苜蓿花叶病毒(AMV)LAMP检测方法的建立与检验OA

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.