摘要目的 探讨高通量流式细胞仪结合序列特异性寡核苷酸探针(FLOW-SSO)技术在人类血小板抗原( HPA)基因分型中的应用.方法 采用FLOW-SSO技术对1378例汉族血小板捐献者HPA1～6,15系统的基因型进行检测；并以10份国际输血协会(ISBT)第12届血小板免疫学术研讨会提供的质控样品对FLOW-SSO技术的可靠性进行考核.以3家供应商提供的不同聚合酶链反应-序列特异引物法(PCR-SSP)试剂,对50份血小板样品的基因分型结果进行比对试验.分型结果采用x2检验考察HPA系统的基因型频率分布是否符合Hardy-Weinberg遗传平衡定律.结果 通过FLOW-SSO方法获得的1379例血小板供者HPA等位基因频率如下,HPA-1a为0.9927,HPA-1b为0.0073,HPA-2a为0.9536,HPA-2b为0.0464,HPA-3a为0.5700,HPA-3b为0.4300,HPA-4a为0.9982,HPA -4b为0.0018,HPA-5a为0.9804,HPA-5b为0.0196,HPA-6a为0.9822,HPA-6b为0.0178,HPA- 15a为0.5337及HPA-15b为0.4663.基因频率检测结果经x2检验,其分布符合Hardy-Weinberg遗传平衡定律.ISBT提供的10份样品考核结果相符率为100％.50份血小板样本采用3种不同的PCR-SSP试剂测定的HPA1～6,15基因分型结果,与FLOW-SSO技术测定结果完全一致.结论 FLOW-SSO基因分型技术有便捷、重复性好及高通量等特点,适用于血小板供者资料库的血小板基因分型.

abstractsObjective To explore the application of the high-throughput flow-sequence specific oligonucleotide probes (FLOW-SSO) technology in human platelet antigen (HPA) genotyping. Method A total of 1378 platelet donors of Han ethnic group were genotyped by FLOW-SSO for HPA system 1-6 and 15 Ten reference samples provided from 12th platelet immunology workshop of International Society of Blood Transfusion (ISBT) were tested as control group, and 50 samples were detected by the PCR-SSP with 3 different manufacturing sources reagents as the comparison test. x2 test was used to study the genotype frequencies of the HPA system, whether conformed the distribution in Hardy-Weinberg genetic equilibrium. Results The distribution of the HPA allelic frequencies of the 1378 donors were as follows:HPA-1a 0.9927 and HPA-1b 0.0073, HPA-2a 0.9536 and HPA-2b 0.0464, HPA-3a 0.5700 and HPA-3b 0.4300, HPA-4a 0.9982 and HPA-4b 0.0018, HPA-5a 0.9804 and HPA-5b 0.0196, HPA-6a 0.9822 and HPA-6b0.0178, HPA-15a 0.5337 and HPA-15b0.4663, which were in agreement with the Hardy-Weinberg genetic equilibrium. Ten reference samples of quality controls were 100％ in concordance with results of ISBT. The 50 genotypes were all matched in comparison of the methods between FLOW-SSO and PCR-SSP. Conclusions With the advantages of rapid, repeatable and high-throughput testing, the streamlined FLOWSSO technology for HPA system 1-6 and 15 genotyping was applicable to large-scare genetic population studies.