Specific detection and quantification of Plum pox virus by real-time fluorescent reverse transcription-PCR

Journal of Virological Methods
William L SchneiderReid D Frederick

Abstract

Plum pox virus (PPV), a destructive and economically devastating pathogen of Prunus species, was recently discovered in Pennsylvania and Canada. Current containment efforts involve eradication of infected trees based on ELISA surveys, which are laborious and less sensitive than PCR-based techniques. A real-time, fluorescent, reverse transcription-polymerase chain reaction (RT-PCR) assay was developed for the detection of PPV in the Smart Cycler (Cepheid). The methods developed are reproducible, specific to PPV, and sensitive enough to consistently detect PPV transcripts at the 10-20 fg level. The assay is more sensitive than either ELISA or traditional PCR followed by visualization with ethidium-bromide. PPV was detected from multiple hosts and from multiple Prunus tissues (leaf, stem, bud, and root). A dilution series using an in vitro synthesized transcript containing the target sequence as a standard demonstrated that the assay was effective for quantitation of viral template. The real-time PCR assay is a valuable tool for PPV detection and liter quantification in field or laboratory settings.

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Citations

Sep 2, 2005·Molecular Plant-microbe Interactions : MPMI·Jean-Michel HilyMichel Ravelonandro
May 19, 2010·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Siobain Duffy, Yee Mey Seah
Aug 17, 2006·Journal of Virology·Akhtar AliMarilyn J Roossinck
Dec 4, 2012·Viruses·Jiri SochorRene Kizek
Mar 11, 2008·Journal of Virological Methods·Fatima OsmanAdib Rowhani
Jan 16, 2007·Journal of Virological Methods·Fatima OsmanAdib Rowhani
Oct 25, 2011·Molecular Plant Pathology·Karen-Beth G ScholthofGary D Foster
Oct 10, 2013·Molecular Plant Pathology·Juan Antonio GarcíaThierry Candresse
Oct 3, 2006·Journal of Virological Methods·Aniko Varga, Delano James
Jan 1, 2006·European Journal of Plant Pathology·Rick MumfordIan Barker
Sep 18, 2007·The Journal of General Virology·Christopher M WallisWilliam L Schneider
Mar 9, 2021·Archives of Virology·Marcos D SalinaFrancisco J Reynaldi

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