CRISPR-based diagnostics detects invasive insect pests.

Autor:	Shashank PR; McGuire Center for Lepidoptera and Biodiversity, Florida Museum of Natural History, University of Florida, Gainesville, FL 32611, USA.; Division of Entomology, ICAR-Indian Agricultural Research Institution, New Delhi 110012, India., Parker BM; McGuire Center for Lepidoptera and Biodiversity, Florida Museum of Natural History, University of Florida, Gainesville, FL 32611, USA.; Oak Ridge Institute for Science and Education, Oak Ridge, TN 37831, USA.; U.S. Environmental Protection Agency, Office of Research and Development, RTP, NC, 27709, USA., Rananaware SR; Department of Chemical Engineering, University of Florida, Gainesville, FL 32611, USA., Plotkin D; McGuire Center for Lepidoptera and Biodiversity, Florida Museum of Natural History, University of Florida, Gainesville, FL 32611, USA., Couch C; McGuire Center for Lepidoptera and Biodiversity, Florida Museum of Natural History, University of Florida, Gainesville, FL 32611, USA., Yang LG; Department of Chemical Engineering, University of Florida, Gainesville, FL 32611, USA., Nguyen LT; Department of Chemical Engineering, University of Florida, Gainesville, FL 32611, USA., Prasannakumar NR; Division of Crop Protection, ICAR-Indian Institute of Horticultural Research, Bengaluru 560089, India., Braswell WE; Insect Management and Molecular Diagnostics Laboratory, USDA APHIS PPQ S&T, 22675 North Moorefield Road, Edinburg, Texas 78541, USA., Jain PK; Department of Chemical Engineering, University of Florida, Gainesville, FL 32611, USA.; Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, Florida, USA.; UF Health Cancer Center, University of Florida, Gainesville, Florida, USA., Kawahara AY; McGuire Center for Lepidoptera and Biodiversity, Florida Museum of Natural History, University of Florida, Gainesville, FL 32611, USA.
Jazyk:	angličtina
Zdroj:	BioRxiv : the preprint server for biology [bioRxiv] 2023 May 18. Date of Electronic Publication: 2023 May 18.
DOI:	10.1101/2023.05.16.541004
Abstrakt:	Rapid identification of organisms is essential across many biological and medical disciplines, from understanding basic ecosystem processes and how organisms respond to environmental change, to disease diagnosis and detection of invasive pests. CRISPR-based diagnostics offers a novel and rapid alternative to other identification methods and can revolutionize our ability to detect organisms with high accuracy. Here we describe a CRISPR-based diagnostic developed with the universal cytochrome-oxidase 1 gene (CO1). The CO1 gene is the most sequenced gene among Animalia, and therefore our approach can be adopted to detect nearly any animal. We tested the approach on three difficult-to-identify moth species ( Keiferia lycopersicella, Phthorimaea absoluta , and Scrobipalpa atriplicella ) that are major invasive pests globally. We designed an assay that combines recombinase polymerase amplification (RPA) with CRISPR for signal generation. Our approach has a much higher sensitivity than other real time-PCR assays and achieved 100% accuracy for identification of all three species, with a detection limit of up to 120 fM for P. absoluta and 400 fM for the other two species. Our approach does not require a lab setting, reduces the risk of cross-contamination, and can be completed in less than one hour. This work serves as a proof of concept that has the potential to revolutionize animal detection and monitoring. Competing Interests: Competing Interest Statement: The authors declare no competing interests.
Databáze:	MEDLINE
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