Faster, simpler, more-specific methods for improved molecular detection of Phytophthora ramorum in the field

Appl Environ Microbiol. 2007 Jun;73(12):4040-7. doi: 10.1128/AEM.00161-07. Epub 2007 Apr 20.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA Primers / genetics
  • Molecular Sequence Data
  • Nucleic Acid Amplification Techniques / methods*
  • Phytophthora / genetics*
  • Polymerase Chain Reaction / methods*
  • Species Specificity

Substances

  • DNA Primers