人类血小板抗原-15系统PCR-SBT分型技术的建立

Establishment of genotyping method for human platelet antigens of HPA-15 system by PCR-SBT

目的建立人类血小板抗原(HPA)-15系统的分型方法,并应用于血小板供者库的HPA基因定型。方法采用本研究合成的引物及G&T公司的商品化试剂盒,应用PCR-SBT技术对200名随机的汉族血小板捐献者进行HPA-15系统基因分型,采用第l4届ISBT血小板协作组提供的l0份质控标本以及德国Inno-train公司提供的质控样品作为平行对照,进行验证。结果 200名汉族血小板志愿捐献者HPA基因频率为HPA-15a 0.430、HPA-15b 0.570;基因分型结果与ISBT公布的结果及Inno-train公司提供的质控样品的结果完全一致。结论所建立的HPA基因PCR-SBT分型技术具有高分辨率、高通量、高精确度、能直接发现新的等位基因等特点,具有广泛的应用前景。

Objective To establish a reliable genotyping method for human platelet antigen HPA-15 system by PCR-SBT and to use this method in the further HPA genotyping of volunteer platelet donors.Methods A total of 200 volunteer platelet donors in Guangzhou were genotyped by both our method and the GT American kit at HPA-15system.For quality control,ten coded samples distributed by the l4th Platelet Genotyping and Serology Workshop of the International Society of Blood Transfusion（ISBT） and 4 control samples of Inno-train were genotyped by our method simultaneously.Results A concordance rate of 100% was observed between the results obtained by our established PCR-SBT method and the results provided by ISBT report and Inno-train.The HPA gene frequencies in the 200 random platelet donors were 0.430 and 0.570 for HPA-15a and HPA-15b respectively.Conclusion PCR-SBT assay established in our study provides a high resolution,high throughput and accurate method for HPA-15 system genotyping.The assay can directly discover new allelic genes and shows a broad prospect in its further applications.