羊血清中口蹄疫病毒非结构蛋白3A-3B1-3B2抗体ELISA检测方法的建立

Establishment of an ELISA method for detecting the nonstructural protein 3A-3B1-3B2 antibody of foot and mouth disease virus in sheep serum

为建立检测羊血清中口蹄疫病毒非结构蛋白抗体的间接ELISA方法,通过优化抗原表达条件,在大肠杆菌原核表达系统中获得了可溶性的3A-3B1-3B2融合蛋白,基于纯化的可溶性融合蛋白建立了口蹄疫病毒非结构蛋白抗体间接ELISA检测试剂盒。该方法对其他相关的羊类病毒病原无交叉反应,其重复性组内与组间变异系数均低于10%,具有良好的重复性。对300份临床血清样品进行检测,与国内市售的口蹄疫非结构蛋白抗体检测试剂盒进行比较,阳性样品符合率为96.67%,阴性样品符合率为94%,总符合率为95.33%。本研究建立的羊血清中口蹄疫病毒3A-3B1-3B2蛋白抗体间接ELISA检测方法,特异性高、敏感性强,操作简单快速,具有较高的应用推广价值。

To establish an indirect ELISA method for detection of foot-and-mouth disease virus antibody in sheep serum, the soluble 3 A-3 B1-3 B2 fusion protein in Escherichia coli was obtained by optimization of protein expression conditions. Further based on purified fusion protein, an indirect ELISA method for detecting the antibody of foot-and-mouth disease virus was established. The results were that this method effectively detected the antibodies of foot-and-mouth disease virus in sheep serum. It had high sensitivity and specificity, but no cross reaction to other related sheep pathogens. The coefficients of variation intra batch and inter batch were both below 10%. A total of 300 clinical serum samples were tested, and the ELISA kit(home-made) was used for comparison with the results of the coincidence rate of the positive samples being 96.67%, the coincidence rate of the negative samples being 94%, and the total coincidence rate being 95.33%. The ELISA method established here was of significant utility in practice and popularization.