猪嵴病毒重组VP3蛋白间接ELISA检测方法的建立

Establishment of an indirect ELISA based on VP3 protein for the detection of the antibodies against porcine kobuvirus

为建立检测猪嵴病毒(porcine kobuvirus,PKV)血清抗体的间接ELISA检测方法,以原核表达的重组VP3蛋白为检测抗原,优化ELISA检测条件,建立了PKV抗体的间接ELISA检测方法。该方法仅对PKV血清检测为阳性,对于能够引起猪病毒性腹泻的猪流行性腹泻病毒、猪传染性胃肠炎病毒以及猪轮状病毒阳性血清检测均为阴性。临床血清样品的检测结果表明,建立的ELISA检测方法与RT-PCR方法的阳性符合率较高。结果表明,建立的以重组VP3蛋白为包被抗原检测PKV血清抗体的ELISA方法可以用于检测PKV的感染及相关流行病学调查。

To establish a serological diagnosis method of porcine kobuvirus （PKV） infection,an indirect ELISA was developed for the detection of PKV antibodies with the VP3 protein as a coating antigen expressed in Escherichia coli.Under the optimized reaction conditions,the assay was able to specifically detect the positive sera against PKV,without cross-reactions with porcine epidemic diarrhea virus,transmissible gastroenteritis virus of swine,or porcine rotavirus,which can cause diarrhea in pigs too.Detection results showed that RT-PCR and ELISA were well consistent,which manifested the ELISA was a useful method to investigate the prevalence of PKV.