Fusion Expression Of Swine Hepatitis Virus Gene AG02 And Establishment Of Indirect ELISA Diagnosis

Nest RT-PCR was used to amplified swine hepatitis E virus(SHEV) ORF2-AG02 fragment.AG02 was inserted into pET32a vector and the recombinant expression plasmid PET-HEV was constructed. PET-HEV was transferred into BL21(DE3) and induced by 1mM IPTG, the SDS-PAGE result showed that the 33kDa recombinant protein was expressed. The Western blot analysis showed that the recombinant protein has a good antigenicity with positive SHEV serum. An indirected ELISA was established with the purified recombinant protein. An indirected ELISA was established with the purified recombinant protein, after the square matrix titrate determination best density is 2.43 μ g/mL, the blood serum best dilution ratio is 1:40,Compares with ten reagents boxes, proved we establish the ELISA method has the high specificity and the sensitivity.