猪流行性腹泻病毒S1蛋白B细胞表位原核表达及间接ELISA方法的建立

Prokaryotic expression of one B cell epitope in S1 protein of porcine epidemic diarrhea virus and establishment of indirect ELISA

应用DNAStar软件在猪流行性腹泻病毒S1蛋白70~280位氨基酸序列间预测出1个优势B细胞抗原表位,氨基酸序列为IARLRICQFP,命名为PEP3。借助基因工程技术成功对PEP3进行融合表达,经动物试验、Western blot和间接免疫荧光试验均表明,PEP3蛋白具有良好的抗原性。基于PEP3蛋白建立的ELISA检测方法,与用N蛋白建立的检测方法的符合率为95.94%;该方法具有较高的敏感性和良好的特异性,批内和批间变异系数分别为1.68%~4.65%和4.21%~9.32%。研究表明,本试验建立的基于PEP3蛋白的ELISA检测方法特异性强、敏感性好、重复性高,可以用于临床样本的检测,丰富了现有的猪流行性腹泻病的诊断防控技术。

A dominant B cell epitope on the porcine epidemic diarrhea virus(PEDV)was predicted by using DNAStar software in the 70-280 amino acid sequence of S1 protein,the amino acid sequence was IARLRICQFP,and named PEP3.The fusion expression of PEP3 was successfully carried out by gene engineering technology.The results of animal experiment,Western blot and indirect immunofluorescence showed that PEP3 protein had good antigenicity.Based on the PEP3 protein,an indirect ELISA method was established,which was compared with the ELISA method established by N protein,the coincidence rate with the established ELISA method were 95.94%.This method had high sensitivity and excellent specificity,respectively.Repeatability tests showed that the coefficient of variation of serum samples within and among runs were 1.68%-4.65% and 4.21%-9.32%,respectively.The results suggest that this method is simple to perform,and is a specific,sensitive and convenient method.The method can be used for the detection of clinical samples,and enrich the existing porcine epidemic diarrhea prevention and control technology.