Rapid and sensitive detection of Lily symptomless virus by reverse transcription loop-mediated isothermal amplification

• Published in: Journal of virological methods 2016-12, Vol.238, p.38-41
• Main Authors: He, Xiangfeng, Xue, Fei, Xu, Shufa, Wang, Wenhe
• Format: Article
• Language: English
• Subjects: Betaflexiviridae; Carlavirus - genetics; Carlavirus - isolation & purification; Detection; DNA Primers; Lilium - virology; Lilium spp; Limit of Detection; LSV; Nucleic Acid Amplification Techniques - economics; Nucleic Acid Amplification Techniques - methods; Plant Diseases - virology; Reverse Transcription; RNA, Viral - analysis; RT-LAMP; Sensitivity and Specificity; Temperature
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2016.10.003

•We developed a sensitive RT-LAMP assay for rapid detection of LSV.•The RT-LAMP procedure could be completed within 30min.•The detection limit for RT-LAMP was 10-fold greater than that for conventional RT-PCR. Lily symptomless virus (LSV) is one of the most prevalent viruses that infect lily plants worldwide. A rapid and sensitive reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for detection of LSV, using two primer pairs that specifically amplified the conserved sequence of LSV coat protein. The optimum reaction conditions were as follows: 4mM MgCl2 and 0.8M betaine with incubation at 64°C for 30min. The limit of detection of LSV from infected lily leaves was 10-fold higher for RT-LAMP than for conventional RT-PCR. Moreover, RT-LAMP detected LSV in not only symptomatic, but also in symptomless tissues of infected plants. These findings indicate that our RT-LAMP method for LSV can serve as a low-cost, simple, and rapid alternative to conventional detection assays.