A five-minute DNA extraction method for expedited detection of Phytophthora ramorum following prescreening using Phytophthora spp. lateral flow devices.
In a direct comparison with established methods for Phytophthora ramorum detection (isolation followed by morphological identification, or conventional DNA extraction followed by TaqMan real-time PCR) a rapid, simplified detection method in which membranes of lateral flow devices (LFDs) are added directly to TaqMan real-time PCR reactions was used to test 202 plant samples collected by plant health inspectors in the field. P. ramorum prevalence within the 202 samples was approximately 40% according to routine testing by isolation or TaqMan real-time PCR. The diagnostic sensitivity and specificity of the rapid detection method were 96.3% and 91.2%, respectively. This method can be used in conjunction with Phytophthora spp. lateral flow devices to reduce the number of samples requiring testing using more laborious conventional methods. The effect of combining prescreening for Phytophthora spp. with P. ramorum-specific tests is discussed in terms of the positive and negative predictive values of species-specific detection when testing samples collected in different inspection scenarios.
Tomlinson
J A
JA
The Food and Environment Research Agency, Sand Hutton, York YO41 1LZ, United Kingdom. jenny.tomlinson@fera.gsi.gov.uk
Dickinson
M
M
Hobden
E
E
Robinson
S
S
Giltrap
P M
PM
Boonham
N
N
D004247:DNA / Q000302:isolation & purification*; D005374:Filtration / Q000379:methods; D008828:Microbiological Techniques / Q000379:methods*; D010838:Phytophthora / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013997:Time Factors
D004247:DNA