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 Citations (Scopus)

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

Fingerprint

Plum pox virus
Potyvirus
Aphidoidea
reverse transcriptase polymerase chain reaction
assays
Prunus
buds
enzyme-linked immunosorbent assay
viruses
stems
leaves

All Science Journal Classification (ASJC) codes

  • Horticulture

Cite this

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
Schneider, William L. ; Sherman, Diana J. ; Stone, Andrew L. ; Damsteegt, Vernon D. ; Frederick, Reid David. / Detection and quantification of Plum pox potyvirus in aphid vectors by real-time fluorescent reverse transcription-PCR. XIX International Symposium on Virus and Virus-like Diseases of Temperate Fruit Crops - Fruit Tree Diseases. International Society for Horticultural Science, 2004. pp. 135-139 (Acta Horticulturae).
@inproceedings{ddaa81df4d4844209a2a10dd65838975,
title = "Detection and quantification of Plum pox potyvirus in aphid vectors by real-time fluorescent reverse transcription-PCR",
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.",
author = "Schneider, {William L.} and Sherman, {Diana J.} and Stone, {Andrew L.} and Damsteegt, {Vernon D.} and Frederick, {Reid David}",
year = "2004",
month = "9",
day = "25",
doi = "10.17660/ActaHortic.2004.657.17",
language = "English (US)",
isbn = "9789066051485",
series = "Acta Horticulturae",
publisher = "International Society for Horticultural Science",
pages = "135--139",
booktitle = "XIX International Symposium on Virus and Virus-like Diseases of Temperate Fruit Crops - Fruit Tree Diseases",
address = "Belgium",

}

Schneider, WL, Sherman, DJ, Stone, AL, Damsteegt, VD & Frederick, RD 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. Acta Horticulturae, vol. 657, International Society for Horticultural Science, pp. 135-139. https://doi.org/10.17660/ActaHortic.2004.657.17

Detection and quantification of Plum pox potyvirus in aphid vectors by real-time fluorescent reverse transcription-PCR. / Schneider, William L.; Sherman, Diana J.; Stone, Andrew L.; Damsteegt, Vernon D.; Frederick, Reid David.

XIX International Symposium on Virus and Virus-like Diseases of Temperate Fruit Crops - Fruit Tree Diseases. International Society for Horticultural Science, 2004. p. 135-139 (Acta Horticulturae; Vol. 657).

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

TY - GEN

T1 - Detection and quantification of Plum pox potyvirus in aphid vectors by real-time fluorescent reverse transcription-PCR

AU - Schneider, William L.

AU - Sherman, Diana J.

AU - Stone, Andrew L.

AU - Damsteegt, Vernon D.

AU - Frederick, Reid David

PY - 2004/9/25

Y1 - 2004/9/25

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=34548148778&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34548148778&partnerID=8YFLogxK

U2 - 10.17660/ActaHortic.2004.657.17

DO - 10.17660/ActaHortic.2004.657.17

M3 - Conference contribution

AN - SCOPUS:34548148778

SN - 9789066051485

T3 - Acta Horticulturae

SP - 135

EP - 139

BT - XIX International Symposium on Virus and Virus-like Diseases of Temperate Fruit Crops - Fruit Tree Diseases

PB - International Society for Horticultural Science

ER -

Schneider WL, Sherman DJ, Stone AL, Damsteegt VD, Frederick RD. 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. International Society for Horticultural Science. 2004. p. 135-139. (Acta Horticulturae). https://doi.org/10.17660/ActaHortic.2004.657.17