Location: Sugarcane ResearchTitle: Development of a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the detection of Sugarcane mosaic virus and Sorghum mosaic virus in sugarcane Author
|Chandra, Amaresh - Indian Institute Of Sugarcane Research|
Submitted to: Journal of Virological Methods
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/21/2014
Publication Date: 11/21/2014
Publication URL: http://handle.nal.usda.gov/10113/63020
Citation: Keizerweerd, A.T., Chandra, A., Grisham, M.P. 2014. Development of a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the detection of Sugarcane mosaic virus and Sorghum mosaic virus in sugarcane. Journal of Virological Methods. 212(2015):23-29. DOI: 10.1016/j.jviromet.2014.10.013
Interpretive Summary: This work presents a diagnostic method known as RT-LAMP that is capable of detecting two viral pathogens in sugarcane. Its simplicity, rapidity, and low cost make it more advantageous than other frequently applied techniques. This research demonstrates our success in not only detecting the closely related Sugarcane mosaic virus (SCMV) and Sorghum mosaic virus (SrMV), but also in discriminating between them in field samples. Moreover, our method was able to detect all strains of each virus commonly found in Louisiana sugarcane. This approach can potentially be applied to crop analysis where rapid pathogen detection is desirable in field laboratories with limited resources.
Technical Abstract: A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for detecting Sugarcane mosaic virus (SCMV) and Sorghum mosaic virus (SrMV) in sugarcane. Six sets of four primers corresponding to the conserved coat protein gene were designed for each virus and their success at amplifying their targets was tested. Results were obtained by visually inspecting reaction color and gel electrophoresis products. All primer sets that worked were also shown to be specific for their target virus. In addition, several strains of each virus were detectable with the chosen primers. Magnesium sulphate concentrations were optimized, with both viruses requiring a minimum of 5 mM for detection. The sensitivity of our RT-LAMP assay, however, was less than that of conventional and real-time RT-PCR. This is the first report of an RT-LAMP assay for SCMV and SrMV detection in sugarcane.