大豆花叶病毒间接ELISA检测方法的建立及应用  

Establishment and Application of Indirect ELISA of Mosaic Virus of Soybean

作　　者：李小宇[1] 张春雨[1] 张伟[1] 苏前富[1] 苏颖[1] 张金花[1] 李建平[1] 王永志[1] LI Xiaoyu;ZHANG Chunyu;ZHANG Wei;SU Qianfu;SU Ying;ZHANG Jinhua;LI Jianping;WANG Yongzhi(Jilin Academy of Agricultural Sciences/Key Laboratory of Integrated Pest Management on Crops in Northeast, Ministry of Agriculture /Jilin Key Laboratory of Agricultural Microbiology,Gongzhuling 136100, China)

机构地区：[1]吉林省农业科学院/东北作物有害生物综合治理重点实验室/吉林省农业微生物重点实验室,吉林公主岭136100

出　　处：《东北农业科学》2019年第1期22-27,共6页Journal of Northeast Agricultural Sciences

基　　金：吉林省农业科技创新工程自由创新项目(CXGC2017ZY035);农业部东北作物有害生物综合治理重点实验室开放基金课题(DB201505KF08)

摘　　要：为快速有效地检测大豆花叶病毒(Soybean Mosaic Virus,SMV),本研究利用已制备的大豆花叶病毒衣壳蛋白(Coat Protein,CP)及其单克隆抗体,通过优化ELISA条件为:检测材料抗原包被酶标板37℃1 h,检测抗体工作浓度125 ng/mL,孵育60 min,酶标抗体工作浓度100 ng/mL,孵育30 min;SMV CP蛋白最低检测限为3.23 ng/mL;该方法重复性变异系数小于3%,重复性良好;运用上述检测条件与RTPCR检测方法对田间样品进行随机检测,结果显示一致率高达94%。SMV间接ELISA检测方法的成功建立,为SMV快速检测试剂盒及试纸条的研发奠定了基础。In order to detect mosaic virus of soybean rapidly and effectively, we established indirect ELISA detection method by using prepared monoclonal antibody of soybean mosaic virus and optimizing the influence factors.The antigen was coated onto the microtiter plates at 37℃ for 1 h. The detection antibody at 125 ng/mL was incubated for 1 h. The enzyme labeled antibody at 100 ng/mL was incubated for 30 min. The detection sensitivity was 3.23 ng/mL for smv cp protein. The coefficient of variation of reproducibility was less than 3%. when the developed indirect ELSIA method was compared with RT-PCR, the concordance rate was 94%. Establishment of indirect ELISA for soybean mosaic virus coat protein laid the foundation for development of detection kit and test strip.

关 键 词：大豆花叶病毒 间接ELISA 检测 

分 类 号：S435.651[农业科学—农业昆虫与害虫防治]