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

• Published in: Archives of virology Vol. 161; no. 7; pp. 1957 - 1961
• Main Authors: Banerjee, Amrita, Roy, Somnath, Sharma, Susheel Kumar, Dutta, Sudip Kumar, Chandra, Satish, Ngachan, SV
• Format: Journal Article
• Language: English
• Published: Vienna Springer Vienna 01.07.2016; Springer Nature B.V
• 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; England; United Kingdom > UK; Thailand; Asia; India; Tobacco Etch Virus; Lamp Assay; Reverse Transcription Polymerase Chain Reaction; Melon Yellow Spot Virus; Chilli Plant
• ISSN: 0304-8608; 1432-8798
• DOI: 10.1007/s00705-016-2850-7

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.