Chromogenic detection of yam mosaic virus by closed-tube reverse transcription loop-mediated isothermal amplification (CT-RT-LAMP)

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 th...

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Bibliographic Details
Published in:Archives of virology 2018-04, Vol.163 (4), p.1057-1061
Main Authors: Nkere, Chukwuemeka K., Oyekanmi, Joshua O., Silva, Gonçalo, Bömer, Moritz, Atiri, Gabriel I., Onyeka, Joseph, Maroya, Norbert G., Seal, Susan E., Kumar, P. Lava
Format: Article
Language:English
Subjects:
Biomedical and Life Sciences
Biomedicine
Brief Report
Capsid Proteins - analysis
Capsid Proteins - biosynthesis
Capsid Proteins - genetics
Coat protein
coat proteins
Dioscorea
Dioscorea - virology
DNA polymerase
DNA primers
DNA Primers - chemical synthesis
DNA Primers - metabolism
Flowers & plants
Fluorescent Dyes - chemistry
Gene Expression
Infectious Diseases
leaves
Medical Microbiology
Nucleic Acid Amplification Techniques
Oligonucleotides
Organic Chemicals - chemistry
Plant Diseases - virology
Plant Leaves - virology
Plant Tubers - virology
Polymerase chain reaction
Potyvirus - genetics
Potyvirus - metabolism
Primers
Proteins
reverse transcriptase polymerase chain reaction
Reverse Transcription
reverse transcription loop-mediated isothermal amplification
Sensitivity and Specificity
tubers
Virology
Viruses
Yam mosaic virus
yams
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Description
Summary: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.
ISSN:0304-8608
1432-8798
DOI:10.1007/s00705-018-3706-0