口蹄疫病毒非结构蛋白定量检测ELISA方法的建立

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。

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

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 non-structural 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.