Preliminary Application Of Recombinant Protein Sj-Tsp6in Immunodiagnosis Of Schistosomiasis Japonica

Objective:In order to discover new candidate diagnosis molecules for schistosomiasis, the SjTsp6protein gene of Schistosoma japonicum were cloned and expressed in the prokaryotic cell. Indirect ELISA (rSj-Tsp6-ELISA) was established for schistosomiasis.Methods:The special primers were designed and synthesized. The target gene was amplified by PCR. The product from PCR was cloned into PMD-T vector, and then was subcloned into the expression vector pET28a (+).After induced by IPTG, the cells were collected to analyze by SDS-PAGE and Western blot. The rSj-Tsp6were expressed in E. coli (BL21) and purified through the column of affinity chromatography with HIS. rSj-Tsp6was coated on sptting plate to determine the optimal amount of antigen and serum dilution, rSj-Tsp6-ELISA were subjected to the serological diagnosis in48samples of rabbit serum infected schistosomiasis japonica.Results:For PCR,, a specific band of around702bp was amplified. The gene was subcloned into the expression vector pET28a (+).After sequencing, the rSj-Tsp6were induced by IPTG and expressed. The results of SDS-PAGE revealed that the molecular weight of fusion protein with HIS was approximately28kDa.The purified rSj-Tsp6was subjected to the serological diagnosis in48samples of rabbit serum infected schistosomiasis japonica and normal rabbit serum using indirect ELISA. Results Display,43samples were positive results, positive rate was89.6%.Conclusion:The cDNA encoding Sj-Tsp6protein of S. japonicum was cloned, expressed in the prokaryotic cell and purified. The rSj-Tsp6antigens were subjected to the serological diagnosis in48samples of rabbit serum infected schistosomiasis japonica using indirect ELISA. The rSj-Tsp6are economic to prepare, easy to standardize, it may be subjected to the serological diagnosis in schistosomiasis in endemic area.