Development of a colloidal gold-based immunochromatographic strip for rapid detection of Rice stripe virus
Development of a colloidal gold-based immunochromatographic strip for rapid detection of Rice stripe virus
复制标题
水稻条纹病毒快速检测胶体金免疫层析试纸条的研制
DOI：
10.1631/jzus.b1800563
复制
发表时间：
2019-04-01
影响因子：
5.1
通讯作者：
Wu, Jian-xiang
中科院分区：
文献类型：
--
作者：
Huang, De-qing;Chen, Rui;Wu, Jian-xiang
水稻条纹病毒（RSV）在全球水稻生产中造成巨大损失。本研究建立了抗RSV的单克隆抗体16E6和11C1，以及基于胶体金的免疫层析试纸条，用于水稻和飞虱中RSV的特异、灵敏、快速检测。用胶体金偶联MAb 16E6，将MAb 11C1包被在试纸的硝化纤维素膜的测试线上。对rsv侵染水稻植株和小褐飞虱（SBPH）呈阳性反应，对其他5种水稻病毒、健康水稻植株、5种水稻病毒载体和非毒性SBPH呈阴性反应，证实了试纸条的特异性。灵敏度分析表明，该试纸条在rsv感染的水稻组织粗提物（w/v, g/mL）和毒力强的SBPH匀浆（单个SPBH/µL）中均能检测到该病毒。利用田间采集的水稻和SBPH样品进一步验证了该条带的有效性。该试纸条是一种低成本、快速、易用的现场检测工具，可用于田间流行病学研究和水田调查中RSV感染的现场检测，为田间RSV管理提供决策依据。
Rice stripe virus (RSV) causes dramatic losses in rice production worldwide. In this study, two monoclonal antibodies (MAbs) 16E6 and 11C1 against RSV and a colloidal gold-based immunochromatographic strip were developed for specific, sensitive, and rapid detection of RSV in rice plant and planthopper samples. The MAb 16E6 was conjugated with colloidal gold and the MAb 11C1 was coated on the test line of the nitrocellulose membrane of the test strip. The specificity of the test strip was confirmed by a positive reaction to RSV-infected rice plants and small brown planthopper (SBPH), and negative reactions to five other rice viruses, healthy rice plants, four other vectors of five rice viruses, and non-viruliferous SBPH. Sensitivity analyses showed that the test strip could detect the virus in RSV-infected rice plant tissue crude extracts diluted to 1:20480 (w/v, g/mL), and in individual viruliferous SBPH homogenate diluted to 1:2560 (individual SPBH/µL). The validity of the developed strip was further confirmed by tests using field-collected rice and SBPH samples. This newly developed test strip is a low-cost, fast, and easy-to-use tool for on-site detection of RSV infection during field epidemiological studies and paddy field surveys, and thus can benefit decision-making for RSV management in the field.