Establishment of one-step reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay for the detection of potato aucuba mosaic virus

Abstract

Potatoes, as the fourth largest global food crop, are extensively cultivated worldwide. However, potato production is significantly impacted by plant viruses, among which the potato aucuba mosaic virus (PAMV) is a notable threat. PAMV frequently co-infects with other plant viruses, severely reducing potato yields and posing a risk to the sustainable development of the potato industry. To better control the disease caused by PAMV, a practical detection method is urgently in need. In this study, we developed a one-step reverse-transcription loop-mediated isothermal amplification (one-step RT-LAMP) assay for the detection of PAMV. Primers for the one-step RT-LAMP were designed by targeting conserved regions of all PAMV isolates available in online databases, and the reaction conditions, including temperature, were optimized. Through comparative analysis with conventional RT-PCR and serological tests, we confirmed that the one-step RT-LAMP method developed here is both specific and efficient. Furthermore, the detection limit of the assay was determined to be approximately 2 copies of PAMV, demonstrating its high sensitivity. The practical applicability of the method was validated by testing potato and pepino samples collected from Gansu and Yunnan provinces, with results indicating its effectiveness in field sample detection. Overall, our findings demonstrate that the one-step RT-LAMP method is accurate, reliable, and highly practical, offering a valuable tool for controlling PAMV derived diseases and contributing to the sustainable development of the potato industry.