Development of a recombinase polymerase amplification method combined with a lateral flow dipstick assay for rapid detection of the larch pathogen Neofusicoccum laricinum
Neofusicoccum laricinum, an important pathogenic species, causes shoot blight of larch. In China, large areas of Larix principis-rupprechtii forests are threatened by this pathogen. Currently, this pathogen is on the list of quarantine pests in Chinese. Due to the widespread and severe damage caused...
| Veröffentlicht in: | Plant disease. - 1997. - (2024) vom: 23. Sept. |
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| 1. Verfasser: | |
| Weitere Verfasser: | , , , |
| Format: | Online-Aufsatz |
| Sprache: | English |
| Veröffentlicht: |
2024
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| Zugriff auf das übergeordnete Werk: | Plant disease |
| Schlagworte: | Journal Article Causal Agent Crop Type Fungi Pathogen detection Subject Areas Techniques Trees forest |
| Zusammenfassung: | Neofusicoccum laricinum, an important pathogenic species, causes shoot blight of larch. In China, large areas of Larix principis-rupprechtii forests are threatened by this pathogen. Currently, this pathogen is on the list of quarantine pests in Chinese. Due to the widespread and severe damage caused by N. laricinum, a reliable and accurate diagnostic tool is urgently needed. In this study, we first identified a Nlar12009 as a N. laricinum-specific gene through genomic sequence data and bioinformatic analysis. Specific primer pairs and DNA probes were designed to detect the target pathogen using a novel recombinase polymerase amplification assay with a lateral flow dipstick (RPA-LFD) method. We optimized the RPA-LFD assay to ensure high specificity to N. laricinum. Our results showed that the assay exclusively detected N. laricinium isolates with no cross-reaction with other isolates of fungaland oomycete species and nematodes. Furthermore, our detection technique exhibited a 10-fold higher sensitivity (10 fg/mL) than conventional polymerase chain reaction (PCR) for N. laricinum detection. Our developed RPA-LFD assay is proved to be a highly specific, sensitive, time-saving, and convenient method for the diagnosis of N. laricinum and shows great potential in field application |
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| Beschreibung: | Date Revised 24.09.2024 published: Print-Electronic Citation Status Publisher |
| ISSN: | 0191-2917 |
| DOI: | 10.1094/PDIS-05-24-1033-SR |