Abstract
BackgroundSchistosomiasis is a chronic disease caused by trematode flatworms of the genus Schistosoma. The disease remains a serious public health problem in endemic countries and affects at least 207 million people worldwide. A definite diagnosis of the disease plays a key role in the control of schistosomiasis. The detection of schistosome circulating antigens (CAs) is an effective approach to discriminate between previous exposure and current infection. Different methods have been investigated for detecting the CAs. However, the components of the schistosome CAs remain unclear. In this study, we analyzed the CAs in sera of patients infected with Schistosoma japonicum.MethodsThe parasites were collected from the infected rabbits for preparing the adult worm antigen (AWA). The hyline hens were immunized subcutaneously with AWA to produce anti-AWA IgY. The IgY was purified by water-dilution and ammonium sulfate precipitation method and identified by ELISA and Western blotting. After purification and characterization, IgY was immobilized onto the resin as a capture antibody. The circulating antigens were immune-precipitated from patients′ serum samples by direct immunoprecipitation. The precipitated proteins were separated by one-dimensional electrophoresis and analyzed by LC-MS/MS.ResultsFirstly, the IgY against AWA was produced from the eggs of immunized hens by AWA, which gave a titer of 1:12800. The purified IgY was used as the capture antibody to enrich the CAs in sera of S. japonicum infected patients through immunoprecipitation. The CAs were determined by LC-MS/MS. There were four proteins, including protein BUD31 homolog, ribonuclease, SJCHGC06971 protein and SJCHGC04754 protein, which were identified among the CAs.ConclusionsWe developed a novel method based on IgY for identification and profiling CAs in sera of S. japonicum infected patients. Four new CAs were identified and have potential value for further development of an antigen assay.
Highlights
Schistosomiasis is a chronic disease caused by trematode flatworms of the genus Schistosoma
Analysis of IgY We used the water dilution method to obtain a supernatant with crude egg yolk antibody, and the IgY was precipitated by ammonium sulfate with better purity
The results indicated that cathodic antigen (CCA) is O-glycosylated mostly via GalNAc-Thr and circulating anodic antigen (CAA) is a glycoprotein, O-glycosylated at Thr
Summary
Schistosomiasis is a chronic disease caused by trematode flatworms of the genus Schistosoma. Schistosomiasis, known as Bilharziasis, is a parasitic disease caused by trematode flatworms of the genus Schistosoma. Schistosomiasis can be diagnosed by direct or indirect methods: a) direct parasitological examinations to detect parasite eggs in fecal/ urine samples or in the tissues; b) direct approaches to detect the schistosome-derived antigens in the circulation and excreta; c) indirect immunological tests to detect the specific antibodies induced against the different stages of the parasite in blood [7,8]. Direct parasitological diagnosis techniques are labor-intensive and time-consuming Their low sensitivities would result in under-estimation in prevalence and infection intensity, in the areas with low prevalence or after intervention [9,10]. Antibody-based serological assays do not discriminate between active and past infections, and could not be used to evaluate therapeutic efficacy since specific antibodies continue to be present for a long time after the worms have disappeared [10,11]
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