01962nas a2200265 4500008004100000022001400041245015100055210006900206260002500275300001200300490000800312520109300320653002701413653002701440653001701467653001701484100002301501700002301524700002401547700002101571700001801592700001801610700002001628856004801648 2011 eng d a0929-187300aDevelopment of a real-time PCR assay for detection of Phytophthora kernoviae and comparison of this method with a conventional culturing technique0 aDevelopment of a realtime PCR assay for detection of Phytophthor bSpringer Netherlands a695-7030 v1313 a

Phytophthora kernoviae is a recently described pathogen causing leaf blight, aerial dieback and bleeding cankers on trees and shrubs in parts of Great Britain and Ireland and recently reported in New Zealand. This paper describes the development of a TaqMan real-time PCR assay based on internal transcribed spacer (ITS) sequence to aid diagnosis of this pathogen in culture and in plant material. The assay showed no cross reaction with 29 other Phytophthora species, including the closely related species P. boehmeriae, and detected at least 1.2 pg of P. kernoviae DNA per reaction. A rapid and simple method can be used to extract DNA prior to testing by real-time PCR, and a plant internal control assay can be used to aid interpretation of negative results. A comparison of real-time PCR and plating for 526 plant samples collected in the UK indicated that this assay is suitable for use in routine screening for P. kernoviae.

10aDiagnostic sensitivity10aDiagnostic specificity10aplant health10aRhododendron1 aHughes, KelvinJ.D.1 aTomlinson, JennyA.1 aGiltrap, PatriciaM.1 aBarton, Victoria1 aHobden, Ellie1 aBoonham, Neil1 aLane, CharlesR. uhttp://dx.doi.org/10.1007/s10658-011-9843-x