Establishment of LAMP detection method for Rhagoletis cerasi(L.)
Establishing rapid on-site quarantine and identification methods at entry points such as air-ports,seaports,and land transport ports can effectively prevent the spread and dispersion of invasive alien pest through global commodity trade,passenger transportation,and other means,there by avoiding serious economic losses.This study designed specific LAMP primers based on the mitochondrial cy-tochrome COI gene sequence of Rhagoletis cerasi,and developed visual LAMP detection method and re-al-time fluorescent LAMP detection method.The results showed that the optimized visual LAMP detec-tion system can detect as low as 10pg/μL of R.cerasi genomic DNA at 62℃ for 40 minutes,without cross reactivity with other similar species samples.The detection results can be directly judged with the naked eye and have good specificity;The minimum detection limit of real-time fluorescent LAMP detec-tion method is 10ng/μL,and the detection time is about 90 minutes.The rapid,accurate,and sensitive visual isothermal amplification system established in this study can be applied to the on-site rapid detec-tion of R.cerasi.
万方数据
维普
