Reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid diagnosis of chilli veinal mottle virus

Chilli veinal mottle virus (ChiVMV) causes significant economic loss to chilli cultivation in northeastern India, as well as in eastern Asia. In this study, we have developed a single-tube one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid, sensitive and...

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Bibliographic Details
Published in:Archives of virology 2016-07, Vol.161 (7), p.1957-1961
Main Authors: Banerjee, Amrita, Roy, Somnath, Sharma, Susheel Kumar, Dutta, Sudip Kumar, Chandra, Satish, Ngachan, SV
Format: Article
Language:English
Subjects:
Biomedical and Life Sciences
Biomedicine
Brief Report
Capsicum - virology
Chilli veinal mottle virus
Design
DNA Primers - genetics
Flowers & plants
Infectious Diseases
Insects
Medical Microbiology
Nucleic Acid Amplification Techniques - methods
Plant Diseases - virology
Polymerase chain reaction
Potyvirus - classification
Potyvirus - genetics
Potyvirus - immunology
Potyvirus - isolation & purification
Proteins
Reverse Transcription
Sensitivity and Specificity
Viral Proteins - genetics
Virology
Viruses
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Summary:Chilli veinal mottle virus (ChiVMV) causes significant economic loss to chilli cultivation in northeastern India, as well as in eastern Asia. In this study, we have developed a single-tube one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid, sensitive and specific diagnosis of ChiVMV. Amplification could be visualized after adding SYBR Green I (1000×) dye within 60 min under isothermal conditions at 63 °C, with a set of four primers designed based on the large nuclear inclusion protein (NIb) domain of ChiVMV (isolate KC-ML1). The RT-LAMP method was 100 times more sensitive than one-step reverse transcription polymerase chain reaction (RT-PCR), with a detection limit of 0.0001 ng of total RNA per reaction.
ISSN:0304-8608
1432-8798
DOI:10.1007/s00705-016-2850-7