【摘要】 目的研究慢性骨髓增殖性疾病(CMPDs)获得性JAK2V617F突变检测方法。方法应用高分辨熔解曲线分析技术(HRM),对前期经等位基因特异性PCR法(AS-PCR)分析过的36例CMPDs样本进行JAK2V617F基因突变检测,并与AS-PCR检测情况进行比较分析,结果经测序验证。结果 36例CMPDs样本均得到可供分析的JAK2V617F基因突变HRM检测结果,与AS-PCR法相比较,二者的符合率为97%(35/36);只有一份样本结果不相符,经测序验证,结果与HRM检测相一致;HRM检测的敏感性和特异性均达100%。结论应用HRM技术检测JAK2V617F突变快速简便、而且敏感,是一项适合临床开展的新方法,可用于临床辅助CMPDs的诊断。更多还原

【Abstract】 Aim To establish a rapid and simple diagnostic assay for detection of acquired JAK2V617F mutation of chronic myeloproliferative disorders(CMPDs) using high resolution melting curve analysis(HRM).Methods HRM technology was used to detect JAK2V617F mutation in 36 CMPDs samples detected by allele-specific-PCR assay(AS-PCR) previously,and compared with AS-PCR.The mutations in all samples were validated by direct sequencing.Results The segments of JAK2 gene were successfully amplified,and high-resolution melting curves for all samples were generated,and wild-type samples were definitely and reproducibly discriminated from JAK2V617F-positive samples.The sensitivity and specificity of HRM achieved 100%.When compared to AS-PCR,the coincidence rate of JAK2V617F mutations detected by two assays was 97%(35/36),and only one JAK2V617F-positive sample didn’t match,which actually existed a JAK2V617F mutation verified by sequencing.Conclusion The HRM analysis is a rapid,simple and sensitive method for detection of JAK2V617F mutation.更多还原