Robust design of LAMP assays for in-field detection of major bacterial vascular diseases of banana

作者：Isabelle Robeǹe, Mikel Arrieta-Salgado, Samuel Rozsasi, Diane Mostert, Véronique Maillot-Lebon, Yolande Chilin-Charles, Jos Jansen van Vuuren, Gloria Valentine Nakato, Reagan Kanaabi, Freddy Magdama, E. Salgado, Catur Hermanto, Agus Sutanto, Janet Conie, Debbie Henry, Sheryl Le Roux, Altus Viljoen, Emmanuel Jouen, Mirana Gauche, Stéphane Poussier, Emmanuel Wicker, Yann Pécrix · 发表于：PLoS ONE · 年份：2026 · DOI：10.1371/journal.pone.0337387 · 研究领域：Biosensors and Analytical Detection、Plant Pathogenic Bacteria Studies、Vibrio bacteria research studies

Bacterial diseases of banana are a growing global threat, causing yield losses and increased management costs. Major diseases include Moko, banana blood disease (BBD), and banana Xanthomonas wilt (BXW), caused by Ralstonia solanacearum, Ralstonia syzygii subsp. celebesensis, and Xanthomonas vasicola pv. musacearum, respectively. Effective surveillance requires point-of-care diagnostics such as loop-mediated isothermal amplification (LAMP) for on-site use. We aimed to develop three LAMP assays to specifically detect the bacteria responsible for Moko, BBD and BXW, directly from banana tissues, using a simplified DNA extraction protocol. The BBD - and BXW-LAMP assays demonstrated 100% specificity, yielding negative results for a broad range of non-target bacteria, including closely related species as well as pathogenic and endophytic strains associated with banana, and positive results for all the tested target strains. For Moko disease, a duplex-LAMP assay was developed to detect all strains from the four globally most relevant sequevars: IIB-3, IIB-4, IIA-6, and IIA-24. The duplex-LAMP successfully detected all target strains, except one that was shown to be non-pathogenic to Cavendish bananas. All non-target strains tested negative, with the exception of a delayed signal for one strain belonging to Ralstonia thomasi, not associated with banana environment (hospital strain). These results were supported by an extensive in silico analysis conducted on 9,668 Burkholderiaceae and...