Record Details

Rapid detection of Piper yellow mottle virus and Cucumber mosaic virusinfecting black pepper (Piper nigrum) by loop-mediated isothermalamplification (LAMP)

DSpice at Indian Institute of Spices Research

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Field Value
 
Creator BHAT, A I
SILJO, A
DEESHMA, K P
 
Date 2014-06-05T05:35:49Z
2014-06-05T05:35:49Z
2013
 
Identifier Journal of Virological Methods 193:190-196 (2013)
http://hdl.handle.net/123456789/1116
 
Description The loop-mediated isothermal amplification (LAMP) assay for Piper yellow mottle virus and the reverse transcription (RT) LAMP assay for Cucumber mosaic virus each consisted of a set of five primers designed against the conserved sequences in the viral genome. Both RNA and DNA isolated from black pepper were used as a template for the assay. The results were assessed visually by checking turbidity, green fluorescence and pellet formation in the reaction tube and also by gel electrophoresis. The assay success-fully detected both viruses in infected plants whereas no cross-reactions were recorded with healthy plants. Optimum conditions for successful amplification were determined in terms of the concentrations of magnesium sulphate and betaine, temperature, and duration. The detection limit for both LAMP and RT-LAMP was up to 100 times that for conventional PCR and up to one-hundredth of that for real-time PCR. The optimal conditions arrived at were validated by testing field samples of infected vines of three species from different regions.
 
Format 103273 bytes
application/pdf
 
Language en
 
Publisher Elsevier
 
Subject LAMP
PCR
real-time PCR
Sensitivity
Validation
 
Title Rapid detection of Piper yellow mottle virus and Cucumber mosaic virusinfecting black pepper (Piper nigrum) by loop-mediated isothermalamplification (LAMP)
 
Type Article