR-SBT检测方法，研究数据有助于预防和治疗同种免疫性血小板减少症。
Background and Objectives Human platelet alloantigen (HPA) genotyping is important for the diagnosis and prevention the alloimmune platelet disorders. In this study, a simultaneous genotyping method for HPA-1 to -28bw systems was established using multiplex PCR-SBT and the frequencies of genotypes and alleles of HPA-1 to -28bw systems in the Zhejiang Han population were analysed.Materials and Methods The specific primers were designed according to the nucleotide sequences of HPA-1 to 28bw systems which are located in ITGB3, GP1BA, ITGA2B, ITGA2, GP1BB and CD109, respectively. The multiplex PCR amplification systems were used, and then, the amplicons were purified and sequenced. A total of 335 healthy volunteer blood donors were detected.Results The genotypes of ten reference samples from Platelet Immunology Workshop of ISBT were in concordance with the known genotypes. Among the 28 HPA systems, HPA a and b alleles were found in HPA-1 to 6w, HPA-15 and HPA-21w systems in the Chinese Han population, while only HPA aa genotype was detected in the other HPA systems. The frequencies of HPA-1a and HPA-1b were 0.993 and 0.007, with 0.943 and 0.057 for HPA-2a and HPA-2b, 0.527 and 0.473 for HPA-3a and HPA-3b, 0.997 and 0.003 for HPA-4a and HPA-4b, 0.991 and 0.009 for HPA-5a and HPA-5b, 0.980 and 0.020 for HPA-6wa and HPA-6wb, 0.508 and 0.492 for HPA-15a and HPA-15b and 0.994 and 0.006 for HPA-21wa and HPA-21wb.Conclusions One multiplex PCR-SBT method for HPAs was established and the data of the study could help to prevent and treat for alloimmune thrombocytopenia.