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Applied and Environmental Microbiology, June 2007, p. 4040-4047, Vol. 73, No. 12
0099-2240/07/$08.00+0     doi:10.1128/AEM.00161-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

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

J. A. Tomlinson,* I. Barker, and N. Boonham

Central Science Laboratory, Sand Hutton, York YO41 1LZ, United Kingdom

Received 22 January 2007/ Accepted 13 April 2007

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.

* Corresponding author. Mailing address: Central Science Laboratory, Sand Hutton, York YO41 1LZ, United Kingdom. Phone: (44) 1904 462000, ext. 3215. Fax: (44) 1904 462111. E-mail: j.tomlinson{at}csl.gov.uk

{triangledown} Published ahead of print on 20 April 2007.

Applied and Environmental Microbiology, June 2007, p. 4040-4047, Vol. 73, No. 12
0099-2240/07/$08.00+0     doi:10.1128/AEM.00161-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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