A reverse transcription loop‐mediated isothermal amplification assay for the detection of strawberry mottle virus. Issue 5 (7th February 2021)

• Record Type: Journal Article
• Authors: Ren, Junda; Wang, Jia; Zhu, Yadong; Han, Chenggui; Shang, Qiaoxia
• Is Part Of: Journal of phytopathology. Volume 169:Issue 5(2021)
• Page Start: 295
• Page End: 302
• Publication Date: 2021-02-07
• Subjects: detection -- RT‐LAMP -- RT‐PCR -- strawberry mottle virus; Plant diseases -- Periodicals; 632
• DOI: 10.1111/jph.12985
• Languages: English
• ISSNs: 0931-1785

Abstract

Abstract: Strawberry mottle virus (SMoV) is one of the most important viruses infecting strawberry all over the world, seriously affecting the yield and quality of strawberry especially in the occasion of compound infection. Loop‐mediated isothermal amplification (LAMP) as a new method of nucleic acid amplification has the advantages of rapid detection, simple operation and high sensitivity. Therefore, four specific LAMP primers using the 3′ end conservative coat protein (CP) gene were selected and designed according to the RNA2 sequence of SMoV obtained in NCBI GenBank. Results showed that the optimized RT‐LAMP reaction conditions including 1.0 µmol/L 1 primers FIP/BIP, 0.1 µmol/L 1 primers F3 /B3, 2 mmol/L 1 Mg 2+, 1.2 mmol/L 1 dNTPs, 1.0 mol/L 1 Betaine and 2 mol/L 1 DTT, with an optimal temperature set at 62°C for 45 min. The amplicons were successfully inspected by visual assessment using SYBR Green I as well as electrophoresis on agarose gel. The sensitivity of the LAMP was 100 times higher than the RT‐PCR method. Furthermore, it was highly specific for SMoV and showed no cross‐reaction with several other strawberry viruses. This assay will provide a practical method for rapid detection of large quantities of field samples and be useful in reducing the virus incidence through production of virus‐free planting material.