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Reverse transcription-recombinase polymerase amplification combined with lateral flow strip for detection of rice black-streaked dwarf virus in plants

Zhao, Chong, Sun, Feng, Li, Xuejuan, Lan, Ying, Du, Linlin, Zhou, Tong, Zhou, Yijun
Journal of virological methods 2019 v.263 pp. 96-100
Laodelphax striatellus, Rice black streaked dwarf virus, complementary DNA, financial economics, plant diseases and disorders, plant viruses, reverse transcriptase polymerase chain reaction, rice, screening, temperature, China
Rice black-streaked dwarf virus (RBSDV) infects rice plants, a major crop, and is transmitted via the small brown planthopper (SBPH: Laodelphax striatellus Fallén), causing significant economic loss in China. To rapidly diagnose RBSDV, a reverse transcription-recombinase polymerase amplification (RT-RPA) method was developed using P10 virus-specific primers and probes. Detection of terminally labeled amplification products was achieved with the lateral flow strip method. Our results demonstrate that RT-RPA and RT-PCR assays offer similar sensitivity and specificity in RBSDV detection using cDNA as template. The optimum RT-RPA reaction temperature and time was 37 °C and 20 min, respectively. By screening twenty-one field suspected rice plants, the RT-RPA assay was confirmed to be simple, rapid and reliable. Thus, the RBSDV RT-RPA assay developed here will be a successful tool for quick diagnosis of RBSDV-infected rice plants.