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Broad‐spectrum protein biosensors for class‐specific detection of antibiotics
Author(s) -
Weber Cornelia C.,
Link Nils,
Fux Cornelia,
Zisch Andreas H.,
Weber Wilfried,
Fussenegger Martin
Publication year - 2004
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.20224
Subject(s) - antibiotics , biosensor , tetracycline , tetracycline antibiotics , bacteria , computational biology , biology , broad spectrum , dna , chemistry , microbiology and biotechnology , biochemistry , combinatorial chemistry , genetics
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. © 2004 Wiley Periodicals, Inc.