中国动物传染病学报2017，Vol.25Issue(1)：12-18,7.
鸭坦布苏病毒JM株E蛋白的截断表达及间接ELISA方法的建立
ESTABLISHMENT OF AN INDIRECT ELISA ASSAY FOR DETECTION OF ANTIBODIES AGAINST DUCK TEMBUSU VIRUS
摘要
Abstract
The E gene of Duck Tembusu virus,(DTMUV) JM strain was amplified in RT-PCR and cloned into pET32a (+) vector.The recombinant protein was highly expressed in E.coli Rosseta by induction with IPTG and exhibited a favorable reactivity against the DTMUV antibodies in Western blot.The recombinant protein was used as coating antigen to develop an indirect ELISA.The reaction conditions were optimized and determined to be 0.093 μ g/well coating antigen of E protein and 1:100 dilution of antiserum.The intraand inter-assay demonstrated that the coefficient of maximum varation was 3.53％ and 9.73％.Clinical antiserum samples from vaccinated ducks and non-vaccinated ducks were tested using this ELISA.As compared with vaccination-challenge experiment,the results revealed that the correlation rates of positive serum samples from vaccinates and negtive serum samples from non-vaccinated controls were 86.67 ％ and 100 ％,respectively.关键词
鸭坦布苏病毒/JM株/E蛋白/截断表达/ELISAKey words
Duck Tembusu virus/JM strain/E protein/truncated expression/ELISA分类
农业科技引用本文复制引用
董嘉文,李林林,孙敏华,张建峰,邝瑞欢,胡奇林,张春红..鸭坦布苏病毒JM株E蛋白的截断表达及间接ELISA方法的建立[J].中国动物传染病学报,2017,25(1):12-18,7.基金项目
广东省科技计划项目(2013B020307001、2014A040401049、2014A020208052、2015B020203003、2015B070701015、2016A020210049) （2013B020307001、2014A040401049、2014A020208052、2015B020203003、2015B070701015、2016A020210049）
广州市科技计划项目(201510010250) （201510010250）
广东省农业科学院院长基金项目(201436、201412) （201436、201412）
