
CLC number: S41-30
On-line Access: 2024-08-27
Received: 2023-10-17
Revision Accepted: 2024-05-08
Crosschecked: 2019-03-02
Cited: 0
Clicked: 6461
De-Qing Huang, Rui Chen, Ya-Qin Wang, Jian Hong, Xue-Ping Zhou, Jian-Xiang Wu. Development of a colloidal gold-based immunochromatographic strip for rapid detection of Rice stripe virus[J]. Journal of Zhejiang University Science B,in press.Frontiers of Information Technology & Electronic Engineering,in press.https://doi.org/10.1631/jzus.B1800563 @article{title="Development of a colloidal gold-based immunochromatographic strip for rapid detection of Rice stripe virus", %0 Journal Article TY - JOUR
一种快速检测水稻条纹病毒的胶体金免疫层析试纸条的研制创新点:利用制备的RSV单克隆抗体,首次制备了能在5~10 min内快速、准确、灵敏、特异地检测水稻植物和灰飞虱体内RSV的胶体金免疫层析试纸条. 方法:以差速离心方法提纯的RSV病毒粒子作为免疫原免疫BALB/c小鼠,通过杂交瘤技术获得了高度特异和灵敏的RSV单克隆抗体.采用柠檬酸钠还原氯金酸的方法制备胶体金并标记一个RSV单克隆抗体,另一个RSV单克隆抗体和羊抗鼠抗体分别包被到硝酸纤维素膜的检测线和质控线,将吸水滤纸制成的样品垫、RSV免疫胶体金垫、结合有RSV单抗和羊抗鼠抗体的硝酸纤维素膜和吸水纸依次粘贴到聚氯乙烯(PVC)胶板上研制检测水稻植物和传毒介体灰飞虱中RSV的胶体金免疫层析试纸条.对田间样品进行RSV检测,分析试纸条检测RSV的有效性. 结论:利用杂交瘤技术获得了2株RSV单克隆抗体(16E6和11C1),胶体金标记16E6单克隆抗体包被在聚酯膜制成的结合垫上,以11C1单克隆抗体和羊抗鼠抗体分别包被到硝酸纤维素膜的检测线和质控线,制成能在5~10 min内快速、特异、灵敏、准确地检测田间水稻及单头灰飞虱样品中RSV的胶体金免疫层析试纸条.试纸条检测感染RSV的水稻植株和灰飞虱呈阳性反应,而检测健康水稻和感染其它5种常见水稻病毒的植株及其传毒介体呈阴性反应,且试纸条检测RSV感染水稻植物组织的灵敏度达到1:20 480倍稀释(w/v, g/mL),检测携带RSV单头灰飞虱的灵敏度达到1:2560倍稀释(单头灰飞虱/µL).田间样品检测结果发现,试纸条检测结果与反转录聚合酶链反应(RT-PCR)的检测结果一致,表明制备的试纸条可有效地用于田间RSV的检测,从而为水稻条纹叶枯病的诊断、监测预警、抗病育种、流行病学研究及科学防治提供技术和物质支撑. 关键词组: Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article
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