现代农业研究2026，Vol.32Issue(5)：43-48,6.
苜蓿花叶病毒(AMV)LAMP检测方法的建立与检验
Establishment and Validation of a LAMP Detection Method for Alfalfa Mosaic Virus(AMV)
摘要
Abstract
To establish a rapid,sensitive,and visual detection method for Alfalfa mosaic virus(AMV),specific LAMP primers were designed based on highly conserved sequences of the AMV coat protein(CP)gene.Through primer screening and reaction condition optimization,a visual LAMP detection system based on SYBR Green Ⅰ dye was devel-oped,and its specificity,sensitivity,and field application efficacy were validated.The results indicated that the opti-mized reaction conditions were as follows:a 60-minute isothermal reaction at 65 ℃,an internal-to-external primer con-centration ratio of 8∶1,a dNTP concentration of 0.8 mmol/L,and a Mg2+concentration of 6.0 mmol/L.The system pro-duced specific amplification for AMV and showed no cross-reaction with Alfalfa Dwarfing Virus(ADV)or Bean Leaf Roll Virus(BLRV),demonstrating excellent specificity.Sensitivity testing revealed that the method's lower limit of de-tection was 50 pg/μL,representing a 10-fold increase in sensitivity compared to conventional RT-PCR.When applied to 20 field samples suspected of infection,the positive detection rate of the established method was 85％,showing high consistency with RT-PCR validation results.This detection method is simple to operate,rapid,efficient,and cost-effec-tive,requiring no complex instrumentation.It is suitable for rapid testing in grassroots laboratories and field settings,providing reliable technical support for early warning of AMV,seedling quarantine,and disease control.关键词
紫花苜蓿/苜蓿花叶病毒(AMV)/环介导等温扩增(LAMP)/可视化检测Key words
alfalfa/alfalfa mosaic virus(AMV)/loop-mediated isothermal amplification(LAMP)/visual detection分类
农业科技引用本文复制引用
党开祥,李克梅..苜蓿花叶病毒(AMV)LAMP检测方法的建立与检验[J].现代农业研究,2026,32(5):43-48,6.基金项目
国家重点研发计划课题(项目编号:2022YFD1401101) （项目编号:2022YFD1401101）
国家自然科学基金项目(项目编号:31760708). （项目编号:31760708）
