中国畜牧兽医

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犬弓形虫抗体间接ELISA检测方法的建立

贾艳1,陈玉环2,杨丽娜2,刘跃生1   

  1. 1.嘉兴职业技术学院,浙江嘉兴  3140362.北京派克英生物技术有限公司,北京  102200

  • 修回日期:2014-05-13 出版日期:2014-07-20 发布日期:2014-08-21
  • 作者简介:贾艳(1973—),女,四川人,博士,助理研究员,研究方向:兽医微生物与免疫。
  • 基金资助:

    嘉兴市科技局项目(2013AY21035)。

Establishment of Indirect ELISA Detection Method for Toxoplasma gondii Antibody in Dogs

JIA Yan1,CHEN Yu-huan2,YANG Li-na2,LIU Yue-sheng1   

  1. 1. Jiaxing Vocational Technical College,Jiaxing 314036,China;2. Beijing Peking Biological Technical Limited Company,Beijing 102200,China

  • Revised:2014-05-13 Online:2014-07-20 Published:2014-08-21

摘要: 试验以利用重组弓形虫SAG1基因转染蜥蜴利什曼原虫所表达获得的目的蛋白作为包被抗原,建立了一种快速、特异、可检测犬弓形虫抗体的间接ELISA方法。确定了抗原最佳包被浓度为6.75 μg/mL,血清最佳稀释度为1∶100,对已知阳性血清检测的下限可达1∶6400,批间和批内重复性试验的变异系数均小于10%,包被抗原的酶标板在4、-20 ℃环境中可保存8个月以上。建立的间接ELISA方法应用于犬弓形虫抗体的检测具有较好的敏感性及特异性。

关键词: 犬; 弓形虫抗体; 间接ELISA

Abstract: The assay was aimed to establish a rapid and specific indirect ELISA method for canine Toxoplasma gondii antibody detection,using expressed protein of recombinant Toxoplasma gondii SAG1 gene transfected into Leishmania tarentolae as the coating antigen. The optimal antigen coating concentration was 6.75 μg/mL, serum optimal dilution was 1∶100, the known positive serum detection was more than 1∶6400, the coefficients of variation for intra-assay and inter-assay were both less than 10%, and the coated ELISA plate could be stored more than 8 months at 4 and -20 ℃. The present indirect ELISA detection method was sensitivity and specificity for detecting Toxoplasma gondii antibody in canine.

Key words:

canine; Toxoplasma gondii antibody; indirect ELISA