Detection and quantification of Plum pox potyvirus in aphid vectors by real-time fluorescent reverse transcription-PCR

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

Research output: Chapter in Book/Report/Conference proceedingConference contribution

3 Scopus citations

Abstract

A real-time, fluorescent, reverse transcription-polymerase chain reaction(RT-PCR) assay was developed for the detection of Plum pox potyvirus (PPV). The methods developed are reproducible, specific to PPV, and sensitive enough to consistently detect PPV transcripts at the 10 fg level. The assay is more sensitive than either ELISA or traditional PCR followed by electrophoretic visualization with ethidium-bromide. All strains of PPV were 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 quantification of viral template. The assay was used to measure the levels of virus acquired by several aphid species.

Original languageEnglish (US)
Title of host publicationXIX International Symposium on Virus and Virus-like Diseases of Temperate Fruit Crops - Fruit Tree Diseases
PublisherInternational Society for Horticultural Science
Pages135-139
Number of pages5
ISBN (Print)9789066051485
DOIs
StatePublished - Sep 25 2004

Publication series

NameActa Horticulturae
Volume657
ISSN (Print)0567-7572

All Science Journal Classification (ASJC) codes

  • Horticulture

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    Schneider, W. L., Sherman, D. J., Stone, A. L., Damsteegt, V. D., & Frederick, R. D. (2004). Detection and quantification of Plum pox potyvirus in aphid vectors by real-time fluorescent reverse transcription-PCR. In XIX International Symposium on Virus and Virus-like Diseases of Temperate Fruit Crops - Fruit Tree Diseases (pp. 135-139). (Acta Horticulturae; Vol. 657). International Society for Horticultural Science. https://doi.org/10.17660/ActaHortic.2004.657.17