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

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...

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Bibliographic Details
Published inActa horticulturae no. 657; pp. 135 - 139
Main Authors Schneider, W.L, Sherman, D.J, Stone, A.L, Damsteegt, V.D, Frederick, R.D
Format Journal Article
LanguageEnglish
Published 01.01.2004
Subjects
Acyrthosiphon pisum
Aphis spiraecola
Brachycaudus
Brachycaudus persicae
fluorescence
insect vectors
microbial detection
Myzus persicae
Plum pox virus
Prunus
reverse transcriptase polymerase chain reaction
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Summary: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.
Bibliography:http://hdl.handle.net/10113/1030
ISSN:0567-7572
DOI:10.17660/ActaHortic.2004.657.17