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目的 利用分子生物学测序技术聚合酶链反应直接测序方法(PCR-SBT))对血小板抗原(HPA)-3系统进行基因分型.方法 采用本研究合成的引物,应用分子生物学测序技术对101名随机血小板捐献者进行HPA-3基因分型,随机抽样20个标本进行聚合酶链反应-序列特异性引物(PCR-SSP)基因分型作为平行对照,质控DNA进行验证.结果 最终选取61℃的为理想退火温度.采用合成的引物对4份质控标本作HPA-3基因分型:与Innotrain提供的结果完全一致.101名随机汉族血小板志愿捐献者HPA基因频率为HPA-3a 0.520、HPA-3b 0.480;随机抽样20个样品与Inno-train公司的HPA-SSP分型结果一致.结论 所建立的HPA基因PCR-SBT分型技术分辨率高,正反向引物测序结果一致,都可以精确分型,可以弥补PCR-SSP不足.“,”Objective Molecular biology sequencing technology was used for genotyping of human platelet antigen-3(HPA-3)system. Methods Primers were designed for the genotyping of HPA-3 system. 101 platelet samples were collected from the donors and the HPA- 3 system genotypes were determined by polymerase chain reaction-sequence based typing(PCR-SBT). 20 samples were genotyped by PCR-SSP for comparison. Results 61℃ was determined as the optimal annealing temperature for PCR-SBT. The results of PCR-SBT and PCR-SSP were consistent. The genotype frequencies of HPA-3a and HPA-3b were 0.520 and 0.480,respectively. Conclusions PCR-SBT assay established in our study provides a high resolution and consistent results. The assay can accurately determine the genotype of HPA- 3 system,and can make up the deficiency of PCR-SSP.