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

William L. Schneider, Diana J. Sherman, Andrew L. Stone, Vernon D. Damsteegt, Reid David Frederick

Research output: Contribution to journalArticle

57 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)97-105
Number of pages9
JournalJournal of Virological Methods
Volume120
Issue number1
DOIs
StatePublished - Sep 1 2004

Fingerprint

Plum Pox Virus
Reverse Transcription
Polymerase Chain Reaction
Prunus
Enzyme-Linked Immunosorbent Assay
Ethidium
Canada
Real-Time Polymerase Chain Reaction

All Science Journal Classification (ASJC) codes

  • Virology

Cite this

Schneider, William L. ; Sherman, Diana J. ; Stone, Andrew L. ; Damsteegt, Vernon D. ; Frederick, Reid David. / Specific detection and quantification of Plum pox virus by real-time fluorescent reverse transcription-PCR. In: Journal of Virological Methods. 2004 ; Vol. 120, No. 1. pp. 97-105.
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Specific detection and quantification of Plum pox virus by real-time fluorescent reverse transcription-PCR. / Schneider, William L.; Sherman, Diana J.; Stone, Andrew L.; Damsteegt, Vernon D.; Frederick, Reid David.

In: Journal of Virological Methods, Vol. 120, No. 1, 01.09.2004, p. 97-105.

Research output: Contribution to journalArticle

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