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基于RT-RPA和试纸条的烟草环斑病毒可视化快速检测技术研究

Visual rapid detection of tobacco ringspot virus based on reverse transcription-recombinase polymerase amplification and test strip

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【作者】 韩春雪徐司琦王欣宇谢丽杨王琰周涛范在丰

【Author】 Han Chunxue;Xu Siqi;Wang Xinyu;Xie Liyang;Wang Yan;Zhou Tao;Fan Zaifeng;Key Laboratory of Surveillance and Management for Plant Quarantine Pests-MARA,College of Plant Protection,China Agricultural University;Sanya Institute of China Agricultural University;

【通讯作者】 范在丰;

【机构】 中国农业大学植物保护学院农业农村部植物检疫性有害生物监测防控重点实验室中国农业大学三亚研究院

【摘要】 烟草环斑病毒(tobacco ringspot virus,TRSV)分布于50多个国家,是我国进境植物检疫性病毒。该病毒的寄主范围广,可侵染54科300多种植物;通过机械方式、嫁接、线虫媒介以及种子传播,对作物产量和品质时常造成严重损失。本研究选择了TRSV的外壳蛋白基因序列设计引物,建立了基于反转录(reverse transcription,RT)-重组酶聚合酶扩增(recombinase polymerase amplification,RPA)和侧流层析试纸条(lateral flow dipstick,LFD)的RT-RPA-LFD检测方法,灵敏度为486 pg/μL。验证结果表明本研究所设计的RPA引物特异性强,操作简便,适用于田间或口岸现场,利于TRSV的监测与暴发流行的早期预警。

【Abstract】 Tobacco ringspot virus(TRSV) is prevalent in over 50 countries worldwide,and has been categorized as a plant quarantine pest in China. TRSV has a wide host range,infecting over 300 plant species in 54 families. TRSV spreads through mechanical means,grafting,nematodes and seeds,posing a significant risk of epidemics that could severely impact crop yield and quality. A detection system combining reverse transcription(RT), recombinase polymerase amplification(RPA) and lateral flow dipstick(LFD),has been established using primers based on the coat protein gene sequence. This method exhibited a sensitivity of 486 pg/μL,and specificity validation demonstrated that the RPA primers designed in this study are highly specific. The method is simple to perform,thus suitable for on-site or field detection of TRSV.This approach could be of significant importance for the monitoring and early prediction of TRSV occurrence and outbreaks.

【基金】 国家重点研发计划项目(2022YFC2601500;2022YFC2601503);三亚崖州湾科技城管理局资助项目(SYND-2021-03)
  • 【分类号】S41-30
  • 【下载频次】33
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