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

Xiaohua Zou; Chao Dong; Yiduo Ni; Shidong Yuan; Qing-Hua Gao

doi:10.1016/j.jviromet.2022.114566

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

J Virol Methods. 2022 Sep:307:114566. doi: 10.1016/j.jviromet.2022.114566. Epub 2022 Jun 11.

Authors

Xiaohua Zou¹, Chao Dong², Yiduo Ni³, Shidong Yuan⁴, Qing-Hua Gao⁵

Affiliations

¹ Shanghai Key Laboratory of Protected Horticultural Technology, Forestry and Fruit Tree Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China. Electronic address: zouxh_113@126.com.
² Shanghai Key Laboratory of Protected Horticultural Technology, Forestry and Fruit Tree Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China. Electronic address: 2016204013@njau.edu.cn.
³ Shanghai Key Laboratory of Protected Horticultural Technology, Forestry and Fruit Tree Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China. Electronic address: 1131706559@qq.com.
⁴ Shanghai Key Laboratory of Protected Horticultural Technology, Forestry and Fruit Tree Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China. Electronic address: 1476723553@qq.com.
⁵ Shanghai Key Laboratory of Protected Horticultural Technology, Forestry and Fruit Tree Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China. Electronic address: qhgao20338@sina.com.

PMID: 35700833
DOI: 10.1016/j.jviromet.2022.114566

Abstract

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 ℃ 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.

Keywords: Diagnosis; RT-RPA; SMoV; Strawberry.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

China
Fragaria*
Nucleic Acid Amplification Techniques / methods
Recombinases / genetics
Reverse Transcription
Secoviridae* / genetics
Sensitivity and Specificity

Substances

Recombinases