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以重组蛋白为抗原的ELISA法检测旋毛虫抗体
引用本文:牛廷献,潘菲菲,刘明远,卢强,付宝权,Pascal Boireau.以重组蛋白为抗原的ELISA法检测旋毛虫抗体[J].中国寄生虫学与寄生虫病杂志,2005,23(3):143-145.
作者姓名:牛廷献  潘菲菲  刘明远  卢强  付宝权  Pascal Boireau
作者单位:1. 兰州军区总医院动物实验科,兰州,730050
2. 兰州军区五里铺干休所,兰州,730050
3. 解放军军需大学军事兽医系,长春,130062
4. 法国食品卫生安全局,法国,巴黎,94703
基金项目:国家自然科学基金(NSFC-30170709),中法先进计划项目(9612)~~
摘    要:目的 寻求特异性强、敏感性高的旋毛虫病诊断抗原。 方法 以旋毛虫新生幼虫期特异性T668基因在E.coli高效表达的重组蛋白为抗原,分别以兔、猪和健康者血清及旋毛虫病阳性血清为一抗,以辣根过氧化物酶(HRP)标记的山羊抗兔IgG、山羊抗猪IgG和山羊抗人IgG为二抗,建立检测旋毛虫抗体的间接ELISA方法,并以旋毛虫肌幼虫排泄?鄄分泌(ES)抗原作为检测对照。 结果 以T668重组蛋白为抗原,对兔、猪和人旋毛虫病血清进行检测,阳性检出率为100 %,且敏感性高(0.016 μg / 孔),与ES抗原检测结果完全一致。 结论 T668重组抗原有望替代ES抗原检测旋毛虫抗体。

关 键 词:旋毛虫  重组蛋白  新生幼虫期特异性基因  间接ELISA
文章编号:10000-7423(2005)-03-0143-03
修稿时间:2004年9月16日

Detection of Anti-Trichinella spiralis Antibody by Indirect ELISA Using Recombinant Protein as Antigen
NIU Ting-xian,PAN Fei-Fei,LIU Ming-yuan,LU Qiang,FU BAO-QUAN,Pascal Boireau.Detection of Anti-Trichinella spiralis Antibody by Indirect ELISA Using Recombinant Protein as Antigen[J].Chinese Journal of Parasitology and Parasitic Diseases,2005,23(3):143-145.
Authors:NIU Ting-xian  PAN Fei-Fei  LIU Ming-yuan  LU Qiang  FU BAO-QUAN  Pascal Boireau
Institution:Lanzhou General Hospital of PLA, Lanzhou 730050, China.
Abstract:OBJECTIVE: To study the specificity, sensitivity of diagnostic antigen for Trichinella spiralis (T.s). METHODS: T668 recombinant protein from highly efficient expression of newborn larvae stage-specific gene of T.s in E. coli as diagnostic antigen. By using negative and positive sera from rabbit, pig, human as the first antibody, and goat-anti-rabbit IgG, goat-anti-pig IgG, goat-anti-human IgG labeled with HRP as the secondary antibody, indirect ELISA method was established for detecting anti-T.s antibody, while excretory-secretory (ES) antigen of T.s muscle larvae was used as control. RESULTS: Sera from rabbit, pig and human were detected by T668 recombinant protein as antigen, which showed a positive rate of 100% with 0.016 microg/well. There was no difference on the results between the T668 recombinant antigen and the ES antigen for diagnosing T.s-infection. CONCLUSION: The T668 recombinant antigen is promising in substituting ES antigen in the detection of anti-T.s antibody.
Keywords:Trichinella spiralis  Recombinant protein  Newborn larvae (NBL) stage-specific gene  Indirect ELISA
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