禽流感病毒H5 亚型金标快速诊断技术方法的建立及试剂盒研制.pdfVIP

农业生物技术学报，2010 年，第18 卷，第2 期，第400-405 页 Journal of Agricultural Biotechnology, 2010, Vol.18, No.2, 400-405 技术创新 Updated Technology 禽流感病毒H5 亚型金标快速诊断技术方法的建立及试剂盒研制 1,2* 2,3 2,3 4 4 4 5 李克生 杜惠芬 连晓雯 徐海聂 薄清如 黄新民 才学鹏 1 甘肃农业大学, 兰州, 730070; 2 甘肃省医学科学研究院, 兰州, 730030; 3 兰州雅华生物技术有限公司, 甘肃省生物检测工程技术研究中心, 兰 州, 730030; 4 珠海出入境检验检疫局, 珠海, 519015; 5 中国农业科学院兰州兽医研究所, 兰州, 730046 * 通讯作者, lksem@ 摘 要 应用纯化禽流感病毒(AIV) H5 抗原免疫Balb/c 小鼠，得到2 株配对良好的抗AIV H5 单抗；采用金 标免疫层析法制备AIV H5 亚型金标快速诊断试剂盒，并对试剂盒性能进行评价。实验结果表明，AIV-H5 金 标试剂盒与 7 AIV H5 以外其它亚型及其它禽类病毒不发生交叉反应，其灵敏度达 1/2 ，稳定性良好；试剂盒检 测人工感染AIV H5 的鸡脏器组织，结果除心脏、泄殖腔拭子和喉拭子外，其余脏器均呈阳性，与鸡胚病毒分 离培养结果相符；田间试验结果与荧光RT-PCR 试剂盒检测符合率达 100%。 关键词 禽流感病毒, 快速诊断技术, 试剂盒 Development of Rapid Detecting Method and Lateral Flow Assay of H5 Subtype Avian Influenza Virus Li Kesheng 1,2* Du Huifen 2,3 Lian Xiaowen 2,3 Xu Hainie 4 Bo Qingru 4 Huang Xinmin 4 Cai Xuepeng 5 1 Gansu Agriculture University, Lanzhou, 730070; 2 Gansu Institute of Medical Secicne, Lanzhou, 730030; 3 Lanzhou Yahua Biotech Co.Ltd, Gansu Province Biotech and Engineeering Research Center, Lanzhou, 730030; 4 Zhuhai Entry-Exit Inspection and Quarantine Bureau, Zhuhai, 519015; 5 Lanzhou Veterinary Institute, Chinese Sciences Academy of Agriculture, Lanzhou, 730046 * Corresponding author, lksem@ DOI: 10.3969/j.issn.1674-7968.2010.02.032 Abstract Balb/c mice were immunized by purified Avian influenza virus (AIV) H5 antigen, 2 cells of monoclon- al antibody (McAb) which anti-AIV-H5 had good antigen matching, were produced. One kind of AIV-H5 lateral flow assay was developed by using gold immunochromatography, and its capability were evaluated. Experiment results showed that the AIV-H5 lateral flow assay had no cross-reaction with other substype of AIV and other fowlery viruses , and had high sensibility (1/27) and good stability.