title: High-throughput screening of antibiotic-resistant bacteria in picodroplets.
abstract: The prevalence of clinically-relevant bacterial strains resistant to current antibiotic therapies is increasing and has been recognized as a major health threat. For example, multidrug-resistant tuberculosis and methicillin-resistant Staphylococcus aureus are of global concern. Novel methodologies are needed to identify new targets or novel compounds unaffected by pre-existing resistance mechanisms. Recently, water-in-oil picodroplets have been used as an alternative to conventional high-throughput methods, especially for phenotypic screening. Here we demonstrate a novel microfluidic-based picodroplet platform which enables high-throughput assessment and isolation of antibiotic-resistant bacteria in a label-free manner. As a proof-of-concept, the system was used to isolate fusidic acid-resistant mutants and estimate the frequency of resistance among a population of Escherichia coli (strain HS151). This approach can be used for rapid screening of rare antibiotic-resistant mutants to help identify novel compound/target pairs.
authors.affiliation: Sphere Fluidics Limited, The Jonas Webb Building, Babraham Research Campus, Babraham, Cambridge, CB22 3AT, UK. xin.liu@spherefluidics.com clive.smith@spherefluidics.com.
authors.forename: X
authors.initials: X
authors.lastname: Liu
authors.affiliation: Merck Research Labs, Merck & Co., Inc., 2015 Galloping Hill Road, K15, Kenilworth, NJ07033, USA.
authors.forename: R E
authors.initials: RE
authors.lastname: Painter
authors.affiliation: Sphere Fluidics Limited, The Jonas Webb Building, Babraham Research Campus, Babraham, Cambridge, CB22 3AT, UK. xin.liu@spherefluidics.com clive.smith@spherefluidics.com.
authors.forename: K
authors.initials: K
authors.lastname: Enesa
authors.affiliation: Sphere Fluidics Limited, The Jonas Webb Building, Babraham Research Campus, Babraham, Cambridge, CB22 3AT, UK. xin.liu@spherefluidics.com clive.smith@spherefluidics.com.
authors.forename: D
authors.initials: D
authors.lastname: Holmes
authors.affiliation: School of Engineering & Physical Sciences, Institute of Biological Chemistry, Biophysics & Bioengineering, Heriot-Watt University, Edinburgh, EH14 4AS, UK.
authors.forename: G
authors.initials: G
authors.lastname: Whyte
authors.affiliation: Merck Research Labs, Merck & Co., Inc., 2015 Galloping Hill Road, K15, Kenilworth, NJ07033, USA.
authors.forename: C G
authors.initials: CG
authors.lastname: Garlisi
authors.affiliation: Merck Research Labs, Merck & Co., Inc., 2015 Galloping Hill Road, K15, Kenilworth, NJ07033, USA.
authors.forename: F J
authors.initials: FJ
authors.lastname: Monsma
authors.affiliation: Sphere Fluidics Limited, The Jonas Webb Building, Babraham Research Campus, Babraham, Cambridge, CB22 3AT, UK. xin.liu@spherefluidics.com clive.smith@spherefluidics.com.
authors.forename: M
authors.initials: M
authors.lastname: Rehak
authors.affiliation: Sphere Fluidics Limited, The Jonas Webb Building, Babraham Research Campus, Babraham, Cambridge, CB22 3AT, UK. xin.liu@spherefluidics.com clive.smith@spherefluidics.com.
authors.forename: F F
authors.initials: FF
authors.lastname: Craig
authors.affiliation: Sphere Fluidics Limited, The Jonas Webb Building, Babraham Research Campus, Babraham, Cambridge, CB22 3AT, UK. xin.liu@spherefluidics.com clive.smith@spherefluidics.com.
authors.forename: C A
authors.initials: CA
authors.lastname: Smith
mesh_terms: D000465:Algorithms; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D018914:Cells, Immobilized; D024881:Drug Resistance, Bacterial; D024901:Drug Resistance, Multiple, Bacterial; D004655:Emulsions; D004867:Equipment Design; D004926:Escherichia coli / Q000187:drug effects* / Q000254:growth & development / Q000302:isolation & purification; D005672:Fusidic Acid / Q000494:pharmacology; D057166:High-Throughput Screening Assays / Q000295:instrumentation*; D056656:Lab-On-A-Chip Devices; D008826:Microbial Sensitivity Tests / Q000295:instrumentation*; D050296:Microbial Viability / Q000187:drug effects; D009154:Mutation; D010316:Particle Size; D000075082:Proof of Concept Study; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D059010:Single-Cell Analysis / Q000295:instrumentation; D000076227:Stereolithography
chemical_list: D000900:Anti-Bacterial Agents; D004655:Emulsions; D011500:Protein Synthesis Inhibitors; D005672:Fusidic Acid