应用荧光实时定量PCR方法检测重组慢病毒滴度及其感染效率  

A Novel Method for the Determination of Recombinant Lentiviral Titer and Infectivity by qRT-PCR

作　　者：马海燕[1] 方彧聃[1] 张敬之[1] 

机构地区：[1]上海交通大学医学院上海市儿童医院上海市医学遗传研究所,中国上海200040

出　　处：《生命科学研究》2009年第5期394-398,共5页Life Science Research

基　　金：国家高技术研究发展计划项目(2007AA021206);国家自然科学基金资助项目(30870943);上海市自然科学基金资助项目(08ZR1412100)

摘　　要：慢病毒载体已经广泛应用于动物模型中基因治疗的研究和转基因动物的制备,而准确地测定重组慢病毒的滴度和感染效率是其关键步骤.通过荧光实时定量PCR的方法定量分析重组慢病毒的颗粒数以及病毒的活性滴度,并以GFP报告基因的方法作为对照来验证定量PCR方法的准确性.研究结果显示,应用荧光实时定量PCR法与GFP报告基因法测定得到的病毒活性滴度成正相关,而且前者可以更加准确地测定病毒滴度和病毒感染效率.Lentiviral vector is being widely used in the study of gene therapy in animal models and in generating transgenic animals. However, determination of lentiviral particles and their infectivity is essential before their being used. Such a requirement can be accurately achieved by qRT-PCR. Refered by infectious units got from GFP reporter assay, it showed a positive correlation between the two approaches. A reliable, accurate and rapid method is therefore established for the determination of the recombinant lentiviral titer and the infectivity.

关 键 词：慢病毒载体 荧光实时定量PCR 病毒滴度 整合拷贝数 感染效率 

分 类 号：Q331[生物学—遗传学]