Abstract
A closed-tube reverse transcription loop-mediated isothermal amplification (CT-RT-LAMP) assay was developed for the detection of yam mosaic virus (YMV, genus Potyvirus) infecting yam (Dioscorea spp.). The assay uses a set of six oligonucleotide primers targeting the YMV coat protein region, and the amplification products in YMV-positive samples are visualized by chromogenic detection with SYBR Green I dye. The CT-RT-LAMP assay detected YMV in leaf and tuber tissues of infected plants. The assay is 100 times more sensitive in detecting YMV than standard RT-PCR, while maintaining the same specificity.
MeSH terms
-
Benzothiazoles
-
Capsid Proteins / analysis*
-
Capsid Proteins / biosynthesis
-
Capsid Proteins / genetics
-
DNA Primers / chemical synthesis
-
DNA Primers / metabolism
-
Diamines
-
Dioscorea / virology*
-
Fluorescent Dyes / chemistry
-
Gene Expression
-
Nucleic Acid Amplification Techniques*
-
Organic Chemicals / chemistry
-
Plant Diseases / virology
-
Plant Leaves / virology
-
Plant Tubers / virology
-
Potyvirus / genetics*
-
Potyvirus / metabolism
-
Quinolines
-
Reverse Transcription*
-
Sensitivity and Specificity
Substances
-
Benzothiazoles
-
Capsid Proteins
-
DNA Primers
-
Diamines
-
Fluorescent Dyes
-
Organic Chemicals
-
Quinolines
-
SYBR Green I