Rapid detection of Phytophthora ramorum and P. kernoviae by two-minute DNA extraction followed by isothermal amplification and amplicon detection by generic lateral flow device.
Phytopathology
; 100(2): 143-9, 2010 Feb.
Article
in En
| MEDLINE
| ID: mdl-20055648
ABSTRACT
ABSTRACT A method for nucleic-acid-based detection of pathogens in plant material has been developed which comprises a simple and rapid method for extracting DNA on the nitrocellulose membranes of lateral-flow devices, loop-mediated isothermal amplification (LAMP) of target DNA using labeled primers, and detection of the generically labeled amplification products by a sandwich immunoassay in a lateral-flow-device format. Each of these steps can be performed without specialist equipment and is suitable for on-site use, and a result can be obtained in just over an hour. A LAMP assay for the detection of plant DNA (cytochrome oxidase gene) can be used in conjunction with pathogen-specific assays to confirm negative results. The use of this method is demonstrated for the detection of Phytophthora ramorum, the causal agent of sudden oak death and dieback/leaf blight in a range of tree, shrub, and herbaceous species, and the recently described pathogen P. kernoviae.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Phytophthora
/
Plant Diseases
/
Nucleic Acid Amplification Techniques
/
DNA, Algal
Type of study:
Diagnostic_studies
Language:
En
Journal:
Phytopathology
Journal subject:
BOTANICA
Year:
2010
Type:
Article
Affiliation country:
United kingdom