南京农业大学学报2025，Vol.48Issue(6)：1309-1315,7.DOI:10.7685/jnau.202407016
基于重组聚合酶扩增技术的新德里番茄曲叶病毒快速检测方法的建立
Establishment of a fast detection method for tomato leaf curl New Delhi virus by recombinase polymerase amplification
摘要
Abstract
[Objectives]Tomato leaf curl New Delhi virus(ToLCNDV)is a significant challenge to the production of Solanaceae and Cucurbitaceae crops in China.This study aimed to develop a rapid method for the virus to facilitate disease monitoring,prevention and control.[Methods]Two pairs of specific primers were designed for the conserved region of the ToLCNDV genome A.After screening,one pair of primers were selected for the development of a recombinase polymerase amplification(RPA)assay for ToLCNDV detection.[Results]The method could achieve exponential amplification of ToLCNDV in only 30 min at a constant temperature of 42℃.The amplified products could be visualized by both agarose gel electrophoresis and flow chromatography test strips.The RPA technique developed in this study was found to be able to specifically detect ToLCNDV,but not other viruses infecting Solanaceae and Cucurbitaceae crops,such as tomato yellow leaf curl virus(TYLCV),tomato golden mosaic virus(TGMV),tomato leaf curl Yunnan virus(TLCYnV)and squash leaf curl China virus(SLCCNV).In addition,the sensitivity of the RPA technique was determined by testing its ability to detect positive results when the template DNA concentration was diluted to 5×10-7 ng.This was in contrast to the minimum template content for regular polymerase chain reaction(PCR)detection,which was only 0.05 ng.This indicated that the sensitivity of RPA for detecting ToLCNDV was significantly higher than that of regular PCR.[Conclusions]The rapid method detection for ToLCNDV developed in this study would facilitate the development of a technical system for the early monitoring and warning,comprehensive prevention and control of this virus.关键词
新德里番茄曲叶病毒/重组聚合酶扩增技术/快速检测/早期监测预警Key words
tomato leaf curl New Delhi virus/recombinase polymerase amplification/rapid detection/early monitoring and warning分类
植物保护学引用本文复制引用
植爱萍,柳勤海,赵紫馨,肖轩仪,张洋,陈莉,冯明峰,朱敏,陶小荣..基于重组聚合酶扩增技术的新德里番茄曲叶病毒快速检测方法的建立[J].南京农业大学学报,2025,48(6):1309-1315,7.基金项目
国家重点研发计划项目(2022YFD1401200) （2022YFD1401200）
南京农业大学大学生创新创业训练计划项目植物生产中心专项(ZKF202306) （ZKF202306）
