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| 免疫PCR检测微量H5亚型禽流感病毒 | |
| 引用本文： | 邓明俊,肖西志,吴振兴,张彦明,辛学谦,郑小龙,王群,王昌军.免疫PCR检测微量H5亚型禽流感病毒[J].畜牧兽医学报,2011,42(7). |
| 作者姓名： | 邓明俊 肖西志 吴振兴 张彦明 辛学谦 郑小龙 王群 王昌军 |
| 作者单位： | 1. 西北农林科技大学动物医学院,杨凌712100;山东出入境检验检疫局,青岛266002 2. 山东出入境检验检疫局,青岛,266002 3. 西北农林科技大学动物医学院,杨凌,712100 |
| 摘 要： | 为了有效检测低浓度禽流感病毒,笔者通过将高特异性的抗原-抗体反应与高灵敏性的PCR扩增技术相结合,建立了一种快速检测痕量H5亚型禽流感病毒的间接免疫PCR方法和间接“三明治”抗体夹心免疫PCR方法.以TopYield 96孔板为固相免疫吸附载体,以禽流感病毒H5蛋白为检测对象,通过一系列免疫反应及亲和素-生物素桥联作用,将生物素标记的报告DNA分子和生物素标记的禽流感病毒H5亚型抗体分子连接.充分洗涤后,在TopYield板孔中添加PCR反应液,对板壁偶联的报告DNA进行PCR扩增,间接达到检测微量禽流感病毒H5蛋白的目标.通过优化报告DNA分子和链亲和素的浓度,2种免疫PCR技术都可检测到约1 fg的禽流感病毒H5蛋白,与常规ELISA方法相比,灵敏性提高了近1 000倍. |
| 关 键 词： | 免疫PCR 禽流感病毒 H5血凝素蛋白 |
Immuno-PCR Assay for Detection of H5 Hemagglutinin Subtype Avian Influenza Virus |
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| DENG Ming-jun , XIAO Xi-zhi , WU Zhen-xing , ZHANG Yan-ming , XIN Xue-qian , ZHENG Xiao-long , WANG Qun , WANG Chang-jun.Immuno-PCR Assay for Detection of H5 Hemagglutinin Subtype Avian Influenza Virus[J].Acta Veterinaria et Zootechnica Sinica,2011,42(7). | |
| Authors: | DENG Ming-jun XIAO Xi-zhi WU Zhen-xing ZHANG Yan-ming XIN Xue-qian ZHENG Xiao-long WANG Qun WANG Chang-jun |
| Institution: | DENG Ming-jun1,2,XIAO Xi-zhi1,WU Zhen-xing2,ZHANG Yan-ming1,XIN Xue-qian2,ZHENG Xiao-long2,WANG Qun2,WANG Chang-jun2(1.Northwest Agriculture and Forestry University,College of Veterinary Medicine,Yangling 712100,China,2.Shandong Entry-Exit Inspection and Quarantine Bureau,Qingdao 266002,China) |
| Abstract: | In order to detect the low concentration of avian influenza virus(AIV) in the tracheal and cloacal swabs taken from the HPAI-infected poultry,an indirect immuno-polymerase chain reaction(immuno-PCR) assay and an indirect sandwich immuno-PCR assay,based on Immunology and polymerase chain reaction(IPCR) which combining enzyme immunoassay with DNA amplification,were developed.For the indirect immuno-PCR assay,TopYield microtiter plates were coated with purified AIV/H5 protein that was recognized by anti-AIV/H5... |
| Keywords: | immuno-PCR avian influenza virus H5 hemagglutinin protein |
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