Canadian Forest Service Publications
Development of a Rapid Loop-Mediated Isothermal Amplification assay for the detection of Dothistroma septosporum. 2021. Myrholm, C.L.; Tomm, B.D.; Heinzelmann, R.; Feau, N.; Hamelin, R.C.; McDougal, R.; Winkworth, R.C.; Ramsfield, T.D. Forests 12(3):362.
Year: 2021
Issued by: Northern Forestry Centre
Catalog ID: 40704
Language: English
Availability: PDF (download)
Available from the Journal's Web site. †
DOI: 10.3390/f12030362
† This site may require a fee
Abstract
A Loop-Mediated Isothermal Amplification (LAMP) assay was developed for the detection of the pine pathogen Dothistroma septosporum (G. Dorog.) M. Morelet. The specificity of the LAMP assay was tested using a selection of pine needle fungi, including Dothistroma pini Hulbary, and Lecanosticta acicola (Thüm.) Syd.; only D. septosporum DNA was amplified by the test. In terms of sensitivity, the assay was able to detect as little as 1 pg of total D. septosporum DNA. This assay enables DNA extracted from diseased host needles to be rapidly tested for the presence of D. septosporum using relatively simple to operate equipment away from a fully equipped molecular biology laboratory.
Plain Language Summary
This manuscript outlines the development and testing of a DNA-based diagnostic technique for Dothistroma septosporum, a foliar pathogen of pines. The assay can be conducted in the field using the GENIE-III instrument, which allows rapid and sensitive detection of the pathogen within infected tissue. The assay was developed in order to allow us to rapidly identify the pathogen under field conditions. As this pathogen is globally important, it is hoped that the assay will be useful in other countries where the pathogen is present.
