Short communicationRapid detection of Prunus necrotic ringspot virus using magnetic nanoparticle-assisted reverse transcription loop-mediated isothermal amplification

•A new method to detect Prunus necrotic ringspot virus (PNRSV) from cherry trees was developed using reverse transcription loop-mediated isothermal amplification (RT-LAMP).•The data demonstrated that RT-LAMP is more sensitive than RT-PCR for detection of PNRSV.•This method is less expensive in terms of the equipment required and highly specific for PNRSV detection.•Improvement on total RNA extraction using the magnetic nanoparticles method may shorten the experiment time and achieve ideal results.•To our knowledge, this is the first report on developing a RT-LAMP assay to detect PNRSV in fruit trees.

Prunus necrotic ringspot virus (PNRSV) has seriously reduced the yield of Prunus species worldwide. In this study, a highly efficient and specific two-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed to detect PNRSV. Total RNA was extracted from sweet cherry leaf samples using a commercial kit based on a magnetic nanoparticle technique. Transcripts were used as the templates for the assay. The results of this assay can be detected using agarose gel electrophoresis or by assessing in-tube fluorescence after adding SYBR Green I. The assay is highly specific for PNRSV, and it is more sensitive than reverse-transcription polymerase chain reaction (RT-PCR). Restriction enzyme digestion verified further the reliability of this RT-LAMP assay. To our knowledge, this is the first report of the application of RT-LAMP to PNRSV detection in Prunus species.