非洲猪瘟病毒p54抗体胶体金试纸检测方法的建立

Establishment of colloidal gold strip for detecting antibody against African swine fever virus

为建立非洲猪瘟病毒（ASFV）p54抗体的胶体金检测方法，本研究原核表达并纯化了带有His标签的p54重组蛋白（aa51～aa183），胶体金标记后，喷涂玻璃纤维垫，分别以SPA和抗p54多克隆抗体作为检测线和质控线，制作ASFVp54抗体检测胶体金免疫层析试纸。检测结果表明，该试纸条的敏感性为200ng／mL，对ASFV抗体阳性猪血清具有高度特异性，与猪其他病毒抗体阳性血清无交叉反应。对141份猪血清样品进行比对检测表明，在ASFV感染早期，胶体金试纸优于OIE推荐间接ELISA检测方法；以western blot检测方法作为参照，该试纸条检测疫区临床阳性样品的符合率比OIEELISA高。表明本研究建立的ASFV抗体免疫层析检测方法具有特异性强、敏感性高等特点，在ASF防控中具有趣好的应用前景。

To establish a method of colloidal gold strip for the detection of antibodies against African swine fever virus （ASFV）, the purified recombinant p54 （51aa-183aa） of ASFV expressed in E.coli was labeled with colloidal gold as a detection reagent, SPA and polyclonal antibodies to p54 were blotted on nitrocellulose membrane as test line and control line, respectively. The test results showed that the strips were specific for ASFV positive serum, and no cross reactions for positive sera to CSFV, PRV, PRRSV, PPV and PCV-2. Comparing with OIE recommended indirect ELISA and western blot, the strip was more efficient than that of the indirect ELISA in early antibody detection, and had a higher positive coincidence rate to western blot in detecting clinical samples of 141 porcine sera, which indicated that the colloidal gold strip was highly specific and sensitive, and had a potential application in ASF detection.