A Simplified, CRISPR-Based Method for the Detection of Batrachochytrium salamandrivorans.

Autor: Hoenig BD; Department of Biological Sciences, University of Pittsburgh, 4200 Fifth Ave, Pittsburgh, PA, 15260, USA. brandonhoenig@gmail.com.; Pymatuning Laboratory of Ecology, Linesville, PA, USA. brandonhoenig@gmail.com., Böning P; Biogeography Department, Trier University, Universitätsring 15, 54296, Trier, Germany., Plewnia A; Biogeography Department, Trier University, Universitätsring 15, 54296, Trier, Germany., Richards-Zawacki CL; Department of Biological Sciences, University of Pittsburgh, 4200 Fifth Ave, Pittsburgh, PA, 15260, USA.; Pymatuning Laboratory of Ecology, Linesville, PA, USA.
Jazyk: angličtina
Zdroj: EcoHealth [Ecohealth] 2024 Nov 29. Date of Electronic Publication: 2024 Nov 29.
DOI: 10.1007/s10393-024-01690-x
Abstrakt: The fungal pathogen Batrachochytrium salamandrivorans (Bsal) is one of two species (the other, B. dendrobatidis/Bd) that cause amphibian chytridiomycosis, an emerging infectious disease that has been indicated in the declines of hundreds of amphibian species worldwide. While Bd has been near globally distributed for decades, Bsal is a more recently emerged pathogen, having been identified just over a decade ago with current impacts localized to salamandrids in parts of Europe. However, because there is concern that Bsal will cause widespread declines if introduced to naïve regions-such as the Americas where the greatest diversity of salamandrids exist-it is imperative that widespread monitoring strategies be implemented to mitigate the spread of Bsal. As standard molecular diagnostic approaches-such as qPCR-tend to be expensive, time-consuming, or require specialized instrumentation and training, we have developed a simplified, rapid, CRISPR-based approach for Bsal-DNA detection. Here, we demonstrate this approach-termed FINDeM (Field-deployable, Isothermal, Nucleotide-based Detection Method)-and show that it can detect clinically relevant concentrations of Bsal DNA in under an hour using only inexpensive supplies and body-heat inducible reactions. Further, we highlight drawbacks and limitations associated with FINDeM-such as decreased DNA extraction yields and detection sensitivity when compared to more traditional approaches-and provide suggestions for additional development and future application of this method.
Competing Interests: Declarations. Conflict of interest: The authors declare no conflict of interest.
(© 2024. EcoHealth Alliance.)
Databáze: MEDLINE
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