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| H5亚型禽流感病毒反应原性差异及ELISA检测方法的研究 | |
| 引用本文： | 宋建领,张文东,张富强,李作生,冯子良,吕粤,岳亮,王金萍,胡媛媛,张应国,范泉水,邱薇.H5亚型禽流感病毒反应原性差异及ELISA检测方法的研究[J].中国兽医科学,2007,37(10):839-844. |
| 作者姓名： | 宋建领 张文东 张富强 李作生 冯子良 吕粤 岳亮 王金萍 胡媛媛 张应国 范泉水 邱薇 |
| 作者单位： | 1. 云南省热带亚热带动物病毒病重点实验室,云南,昆明,650224 2. 云南省动物疫病预防控制中心,云南,昆明,650051 3. 成都军区疾病预防控制中心,云南,昆明,650032 |
| 基金项目： | 云南省科技攻关计划;昆明市科技重点项目;云南省跨世纪人才培养基金 |
| 摘 要： | 在获得禽流感病毒多克隆抗体及H5亚型特异性单克隆抗体的基础上,建立了H5亚型特异性抗原捕捉ELISA检测方法。通过分析不同单克隆抗体与不同禽流感毒株的反应性差异,筛选高特异性和高敏感性的H5亚型单克隆抗体。优化反应条件,确定包被抗体、检测抗体及酶标抗体的最佳工作浓度,进行敏感性、特异性、重复性及稳定性分析,并与斑点ELISA、H5N1多重RT-PCR或病毒分离鉴定的结果相比较,同时使用该方法对野外样品进行了检测。结果表明,该方法敏感、特异,具有良好的重复性和稳定性,可用于检测临床样品、鸡胚培养物及细胞培养物中的H5亚型禽流感病毒。 |
| 关 键 词： | 禽流感病毒 H5亚型特异性 抗原捕捉ELISA |
| 文章编号： | 1673-4696(2007)10-0839-06 |
| 修稿时间： | 2007年6月26日 |
Analysis of viral reactogenicity difference and development of H5 subtype-specific ELISA for the detection of avian influenza virus |
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| SONG Jian-ling,ZHANG Wen-dong,ZHANG Fu-qiang,LI Zuo-sheng,FENG Zi-liang,L Yue,YUE Liang,WANG Jin-ping,HU Yuan-yuan,ZHANG Ying-guo,FAN Quan-shui,QIU Wei.Analysis of viral reactogenicity difference and development of H5 subtype-specific ELISA for the detection of avian influenza virus[J].Veterinary Science in China,2007,37(10):839-844. | |
| Authors: | SONG Jian-ling ZHANG Wen-dong ZHANG Fu-qiang LI Zuo-sheng FENG Zi-liang L Yue YUE Liang WANG Jin-ping HU Yuan-yuan ZHANG Ying-guo FAN Quan-shui QIU Wei |
| Institution: | SONG Jian-ling,ZHANG Wen-dong,ZHANG Fu-qiang,LI Zuo-sheng,FENG Zi-liang,L(U) Yue,YUE Liang,WANG Jin-ping,HU Yuan-yuan,ZHANG Ying-guo,FAN Quan-shui,QIU Wei |
| Abstract: | Based on the polyclonal antibody and H5 subtype-specific monoclonal antibody against avian influenza virus(AIV),an antigen capture ELISA(AC-ELISA) specific to H5 subtype AIV was developed.The H5 subtype-specific monoclonal antibody with high specificity and sensitivity was selected by analysis of reactivity difference between different monoclonal antibodies or different isolates of AIV.The reaction condition and the working concentrations of coating antibody,detection antibody and enzyme-labeled antibody were optimized,and the assay's sensitivity,specificity,repeatability,stability were analyzed and compared with those of Dot-ELISA,H5N1 multiplex RT-PCR and the virus isolation and identification.The results indicated that the developed AC-ELISA was sensitive,specific,and of good repeatability and stability for detection of H5 subtype AIV in clinical samples,embryo and cell culture tissues. |
| Keywords: | avian influenza virus H5 subtype-specific antigen capture ELISA |
| 本文献已被 CNKI 万方数据 等数据库收录！ | |