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

EXPRESSION AND ANTIGENICITY TESTING OF RECOMBINANT PROTEINS SJ22.6,LHD-SJ23 AND 2LHD-SJ23 OF SCHISTOSOMA JANPONICUM

作　　者：付媛[1] 石团员[1] 蒲克俊 舒琦艳[1] 卢福庄[1] 帅江冰[3] 张雪娟[1] 莫虹斐 杨富文[1] 何永强 

机构地区：[1]浙江省农业科学院畜牧兽医研究所,杭州310021 [2]兰州工业研究院,兰州730050 [3]浙江出入境检验检疫局检验检疫技术中心,杭州310016

出　　处：《中国动物传染病学报》2017年第2期72-76,共5页Chinese Journal of Animal Infectious Diseases

基　　金：浙江省科技计划项目(2009C12057);国家质量监督检验检疫总局科技项目(2008IK027);浙江出入境检验检疫局重点科技项目(2013-ZKZ-02)

摘　　要：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。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.

关 键 词：日本血吸虫 Sj22.6 LHD-Sj23 2LHD-Sj23 斑点酶联免疫吸附试验 

分 类 号：S852.735[农业科学—基础兽医学]