Faster, Simpler, More-Specific Methods for Improved Molecular Detection of Phytophthora ramorum in the Field

• Published in: Applied and Environmental Microbiology Vol. 73; no. 12; pp. 4040 - 4047
• Main Authors: Tomlinson, J.A, Barker, I, Boonham, N
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01.06.2007
• Subjects: Base Sequence; Biological and medical sciences; DNA Primers - genetics; Fundamental and applied biological sciences. Psychology; Methods; Microbiology; Molecular Sequence Data; Nucleic Acid Amplification Techniques - methods; Pathogens; Phytophthora; Phytophthora - genetics; Polymerase Chain Reaction - methods; Real time; Species Specificity; Trees; Fungi; Method; Phytophthora; Detection; Phycomycetes; Thallophyta
• ISSN: 0099-2240; 1098-5336
• DOI: 10.1128/AEM.00161-07

Phytophthora ramorum is the causal agent of sudden oak death. The pathogen also affects a wide range of tree, shrub, and herbaceous species in natural and landscaped environments as well as plants in the nursery industry. A TaqMan real-time PCR method for the detection of this pathogen in the field has been described previously; this paper describes the development of a number of assays based on this method which have various advantages for use in the field. A scorpion real-time PCR assay that is twice as fast as TaqMan was developed, allowing the detection of P. ramorum in less than 30 min. Also designed was a loop-mediated isothermal amplification (LAMP) assay, which allowed sensitive and specific detection of P. ramorum in 45 min using only a heated block. A positive reaction was identified by the detection of the LAMP product by color change visible to the naked eye.