Rapid Detection of Strawberry Mild Yellow Edge Virus with a Lateral Flow Strip Reverse Transcription Recombinase Polymerase Amplification Assay.

Strawberry mild yellow edge virus (SMYEV) is a latent virus that severely affects the yield and quality of strawberry fruit. The technology suitable for rapid and accurate detection of SMYEV on site is important to effectively control its spread. In this study, a reverse transcription recombinase polymerase amplification combined with lateral flow strip (SMYEV-RT-RPA-LF), targeting the conserved genome of Beijing SMYEV isolate, was established to diagnose SMYEV in strawberries. The SMYEV-RT-RPA-LF assay showed no cross-reaction with other strawberry viruses. The sensitivity of SMYEV-RT-RPA-L assay was 100 times higher than that of RT-PCR (10 pg/µL). In addition, through the detection of suspected samples in the field, it was found that the accuracy of SMYEV-RT-RPA-L assay was consistent with the RT-PCR results. However, compared with RT-PCR, SMYEV-RT-RPA-LF assay has the advantages of simple operation, time savings, and high specificity and sensitivity, indicating the potential application of SMYEV-RT-RPA-LF in the rapid field diagnosis of SMYEV.

Rapid detection of strawberry mottle virus using reverse transcription recombinase polymerase amplification with lateral flow strip.

Strawberry mottle virus (SMoV) is one of the main RNA viruses that profoundly affects the growth of strawberries worldwide. The rapid on-site detection of SMoV described here can be applied to produce virus-free strawberry seedlings. Reverse transcriptase recombinase polymerase amplification (RT-RPA) was combined with lateral flow (LF) strip to rapidly detect SMoV. The detection limit was 500 fg of RNA under optimized conditions. The SMoV-RT-RPA-LF assay was optimal with a combination of 2 µL reverse primer (5 µM) and 0.6 µL probe (10 µM) in a 50 µL RT-RPA reaction mixture for isothermal amplification at 40 â„ƒ for 15 min. In addition, 100 suspected samples were collected from different regions in the Shanghai suburbs. The SMoV-RT-RPA-LF assay showed that 3 of these 100 samples were positive for SMoV, which was in good concordance with the reverse transcription polymerase chain reaction (RT-PCR) results. The primers and probe had a unique specificity to SMoV because there was no cross-reaction with other strawberry viruses. This study provides an effective technique for the rapid on-site detection of SMoV to ensure a virus-free strawberry nursery.