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

•An RT-LAMP assay for Sugarcane mosaic virus and Sorghum mosaic virus was developed.•Six primer sets consisting of four primers each were designed for each virus.•All sets yielding successful amplification were specific for their target virus.•These sets were able to detect all tested strains of the...

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Veröffentlicht in:Journal of virological methods 2015-02, Vol.212, p.23-29
Hauptverfasser: Keizerweerd, Amber T., Chandra, Amaresh, Grisham, Michael P.
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Sprache:eng
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Zusammenfassung:•An RT-LAMP assay for Sugarcane mosaic virus and Sorghum mosaic virus was developed.•Six primer sets consisting of four primers each were designed for each virus.•All sets yielding successful amplification were specific for their target virus.•These sets were able to detect all tested strains of the target virus in sugarcane. 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 and tested for each virus. Three primer sets designed for detecting SCMV and four for detecting SrMV were successful in the RT-LAMP assay. The effective primer sets were not only specific for their target virus, but also able to detect multiple virus strains. The magnesium sulfate concentration of the reaction solution was optimized, with both viruses requiring a minimum of 5mM for detection. The sensitivity of this RT-LAMP assay was less than that of conventional and real-time RT-PCR.
ISSN:0166-0934
1879-0984
DOI:10.1016/j.jviromet.2014.10.013