Protocols

To determine if living microorganisms of phytosanitary concern are present in wood after eradication treatment and to evaluate the efficacy of such treatments, the method of choice is to grow microbes in petri dishes for subsequent identification. However, some plant pathogens are difficult or impossible to grow in axenic cultures. A molecular methodology capable of detecting living fungi and fungus-like organisms in situ can provide a solution.

Reference publication: Wong, B.,  Leal, I., Feau, N., Dale, A.,  Uzunovic, A. and Hamelin, R. C. 2019. Molecular assays to detect the presence and viability of Phytophthora ramorum and Grosmannia clavigera. bioRxiv 736637; doi: https://doi.org/10.1101/736637

Molecular diagnostics by DNA testing for environmental applications requires a cost-effective and efficient point-of-use DNA test that can obtain accurate results from remote sites in real-time. This requires the development of simple and fast sample processing and DNA extraction, room-temperature stable reagents and a portable instrument. We developed a point-of-use real-time Polymerase Chain Reaction system using a crude buffer-based DNA extraction protocol and lyophilized, pre-made, reactions for on-site applications. Here is the standard operating protocol for assembling the kit described. 

Reference publication: This standard operating protocol is based on the following publication: Capron A, Stewart D, Hrywkiw K, Allen K, Feau N, Bilodeau G, et al. (2020) In Situ Processing and Efficient Environmental Detection (iSPEED) of tree pests and pathogens using point-of-use real-time PCR. PLoS ONE 15(4): e0226863. https://doi.org/10.1371/journal.pone.0226863

To determine if living microorganisms of phytosanitary concern are present in wood after eradication treatment and to evaluate the efficacy of such treatments, the method of choice is to grow microbes in petri dishes for subsequent identification. However, some plant pathogens are difficult or impossible to grow in axenic cultures. A molecular methodology capable of detecting living fungi and fungus-like organisms in situ can provide a solution.

Reference publication: Wong, B.,  Leal, I., Feau, N., Dale, A.,  Uzunovic, A. and Hamelin, R. C. 2019. Molecular assays to detect the presence and viability of Phytophthora ramorum and Grosmannia clavigera. bioRxiv 736637; doi: https://doi.org/10.1101/736637

Molecular diagnostics by DNA testing for environmental applications requires a cost-effective and efficient point-of-use DNA test that can obtain accurate results from remote sites in real-time. This requires the development of simple and fast sample processing and DNA extraction, room-temperature stable reagents and a portable instrument. We developed a point-of-use real-time Polymerase Chain Reaction system using a crude buffer-based DNA extraction protocol and lyophilized, pre-made, reactions for on-site applications. Here is the standard operating protocol for assembling the kit described. 

Reference publication: This standard operating protocol is based on the following publication: Capron A, Stewart D, Hrywkiw K, Allen K, Feau N, Bilodeau G, et al. (2020) In Situ Processing and Efficient Environmental Detection (iSPEED) of tree pests and pathogens using point-of-use real-time PCR. PLoS ONE 15(4): e0226863. https://doi.org/10.1371/journal.pone.0226863

Manual for a portable assay to detect Asian Spongy Moth (ASM) from egg mass samples. This system consists of a single triplex reaction (“Triplex 1”) targeting three different mitochondrial marker genes. It will identify moths from the ASM complex (L. dispar asiatica, L. dispar japonica and L. umbrosa).