2025-01-31 ノースカロライナ州立大学(NCState)
<関連情報>
- https://news.ncsu.edu/2025/01/moving-plant-disease-diagnosis-from-lab-to-field-for-faster-results/
- https://apsjournals.apsnet.org/doi/10.1094/PHYTO-02-24-0055-R
マイクロ流体チップとスマートフォンプラットフォームを用いたPhytophthora kernoviae、P. ramorumおよびP. ramorum NA1系統のループ媒介等温増幅検出Loop-Mediated Isothermal Amplification Detection of Phytophthora kernoviae, P. ramorum, and the P. ramorum NA1 Lineage on a Microfluidic Chip and Smartphone Platform
Amanda Mainello-Land, Amanda C. Saville, Jyotsna Acharya, and Jean Ristaino
Phytopathology Published:27 Jan 2025
DOI:https://doi.org/10.1094/PHYTO-02-24-0055-R
Abstract
Rapid, field-deployable assays such as loop-mediated isothermal amplification (LAMP) are critical for detecting nursery and forest pathogens such as Phytophthora ramorum and P. kernoviae to prevent pathogen spread. We developed and validated four LAMP assays for genus-level detection of Phytophthora spp., species-level detection of P. kernoviae and P. ramorum, and lineage-level detection of the P. ramorum NA1 lineage. The cross-reactivity of the two species-specific LAMP assays was evaluated using a set of 18 Phytophthora spp. known to infect nursery crop hosts. The correct target species were detected by the species-level LAMP assays. The Phytophthora spp. LAMP assay was evaluated against 27 Phytophthora spp. and other bacterial and fungal pathogens and reacted with all the Phytophthora spp. evaluated but no other bacterial or fungal species. The limit of detection (LOD) of the P. kernoviae LAMP was 100 fg/µl, and the LOD of the P. ramorum LAMP assay was 1 pg/µl of DNA. The NA1 LAMP assay was tested against the NA1, NA2, EU1, and EU2 lineages of P. ramorum and was lineage-specific but had a higher LOD (100 pg/µl) than the species-specific LAMP assays. Both P. ramorum and P. kernoviae LAMP assays were highly precise (>0.94) in detecting the respective pathogens in symptomatic rhododendron leaves and co-inoculation experiments. The four LAMP assays were run in tandem on a microfluidic chip and smartphone platform and can be used in the field to detect and monitor spread of these regulatory Phytophthora spp. in forest and/or nursery settings.
