The Combined Profile Tests Of Circulating Sj14-3-3 And Its Antibodies And Primary Approach On Proteomic Analysis For The Diagnosis Of Schistosomiasis Japonica

Objective:To evaluate the immunodiagnostic value of the profile tests in detection of both circulating antigen Sj 14-3-3 and its antibodies for the diagnosis of patients with schistosomiasis.Methods:The rSj 14-3-3 expression in E.coli was induced and purified through affinity chromatography.Monoclonal and specific multiclonal antibodies against rSj14-3-3 were produced previously in our laboratory.Sera of patients with acute and chronic schistosomiasis were detected with rSj14-3-3 indirect ELISA,double antibody sandwich ELISA(using anti-rSj 14-3-3 monoclonal and multiclonal antibodies),and the combined detections of Sj 14-3-3 and its antibodies.Results:The positive rates of the three methods in 70 cases of acute schistosomiasis were found to be 72.9%,75.7%,and 91.4%,while in 110 cases of chronic schistosomiasis are 46.4%,61.8%,and 82.7%,respectively.The negative rates in 100 normal persons are 96%,92%,and 92%,while in 18 patients treated with praziquantel are 50%,38.9%,and 33.3%,respectively.The negative rates in 72 chronic cases with one-year post-treatment are 76,4%,55.6%and 43.1%,respectively.The negative rates in 71 chronic subjects with over two-years posttreatment are 93.0%,83.0%and 78.9%, respectively.Cross reactions in sera from 64 cases of clonorchiasis were found to be 0%,15.6%and 15.6%,respectively and from 46 cases of hockworm infection are 8.7%, 19.6%and 21.7%,respectively.Statistically significant difference was noticed in the results,using rSj14-3-3 indirect ELISA,double antibody sandwich ELISA and the combined detection.Conclusion:The profile tests of both circulating antigens and antibodies have a potential value in immunodiagnoses of schistosomiasis. Objective:Most of the antigens involved in the diagnosis of schistosomiasis are the unspecific crude antigens or the single recombinant protein.In our study,we employ the techniques of two-dimensional electrophoresis combined with immunoblotting assay to analyze the components of the crude adult worm antigens of schistosoma japonicum..Methods:First,the crude antigens were separated by two-dimensional electrophoresis, followed by Coomassie staining and immunodetection,respectively,with the use of the sera from normal humans and schistosomiasis patients,which were verified by the egg detection conveniently.Result:About 198 spots were found in Coomassie-stained small gels(7×8 cm,pH ranges 3-10) and imunoblotting of 2-DE showed that there were 39 specific antigen spots with sera from the patients,compared with the controls.The characteristics of of the 39 spots,such as isoelectric point and molecular weight,were analysed with the software from Biorad.We choose 18 to MS detection.Two of those match well.Conclusion:Those should be candidates for diagnostic markers in the immunodiagnosis of schistosomiasis.Then we will continue our study in the analysis of soluble egg antigen and the supematant from culturing the adult worms,which may offer us more information.