A single-step reverse transcription loop-mediated isothermal amplification assay for rapid and accurate detection of Pepper vein yellows virus

Pepper vein yellows virus (PeVYV) is a member of Polerovirus and infects solanaceous crops worldwide. In this study, a highly efficient and easy-to-use single-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for PeVYV detection. This assay used two outer (F3 and B3), two inner (FIP and BIP) and two loop (LF and LB) primers in a single reaction. The optimized reaction was determined to be at 62°C for 45 min and the whole reaction took less than 1 h. This assay was about 10 times more sensitive than the conventional one-step RT-PCR. This RT-LAMP assay should allow specific detection of PeVYV in plant tissues. We recommend this PeVYV-specific RT-LAMP assay for large scale field studies of PeVYV infection.