| 柞蚕微孢子虫胶体金免疫层析检测法 |
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| 引用本文： | 王伯阳,姜义仁,臧敏,石生林,杨瑞生,包臣,秦利.柞蚕微孢子虫胶体金免疫层析检测法[J].蚕业科学,2012,38(1):97-101. |
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| 作者姓名： | 王伯阳 姜义仁 臧敏 石生林 杨瑞生 包臣 秦利 |
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| 作者单位： | 沈阳农业大学生物科学技术学院,辽宁省昆虫资源工程技术研究中心,沈阳110866;沈阳农业大学生物科学技术学院,辽宁省昆虫资源工程技术研究中心,沈阳110866;沈阳农业大学植物保护学院,沈阳110866;吉林省农村工作委员会园艺特产站,长春,130000 |
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| 基金项目： | 现代农业产业技术体系专项，辽宁省教育厅科研项目，沈阳农业大学校青年基金 |
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| 摘 要： | 利用柞蚕微孢子虫孢子液直接免疫家兔获得柞蚕微孢子虫多克隆抗体后,分别采用双抗夹心法和竞争法制作胶体金免疫层析试纸条,建立能简便、快捷、准确诊断柞蚕微粒子病的柞蚕微孢子虫胶体金免疫层析检测法。双抗夹心法和竞争法分别是将柞蚕微孢子虫多克隆抗体与25 nm和17 nm的胶体金颗粒结合并固定在金标垫上,然后均将羊抗兔二抗包被在硝酸纤维素膜上作为质控点(C),但是2种方法制作的胶体金免疫层析试纸条的反应模式不同:前者是以柞蚕微孢子虫多克隆抗体作为检测点(T),而后者以柞蚕微孢子虫孢壁蛋白作为检测点(T)。2种方法制作的胶体金免疫层析试纸条的检测时间均为10 min,检测灵敏度为0.8×107个/mL,与柞蚕血淋巴无交叉反应,特异性强,操作简单,其中,以双抗夹心法制作的试纸条显色更清晰,结果更可靠,更具实用价值。
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| 关 键 词： | 柞蚕微孢子虫 多克隆抗体 胶体金 免疫层析 双抗夹心法 竞争法 |
Rapid Detection of Nosema pernyi by Colloidal Gold Immunochromatography |
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| WANG Bo-Yang
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JIANG Yi-Ren
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ZANG Min
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SHI Sheng-Lin
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YANG Rui-Sheng
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BAO Chen
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QIN Li.Rapid Detection of Nosema pernyi by Colloidal Gold Immunochromatography[J].Acta Sericologica Sinica,2012,38(1):97-101. |
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| Authors: | WANG Bo-Yang JIANG Yi-Ren ZANG Min SHI Sheng-Lin YANG Rui-Sheng BAO Chen QIN Li |
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| Institution: | 1(1College of Bioscience and Biotechnology,Shenyang Agricultural University,Liaoning Engineering and Technology Research Center for Insect Resources,Shenyang 110866,China;2Horticulture and Native Product Station,Rural Work Committee of Jilin Province,Changchun 130000,China;3College of Plant Protection,Shenyang Agricultural University,Shenyang 110866,China) |
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| Abstract: | The polyclonal antibody against Nosema pernyi was prepared by immunizing rabbits using the suspension containing spores of N.pernyi and was used to prepare colloidal gold test strips for immunochromatographic assay through double antibody sandwich method and competition method.The established colloidal gold immunochromatographic assay could have a simple,rapid and accurate diagnosis to pebrine disease in Antheraea pernyi.The double antibody sandwich method and competition method conjugated the polyclonal antibody with 25 nm and 17 nm colloidal gold particles respectively and bound the conjugated products on a gold-labeled pad.Then,goat anti-rabbit secondary antibody was spotted on nitrocellulose membrane as control dot(C) for both methods.However,the reaction patterns for preparing colloidal gold test strips were different for the two methods: the former took the polyclonal antibody against N.pernyi as the test dot(T),and the latter took spore wall proteins of N.pernyi as the test dot(T).The test strips prepared from both methods had a working time of 10 min and a diagnostic sensitivity of 0.8×107 per mL,and had no cross reaction with the hemolymph of Antheraea pernyi,being highly specific and easy to operate.Besides,the test strip from double antibody sandwich method had clearer staining result and is thus more reliable and valuable for practical application. |
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| Keywords: | Nosema pernyi Polyclonal antibody Colloidal gold Immunochromatography Double antibody sandwich method Competition method |
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