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Cloning, molecular characterization of a 13-kDa antigen from Schistosoma japonicum, Sj13, a putative salivary diagnosis candidate for Schistosomiasis japonica

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Abstract

Saliva has been suggested as an easily accessible and a noninvasive diagnostic alternative for detection of antibodies. To identify and characterize Schistosoma japonicum (S. japonicum) antigens that are recognized by saliva of infected host, we have used a pool of saliva from infected patients to immunoscreen an egg cDNA library of S. japonicum. The open reading frame of the isolated two clones encodes same protein of 116 amino acids exhibiting 100% identity to an amino acid sequence (AY222893) of S. japonicum in NCBInr database. The protein encoded is inferred a secretory protein with a molecular mass of 13 kDa (Sj13) and shares no homology to any entries in the NCBInr database, demonstrating that Sj13 might be a schistosome-specific protein. Reverse transcriptase polymerase chain reaction, Western blotting, and immunolocalization analysis revealed Sj13 could be detected in cercaria, adult, and egg and was localized to forehead and tegument of cercaria, cell body (“cytons”) of adult worm, egg shell, and epidermal plate of miracidium. Furthermore, Sj13 showed a good antigenicity when reacted with saliva or serum from schistosomiasis patients. The recombinant Sj13 (rSj13) expressed and purified from Escherichia coli was applied to detect its specific salivary antibody for schistosomiasis diagnosis by an indirect enzyme linked immunosorbent assay (ELISA). Preliminary laboratory test of 116 subjects, 40 with parasitologically proven S. japonicumm infection, 46 with other infectious diseases, and 30 negative controls exhibited 92.50% sensitivity with saliva/rSj13 and 95.00% with serum/SWAP (P > 0.05). The specificity of the ELISA using saliva/rSj13 was 92.11% versus 85.53% with serum/SWAP (P < 0.05). No direct correlations of anti-Sj13 IgG levels with egg counts in stool were observed in saliva detection. These results suggest that Sj13 specific salivary antibody detection may be useful as an antigen for the salivary diagnosis of schistosomiasis japonica and contribute to epidemiological study of schistosomiasis infection in endemic areas.

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Acknowledgements

We thank Prof. Wu Haiwei for a gift of S. japonicum egg cDNA library, Prof. Chen Jiaxu and Mr. Lin An for kindly presenting saliva and sera samples, and Prof. Fung Ming-Chiu for helping with mass spectrometry and expert technical assistance. This work was financed by National Natural Science Foundation of China (grant nos. 30771888, 30800966), National High-Tech Research and Development Program of China (863 Program; grant nos. 2006AA02Z444, 2007AA02Z153), and Project of Basic Platform for National Science and Technology Resources of the Ministry of Sciences and Technology of China (grant no. 2005DKA21104).

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  1. Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, 74 Zhongshan 2nd Road, Guangzhou, 510080, China

    Yan-ping Zhou, Zhong-dao Wu, Lin-lin Yang, Xi Sun, Xu You, Xin-bing Yu, Huan-qin Zheng & Zhi-yue Lv

  2. Key Laboratory for Tropical Diseases Control, Ministry of Education, Sun Yat-sen University, Guangzhou, 510080, China

    Yan-ping Zhou, Zhong-dao Wu, Lin-lin Yang, Xi Sun, Xu You, Xin-bing Yu, Huan-qin Zheng & Zhi-yue Lv

  3. Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, 207 Rui-Jin Road II, Shanghai, 200025, China

    Wei Hu

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  1. Yan-ping Zhou
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Correspondence to Zhi-yue Lv.

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Zhou, Yp., Wu, Zd., Yang, Ll. et al. Cloning, molecular characterization of a 13-kDa antigen from Schistosoma japonicum, Sj13, a putative salivary diagnosis candidate for Schistosomiasis japonica . Parasitol Res 105, 1435–1444 (2009). https://doi.org/10.1007/s00436-009-1575-x

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