【摘要】 目的研究急性髓细胞白血病(AML)患者核仁磷酸蛋白1(NPM1)基因突变检测方法。方法应用PCR结合高分辨熔解曲线分析技术(HRM)和毛细管电泳法(CE)检测正常核型的成人AML样本的NPM1基因突变,并与CE法进行比较分析,结果经克隆测序验证。结果 85例AML样本均得到可供分析的NPM1基因突变检测结果,NPM1基因突变阳性19例,其中15例A型突变,4例B型突变,突变阳性率为22.4%。与CE法比较,二者符合率为100.0%(85/85),HRM检测的敏感性和特异性均达到100.0%。结论应用HRM技术检测NPM1基因突变快速简便,而且敏感,是一项适合临床开展的新方法,可用于临床AML诊断分型、治疗和预后指导。更多还原

【Abstract】 Objective To establish a rapid and simple diagnostic assay for detection of NPM1 mutation of acute myeloid leukemia(AML)using high resolution melting curve analysis(HRM). Methods HRM technology was used to detect NPM1 mutation in 85 adult AML with a normal karyotype samples detected by capillary electrophoresis assay(CE)synchronously,and compared with CE.The positive samples were validated by sequencing. Results The segments of NPM1 gene were successfully amplified,and high-resolution melting curves for all samples were generated,and wild-type samples were definitely and reproducibly discriminated from NPM1-positive samples.NPM1 mutations were present in 22.4%(19/85) of the overall population.Sequence analysis of 19 NPM1 mutated cases showed 15 cases with type-A mutation,4 cases with type-B mutation.The sensitivity and specificity of HRM achieved 100%.When compared to CE,the coincidence rate of NPM1 mutations detected by two assays was 100%(85/85). Conclusion The HRM analysis is a rapid,simple and sensitive method for detection of NPM1 mutation.更多还原