多重PCR结合毛细管电泳检测FLT3与NPM1基因突变的实验方法的研究.pdfVIP

中国血液流变学杂志.2011;21(4) 569 doi:10.3969/j.issn.1009-881X.2011.04.002 多重PCR结合毛细管电泳检测FLT3及NPM1基因突变的实验 方法的研究 薛梦星，仇惠英，冯宇峰，孙爱宁，吴德沛 ＊ （苏州大学附属第一医院血液科，江苏省血液研究所，卫生部血栓与止血重点实验室，江苏苏州 215006） 摘要：目的 建立一种便捷可靠的实验方法能够同时检测FLT3基因突变及NPM1基因突变。方法 设计多重PCR结合毛 细管电泳（CE ）方法同时检测75例初诊急性白血病（AL ）患者FLT3基因及NPM1基因突变发生情况。结果 应用多重PCR 结合CE方法在75例AL患者中检测到FLT3-ITD突变阳性28例，FLT3-TKD突变阳性5例，NPM1突变阳性20例，9例同时存在 FLT3-ITD及NPM1基因突变， 1例同时存在FLT3-ITD及FLT3-TKD突变。结论 多重PCR结合CE是一种能同时检测FLT3基因 和NPM1基因突变的便捷可靠的方法，对判断AML患者的临床预后、指导分层治疗有重要意义，有进一步推广的价值。 关键词：白血病，急性；基因，FLT3 ，NPM1 ；基因突变；多重PCR ；毛细管电泳 中图法分类号：R733.71 文献标识码：A 文章编号：1009-881X(2011)04-0569-04 Simultaneous Detection of NPM1 and FLT3-ITD Mutations by Multiplex PCR Followed with Capillary Electrophoresis XUE Meng-xing,QIU Hui-ying,FENG Yu-feng,et al (Department of Hematology,The First Affi liated Hospital of Soochow University,Jiangsu Institute of Hematology,Key Laboratory of Thrombosis and Hemostasis,Ministry of Health,Suzhou 215006,China) Abstract ：Objective To develop an assay to identify both FLT3 and Nucleophosmin(NPM1) gene mutations in a single multiplex polymerase chain reaction(PCR).Methods Bone marrow mononuclear cells of 75 newly diagnosed acute leukemia(AL) patients were analyzed for both FLT3 and NPM1 mutations by a PCR and capillary electrophoresis(CE) assay.Results Of 75 AL samples analyzed,28 were FLT3-ITD positive,5 were FLT3- TKD positive,20 were NPM1 positive.Among these patients with FLT3 or NPM1 mutations,one case was both FLT3-ITD and FLT3-TKD mutations,nine were both FLT3-ITD and NPM1 mutations.Conclusions Multiplex PCR followed by capillary electrophoresis can be used for a fast and reliable diagnosis of both FLT3 and NPM1 mutations. Key words ：Acute leukemia;