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United States Department of Agriculture

Agricultural Research Service

Title: Specific Detection and Quantification of Plum Pox Potyvirus by Real-Time Fluorescent Rt-Pcr

Authors
item Schneider, William
item Sherman, Diana
item Stone, Andrew
item Damsteegt, Vernon
item Frederick, Reid

Submitted to: American Phytopathological Society Abstracts
Publication Type: Abstract Only
Publication Acceptance Date: March 31, 2002
Publication Date: July 1, 2002

Technical Abstract: Plum pox potyvirus (PPV), economically the most important stone fruit virus in the world, was identified in Adams County, PA in 1999. Surveys continue in the surrounding areas for detection and eradication of trees infected with PPV. The survey uses a PPV monoclonal antibody ELISA-DASI assay, which is time consuming and somewhat unreliable in detecting low titer viral infections. A real-time fluorescent RT-PCR assay was developed to detect and quantify PPV in tissue samples using multiple platforms (i.e. Smart Cycler, TaqMan, etc.). The target sequence was selected in a conserved region of the coat protein. Detection limits (750 fg of target RNA) and standard curves were determined using in vitro transcripts that included the target sequence. The assay successfully identified European and Pennsylvania strains of PPV in leaf tissue samples from plum, peach, pea, N. clevelandii and N. benthamiana. Ongoing work includes adapting the assay to index different Prunus tissues (i.e. roots, stems, bark, buds and flowers).

Last Modified: 12/19/2014
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