牛日本血吸虫重组二价表位抗原间接ELISA检测方法的建立及应用  

Establishment and application of ELISA for diagnosis of bovine schistosomiasis using Schistosoma japonicum bivalent epitope antigen

作　　者：陆珂[1] 李浩[1] 石耀军[1] 周伟芳[1] 陈实[1] 朱传刚[1] 傅志强[1] 杨健美[1] 洪炀[1] 苑纯秀[1] 林矫矫[1] 

机构地区：[1]中国农业科学院上海兽医研究所国家防治动物血吸虫病专业实验室农业部动物寄生虫病重点开放实验室,上海200241

出　　处：《中国兽医科学》2012年第12期1273-1277,共5页Chinese Veterinary Science

基　　金：公益性行业(农业)科研专项(200903036)

摘　　要：为了建立一种更为敏感、特异的牛血吸虫病诊断方法,表达和纯化了日本血吸虫二价表位抗原(pGEX-BSjGCP-BSj23),将重组蛋白以10mg/L浓度包被酶标板,以1∶40 000稀释酶标兔抗牛IgG,建立了检测牛血吸虫病的间接ELISA方法。以SEA抗原作为对照,进行敏感性试验和特异性试验。结果显示,以重组抗原pGEX-BSjGCP-BSj23作为诊断抗原,建立的间接ELISA的特异性明显高于SEA,敏感性与SEA相当。检测现场水牛血清样品,结果阴性符合率为100%,阳性符合率为90.97%。结果表明,建立的ELISA检测方法对牛血吸虫病的诊断具有良好的特异性和敏感性。In order to establish a sensitive and specific method for diagnosis of schistosomiasis,the expression and purification of Schistosoma japonicum bivalent epitope antigen（pGEX-BSjGCP-BSj23） was carried out.Using the recombinant protein（10 mg/L） and 1∶40 000 diluted HRP rabbit anti-bovine IgG,an indirect ELISA was developed for diagnosis of bovine schistosomiasis.SEA antigen was used as a control to compare the sensitivity and specificity of the developed ELISA.In result,the bivalent epitope antigen based ELISA was significantly more specific than the method based on SEA,and was similar in sensitivity.Regarding the buffalo sera detected by the developed ELISA,the negative coincidence rate was 100% and positive coincidence rate was 90.97%.The result showed that the developed ELISA was specific and sensitive for diagnosis of bovine schistosomiasis.

关 键 词：日本血吸虫 二价表位抗原 间接酶联免疫吸附试验 

分 类 号：S852.735[农业科学—基础兽医学]