口蹄疫病毒非结构蛋白定量检测ELISA方法的建立  

The Establishment of Quantitative ELISA of Foot-and-Mouth Disease Virus Non-structural Protein 3B

作　　者：李永亮[1] 卢曾军[1] 杨苏珍[2] 田美娜[1] 谢宝霞[1] 刘在新[1] 

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室农业部畜禽病毒学重点实验室国家口蹄疫参考实验室,兰州730046 [2]河南省农业科学院动物免疫学重点实验室,郑州450002

出　　处：《中国生物工程杂志》2009年第11期70-73,共4页China Biotechnology

基　　金：国家"973"项目(2005CB523201)资助项目

摘　　要：目的:利用口蹄疫病毒非结构蛋白3B单克隆抗体建立液相阻断ELISA检测方法,进行定量检测口蹄疫病毒培养液中的非结构蛋白含量。方法:首先将工作浓度的3B单抗与待测病毒培养液过夜结合反应,然后取结合液转移至用3B蛋白包被好的酶标板上,用标准3B蛋白做12个梯度做对照,同时设阴性对照和空白对照。通过回归分析算出口蹄疫病毒培养液中的非结构蛋白3B含量。结果:回归曲线呈典型的S形,符合4参数logit曲线拟合,相关系数R=0.99,检测范围为5～1500ng/ml,半数抑制浓度(Ic50)为130ng/ml。结论:该方法能特异、敏感的检测到病毒培养液中的非结构蛋白3B成分,并进行定量。Objective：Establish an ELISA method for quantitative detection of non-structural protein 3B in the FMD virus culture medium using monoclonal antibody. Methods：At first, monoclonal antibody was binding with the test virus culture medium overnight, and then transferred to the ELISA plate coated with 3B protein, using standard protein 3B in control, while establishment negative and blank control. The content of nonstructural protein 3B in the culture medium was calculated by regression analysis finally. Results：The regression curve was typical of s-type and fitted with the 4 parameter logic curves. The correlation coefficient R was 0.99 and the range of detection was 5 - 1500ng/ml while the half inhibitory concentration （Ic50） was 130ng/ml. Conclusion：This method is specific, sensitive and quantitative to detect non-structural protein 3B in the virus culture medium.

关 键 词：口蹄疫病毒 单克隆抗体 非结构蛋白 定量ELISA 

分 类 号：Q511[生物学—生物化学]