ArticleScientific reports2016
Direct identification of antibiotic resistance genes on single plasmid molecules using CRISPR/Cas9 in combination with optical DNA mapping.
Article in Scientific reports, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 45 papers.
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Who cites it
45 citing papers in PubMed.
- CRISPR/Cas system as a novel therapeutic strategy to combat multi-drug-resistant bacteria.Archives of microbiology · 2026Review
- Programmable CRISPR-Cas diagnostic platforms for rapid detection of uropathogens and antimicrobial resistance.Archives of microbiology · 2026Review
- Cas9-Leveraged Single-Molecule Characterization of Sparse Plasmid Vectors in Heterogenous DNA Samples.Applied biochemistry and biotechnology · 2026Article
- Emergence of carbapenem-producing enterobacteriaceae (CPE) and other multidrug-resistant gram-negative bacteria in neonates at a tertiary-level NICU in Tanzania: a point prevalence study.JAC-antimicrobial resistance · 2025Article
- Detection of antimicrobial resistance via state-of-the-art technologies versus conventional methods.Frontiers in microbiology · 2025Review
- Detecting antibiotic resistance: classical, molecular, advanced bioengineering, and AI-enhanced approaches.Frontiers in microbiology · 2025Review
- Portable microfluidic devices for monitoring antibiotic resistance genes in wastewater.Mikrochimica acta · 2024Review
- Mitigating Antibiotic Resistance: The Utilization of CRISPR Technology in Detection.Biosensors · 2024Review
- Comparison of CTX-M encoding plasmids present during the early phase of the ESBL pandemic in western Sweden.Scientific reports · 2024Article
- Recent Advances in Understanding the Molecular Mechanisms of Multidrug Resistance and Novel Approaches of CRISPR/Cas9-Based Genome-Editing to Combat This Health Emergency.International journal of nanomedicine · 2024Review
- CRISPR/Cas9 Landscape: Current State and Future Perspectives.International journal of molecular sciences · 2023Review
- Strain-level bacterial typing directly from patient samples using optical DNA mapping.Communications medicine · 2023Article
- Identification and characterization of plasmids carrying the mobile colistin resistance geneJAC-antimicrobial resistance · 2023Article
- CRISPR-cas technology: A key approach for SARS-CoV-2 detection.Frontiers in bioengineering and biotechnology · 2023Review
- Recent Advances in CRISPR-Cas Technologies for Synthetic Biology.Journal of microbiology (Seoul, Korea) · 2023Review
- A simple cut and stretch assay to detect antimicrobial resistance genes on bacterial plasmids by single-molecule fluorescence microscopy.Scientific reports · 2022Article
- Article
- Extended-Spectrum Beta-Lactamases ProducingInfection and drug resistance · 2022Review
- Review
- Engineered CRISPR-Cas systems for the detection and control of antibiotic-resistant infections.Journal of nanobiotechnology · 2021Review
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Authors and funding
10 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Bacterial plasmids are extensively involved in the rapid global spread of antibiotic resistance. We here present an assay, based on optical DNA mapping of single plasmids in nanofluidic channels, which provides detailed information about the plasmids present in a bacterial isolate. In a single experiment, we obtain the number of different plasmids in the sample, the size of each plasmid, an optical barcode that can be used to identify and trace the plasmid of interest and information about which plasmid that carries a specific resistance gene. Gene identification is done using CRISPR/Cas9 loaded with a guide-RNA (gRNA) complementary to the gene of interest that linearizes the circular plasmids at a specific location that is identified using the optical DNA maps. We demonstrate the principle on clinically relevant extended spectrum beta-lactamase (ESBL) producing isolates. We discuss how the gRNA sequence can be varied to obtain the desired information. The gRNA can either be very specific to identify a homogeneous group of genes or general to detect several groups of genes at the same time. Finally, we demonstrate an example where we use a combination of two gRNA sequences to identify carbapenemase-encoding genes in two previously not characterized clinical bacterial samples.
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