日本血吸虫重组蛋白Sj22.6、LHD-Sj23、2LHD-Sj23的表达和抗原性比较

摘要

The membrane-associated proteins Sj22.6 and LHD-Sj23 (large hydrophilic region polypeptide) of Schistosoma japonicum have been shown good antigenicity and used as antigens for the immunological diagnosis of schistosomiasis. In the present study, Sj22.6 and LHD-Sj23 gene fragments were amplified in RT-PCR and 2LHD-Sj23 fragments was amplified in overlapping PCR. Prokaryotic expression plasmids pET28-Sj22.6, pET28-LHD-Sj23 and pET28-2LHD-Sj23 were constructed and expressed in E. coli (Transetta DE3). Three recombinant proteins rSj22.6, rLHD-Sj23 and r2LHD-Sj23 were obtained and tested in Dot-ELISA using positive and negative bovine antisera of schistosomiasis. The results showed that rLHD-Sj23 possessed a better sensitivity and specificity than others. The present study provided preliminary research on recombinant proteins as serological diagnostic antigens for bovine schistosomiasis.%日本血吸虫(Schistosoma japonicum,Sj)膜相关蛋白Sj22.6和Sj23大亲水区多肽(LHD-Sj23)都具有良好的抗原性,可用于日本血吸虫病的免疫学诊断.本研究通过RT-PCR技术扩增了Sj22.6和LHD-Sj23基因片段,重叠延伸PCR方法融合了2个LHD-Sj23的片段,构建了pET28-Sj22.6、pET28-LHD-Sj23以及pET28-2LHD-Sj23原核表达质粒,在大肠杆菌(Transetta DE3)中表达,获得了rSj22.6、rLHD-Sj23和r2LHD-Sj23三种重组蛋白.用Dot-ELISA方法检测牛日本血吸虫阴阳性血清各10份,比较其敏感性和特异性,结果表明rLHD-Sj23具有较好的效果,为进一步研究利用重组蛋白进行血吸虫血清学诊断奠定基础.