TitleOn-Site Molecular Detection of Soil-Borne Phytopathogens Using a Portable Real-Time PCR System.
Publication TypeJournal Article
Year of Publication2018
AuthorsDeShields, JB, Bomberger, RA, Woodhall, JW, Wheeler, DL, Moroz, N, Johnson, DA, Tanaka, K
JournalJ Vis Exp
Issue132
Date Published2018 02 23
ISSN1940-087X
KeywordsPlant Pathology, Real-Time Polymerase Chain Reaction, Soil
Abstract

On-site diagnosis of plant diseases can be a useful tool for growers for timely decisions enabling the earlier implementation of disease management strategies that reduce the impact of the disease. Presently in many diagnostic laboratories, the polymerase chain reaction (PCR), particularly real-time PCR, is considered the most sensitive and accurate method for plant pathogen detection. However, laboratory-based PCRs typically require expensive laboratory equipment and skilled personnel. In this study, soil-borne pathogens of potato are used to demonstrate the potential for on-site molecular detection. This was achieved using a rapid and simple protocol comprising of magnetic bead-based nucleic acid extraction, portable real-time PCR (fluorogenic probe-based assay). The portable real-time PCR approach compared favorably with a laboratory-based system, detecting as few as 100 copies of DNA from Spongospora subterranea. The portable real-time PCR method developed here can serve as an alternative to laboratory-based approaches and a useful on-site tool for pathogen diagnosis.

DOI10.3791/56891
Alternate JournalJ Vis Exp
PubMed ID29553557
PubMed Central IDPMC5931365