Faster, simpler, more-specific methods for improved molecular detection of Phytophthora ramorum in the field.
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.
Tomlinson
J A
JA
Central Science Laboratory, Sand Hutton, York YO41 1LZ, United Kingdom. j.tomlinson@csl.gov.uk
Barker
I
I
Boonham
N
N
D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010838:Phytophthora / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D013045:Species Specificity
D017931:DNA Primers