01988nas a2200193   4500008004100000022001400041245010400055210006900159260001600228300001600244490000700260520139300267100001901660700001601679700001801695700002001713700001501733856004601748       2005                        eng d  a0099-224000aOn-Site DNA Extraction and Real-Time PCR for Detection of Phytophthora ramorum in the FieldABSTRACT0 aOnSite DNA Extraction and RealTime PCR for Detection of Phytopht  cJan-11-2005  a6702 - 67100 v713 a<p>Phytophthora ramorum is a recently described pathogen causing oak mortality (sudden oak death) in forests in coastal areas of California and southern Oregon and dieback and leaf blight in a range of tree, shrub, and herbaceous species in the United States and Europe. Due to the threat posed by this organism, stringent quarantine regulations are in place, which restrict the movement of a number of hosts. Fast and accurate diagnostic tests are required in order to characterize the distribution of P. ramorum, prevent its introduction into pathogen-free areas, and minimize its spread within affected areas. However, sending samples to a laboratory for testing can cause a substantial delay between sampling and diagnosis. A rapid and simple DNA extraction method was developed for use at the point of sampling and used to extract DNAs from symptomatic foliage and stems in the field. A sensitive and specific single-round real-time PCR (TaqMan) assay for P. ramorum was performed using a portable real-time PCR platform (Cepheid SmartCycler II), and a cost-effective method for stabilizing PCR reagents was developed to allow their storage and transportation at room temperature. To our knowledge, this is the first description of a method for DNA extraction and molecular testing for a plant pathogen carried out entirely in the field, independent of any laboratory facilities.</p>1 aTomlinson, J A1 aBoonham, N.1 aHughes, K J D1 aGriffin, R., L.1 aBarker, I.  uhttps://pubmed.ncbi.nlm.nih.gov/16269700/