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Development of a Sensitive Colorimetric-PCR Assay for Detection of Viruses in Woody Plants

August 1998 , Volume 82 , Number  8
Pages  880 - 884

Adib Rowhani , Plant Pathology Specialist , Lenka Biardi , Postgraduate Researcher , Geoffrey Routh , Postdoctoral Fellow , Steve D. Daubert , Staff Research Associate , and Deborah A. Golino , Extension Specialist, Department of Plant Pathology, University of California, Davis 95616

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Accepted for publication 17 April 1998.

Diagnostic methods employing the polymerase chain reaction (PCR) provide the most sensitive means currently available for detecting viruses in woody plants. A new technique has been tested that does not rely on gel electrophoresis or molecular hybridization to detect virus-specific PCR products. This colorimetric method for detection of PCR products from woody plants was demonstrated to be at least as sensitive as gel analysis. When combined with immunocapture of virions from plant sap, colorimetric detection provides a means to apply PCR technology to a large number of samples. Here, we report on the use of this technique for detection and quantitation of a walnut isolate of cherry leafroll virus (CLRV-W), citrus tristeza virus (CTV), prune dwarf virus (PDV), prunus necrotic ringspot virus (PNRSV), and tomato ringspot virus (ToRSV) in woody and herbaceous plants. For purified virus preparations, detection limits ranged from 100 pg/ml to 100 ag/ml. We also describe the colorimetric PCR detection of CTV in pooled samples.

© 1998 The American Phytopathological Society