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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:
Keizerweerd, Amber T., Chandra, Amaresh, Grisham, Michael P.
Source:
Journal of Virological Methods 2015 v.212 pp. 23
ISSN:
0166-0934
Subject:
DNA primers, Sorghum (Poaceae), Sorghum mosaic virus, Sugarcane mosaic virus, coat proteins, genes, magnesium sulfate, quantitative polymerase chain reaction, reverse transcriptase polymerase chain reaction, reverse transcription, reverse transcription loop-mediated isothermal amplification, sugarcane, viruses
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 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.
Handle:
10113/63020