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| 传染性支气管炎病毒抗体间接ELISA检测方法的建立 | |
| 引用本文： | 高峰,唐应华,宫玉珍,陈涛,陆吉虎,吴培培,傅童生,侯继波.传染性支气管炎病毒抗体间接ELISA检测方法的建立[J].中国家禽,2012,34(8):28-31. |
| 作者姓名： | 高峰 唐应华 宫玉珍 陈涛 陆吉虎 吴培培 傅童生 侯继波 |
| 作者单位： | 1. 湖南农业大学,湖南长沙410128;江苏省农业科学院国家兽用生物制品工程技术研究中心,江苏南京210014 2. 江苏省农业科学院国家兽用生物制品工程技术研究中心,江苏南京,210014 3. 湖南农业大学,湖南长沙,410128 |
| 基金项目： | 江苏省农业科技自主创新项目 |
| 摘 要： | 为了快速检测针对传染性支气管炎病毒(IBV)抗体,本研究建立相应的间接ELISA(iELISA)检测方法。以本实验室分离的临床毒株DS10为检测抗原,经过差速离心纯化获得DS10病毒作为包被抗原,利用方阵试验,优化一系列条件,最终确定了最佳的ELISA反应条件。抗原最佳包被浓度为0.11mg/mL,血清最适稀释度为1:100,封闭液为含1%BSA的PBST,封闭时间为60min,一抗作用时间90min,HRP标记的兔抗鸡IgG稀释倍数为1:3500,作用时间为60min,显色时间为10min。用iELISA方法和血凝抑制试验(HI)同时检测145份血清样品的结果表明,建立的iELISA方法与常规的HI检测的符合率为92.41%,iELISA和HI的阳性检出率分别为92.41%和90.34%,iELISA的敏感性略优于HI。 |
| 关 键 词： | 传染性支气管炎 间接ELISA 抗体检测 |
Development of Indirect ELISA for Detection of Antibodies Against Infectious Bronchitis Virus |
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| GAO Feng , TANG Yinghua , GONG Yuzhen , CHEN Tao , LU Jihu , WU Peipei , FU Tongsheng , HOU Jibo.Development of Indirect ELISA for Detection of Antibodies Against Infectious Bronchitis Virus[J].China Poultry,2012,34(8):28-31. | |
| Authors: | GAO Feng TANG Yinghua GONG Yuzhen CHEN Tao LU Jihu WU Peipei FU Tongsheng HOU Jibo |
| Institution: | 1.College of Veterinary Medicine,Hunan Agricultural University,Changsha,Hunan 410128;2.National Veterinary Biological Medicine Engineering Research Center,Jiangsu Academy of Agricultural Sciences,Nanjing,Jiangsu 210014) |
| Abstract: | The aim of this study was to develop an indirect ELISA(iELISA) method to detect antibody against infectious bronchitis virus.Isolate DS10 was used as coating antigen which was purified by differential centrifugation.The iELISA parameters were optimized by checkerboard titration experiments.The optimum parameters included coating antigen with concentration of 0.11 mg/mL,tested serum with dilution of 1:100,1%BSA-PBST as blocking buffer with blocking time of 60 min,the first antibody with reaction time of 90 min,HRP labeled rabbit anti-chicken IgG as second antibody with dilution of 1:3 500,reaction time of 60 min,and colour time of 10 min.The detection of 145 serum samples by iELISA and heamagglutinin inhibition(HI) showed that the coincidence of both assays was 92.41%,and ELISA sensitivity slightly higher than that of HI,with the positiverate of 92.41% and 90.34%,respectively. |
| Keywords: | avian infectious bronchitis indirect ELISA antibody detection |
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