2010•Chinese Veterinary ScienceRequires access
Establishment of iELISA based on the expressed bovine rotavirus VP6 protein for detection of specific antibody in serum
Yiting Zhang, Wang JieQing, Guanqun Zhang, Junwei Ge, Xinyuan Qiao, Yijing Li
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Abstract
The VP6gene was amplified from bovine rotavirus(RV)by RT-PCR and sub-cloned into the prokaryotic expression vector pProHTa,then the construction of pProHTa-NCDV-VP6was finished,and the expression of the recombinant protein was carried out under induction of IPTG.SDS-PAGE and Western-blot analyses showed that the recombinant protein was 52ku approximately in size.The expressed protein VP6was used as coating antigen,and an indirect ELISA was developed to detect antibody against bovine RV in serum.The detection system was 1×104 times sensitive than the traditional agar diffusion test, indicating that the developed indirect ELISA was a simple,rapid,sensitive and specific method for the detection of the antibody against bovine RV in practice.