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.