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鹅细小病毒胶体金免疫层析检测方法的建立

Establishment of colloidal gold immunochromatographic assay for detection of goose parvovirus

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【作者】 邵周伍林; 白小飞; 刘明; 张云;

【Author】 SHAOZHOU Wu-lin;BAI Xiao-fei;LIU Ming;ZHANG Yun;State Key Laboratory of Veterinary Biotechnology/Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences;

【机构】 中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室;

【摘要】 为建立一种快速、简便地检测鹅细小病毒(GPV)的方法,采用柠檬酸三钠还原法制备胶体金,将其标记于抗GPV VP3蛋白单克隆抗体2D5上,在硝酸纤维素膜上喷涂抗GPV VP3蛋白单克隆抗体4A8和山羊抗小鼠IgG抗体作为检测线和质控线,制成检测GPV的胶体金免疫层析试纸条。结果显示,该试纸条检测GPV的最低TCID50为1×104.15/mL,用制备的试纸条仅检测GPV为阳性,而检测其他主要水禽病的病原结果均为阴性;同一批次和不同批次的试纸条对GPV阳性样品的检测结果均无差异。使用该试纸条对已知的80份粪便样品进行检测,阳性及阴性样品的检测符合率分别为96.49%和100%。结果表明,制备的试纸条具有良好的特异性、敏感性和重复性,为"小鹅瘟"的快速诊断提供了新的方法。

【Abstract】 To develop a rapid and simple method for detection of goose parvovirus(GPV),a monoclonal antibody(McAb)2D5against GPV VP3 protein was labeled with colloidal gold particles using the colloidal gold immunochromatographic strip.The McAb 4A8 against GPV VP3 protein and goat anti-mouse IgG antibody were blotted on the nitrocellulose membrane as the test line and control line,respectively.The GPV detection limit was 1×104.15/mL in TCID50.And the strips did not react with other known pathogens,indicating that the strips are specific for detecting GPV.The results of identical batch and different batches of strips detecting 5positive samples were same.The corresponding rate of strips for detecting positive and negative samples was 96.49%and 100%,respectively.The established strip has a good specificity,sensitivity,and reproducibility,which provides a new technique for rapidly detecting goose parvovirus infection.

【基金】 现代农业水禽产业技术体系岗位科学家专项(CARS-43-10);公益性行业(农业)科研专项(201003012)
  • 【文献出处】 中国兽医科学 ,Chinese Veterinary Science , 编辑部邮箱 ,2015年05期
  • 【分类号】S852.65
  • 【被引频次】11
  • 【下载频次】148
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