title: Broad-spectrum protein biosensors for class-specific detection of antibiotics.
abstract: The dramatically increasing prevalence of multi-drug-resistant human pathogenic bacteria and related mortality requires two key actions: (i) decisive initiatives for the detection of novel antibiotics and (ii) a global ban for use of antibiotics as growth promotants in stock farming. Both key actions entail technology for precise, high-sensitive detection of antibiotic substances either to detect and validate novel anti-infective structures or to enforce the non-use of clinically relevant antibiotics. We have engineered prokaryotic antibiotic response regulators into a molecular biosensor configuration able to detect tetracycline, streptogramin, and macrolide antibiotics in spiked liquids including milk and serum at ng/mL concentrations and up to 2 orders of magnitude below current Swiss and EC threshold values. This broad-spectrum, class-specific, biosensor-based assay has been optimized for use in a storable ready-to-use and high-throughput-compatible ELISA-type format. At the center of the assay is an antibiotic sensor protein whose interaction with specific DNA fragments is responsive to a particular class of antibiotics. Binding of biosensor protein to the cognate DNA chemically linked to a solid surface is converted into an immuno-based colorimetric readout correlating with specific antibiotics concentrations.
authors.affiliation: Institute for Biomedical Engineering, Swiss Federal Institute of Technology, ETH Zurich, CH-8044 Zurich, Switzerland.
authors.forename: Cornelia C
authors.initials: CC
authors.lastname: Weber
authors.forename: Nils
authors.initials: N
authors.lastname: Link
authors.forename: Cornelia
authors.initials: C
authors.lastname: Fux
authors.forename: Andreas H
authors.initials: AH
authors.lastname: Zisch
authors.forename: Wilfried
authors.initials: W
authors.lastname: Weber
authors.forename: Martin
authors.initials: M
authors.lastname: Fussenegger
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis* / Q000494:pharmacology; D001419:Bacteria / Q000187:drug effects / Q000472:pathogenicity; D001426:Bacterial Proteins; D015374:Biosensing Techniques; D015153:Blotting, Western; D016466:CHO Cells; D003124:Colorimetry; D006224:Cricetinae; D003412:Cricetulus; D004305:Dose-Response Relationship, Drug; D024901:Drug Resistance, Multiple, Bacterial; D004359:Drug Therapy, Combination; D004797:Enzyme-Linked Immunosorbent Assay; D017930:Genes, Reporter; D006801:Humans; D010455:Peptides; D011506:Proteins / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D012680:Sensitivity and Specificity
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D010455:Peptides; D011506:Proteins