CLC number: S41-30
On-line Access: 2020-10-12
Received: 2020-05-17
Revision Accepted: 2020-07-30
Crosschecked: 2020-09-08
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Citations: Bibtex RefMan EndNote GB/T7714
Wan-qin He, Jia-yu Wu, Yi-yi Ren, Xue-ping Zhou, Song-bai Zhang, Ya-juan Qian, Fang-fang Li, Jian-xiang Wu. Highly sensitive serological approaches for Pepino mosaic virus detection[J]. Journal of Zhejiang University Science B,in press.Frontiers of Information Technology & Electronic Engineering,in press.https://doi.org/10.1631/jzus.B2000255 @article{title="Highly sensitive serological approaches for Pepino mosaic virus detection", %0 Journal Article TY - JOUR
凤果花叶病毒高灵敏度血清学检测技术创新点:首次制备了抗PepMV的高度特异和灵敏的单克隆抗体,并利用制备的单抗建立了3种能特异且灵敏地检测PepMV的血清学方法. 方法:以差速离心方法提纯的PepMV粒子作为免疫原免疫BALB/c小鼠,通过杂交瘤技术获得了能稳定传代并分泌PepMV单克隆抗体的杂交瘤细胞株;杂交瘤细胞注射到小鼠腹腔获得单克隆抗体腹水,并以制备单抗为核心,根据血清学原理建立检测植物中PepMV的双抗夹心酶联免疫吸附试验(DAS-ELISA)、斑点酶联免疫吸附试验(Dot-ELISA)和组织印迹酶联免疫吸附试验(Tissue print-ELISA)三种血清学检测方法;利用田间番茄样品分析建立的血清学方法检测PepMV的有效性. 结论:利用杂交瘤技术获得了6株能分泌高度特异灵敏PepMV单克隆抗体的杂交瘤细胞株,以分泌的单抗为核心建立了检测植株中PepMV的DAS-ELISA、Dot-ELISA和Tissue print-ELISA三种高度灵敏的血清学新技术.三种建立的血清学技术检测感染PepMV的番茄植株均呈强阳性反应,而检测健康番茄及感染其他5种植物病毒的植株呈阴性反应,且DAS-ELISA和Dot-ELISA血清学技术检测番茄病叶粗提液的灵敏度分别达到1:1 310 720和1:20 480倍稀释(质量体积比,g/mL).田间样品检测结果发现,建立的血清学技术的检测结果与反转录聚合酶链反应(RT-PCR)的检测结果一致,表明建立的血清学方法可有效地用于植物中PepMV的检测.同时,本研究首次发现PepMV已在我国云南番茄作物上发生流行.PepMV单克隆抗体的制备及其灵敏血清学检测方法的建立有益于PepMV的田间调查和诊断及其科学防控. 关键词组: Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article
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