期刊
SENSORS AND ACTUATORS B-CHEMICAL
卷 209, 期 -, 页码 951-956出版社
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2014.12.073
关键词
E. coli; beta-D-Galactosidase; Redox cycling; Electrochemical detection
资金
- National Research Foundation of Korea [2010-0020780, 2012R1A2A2A06045327, 2012-M3C1A1-048860]
- National Research Foundation of Korea [2012R1A2A2A06045327, 2010-0020780] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
Various detection methods for pathogenic bacteria have been developed, most of which are based on cell culture, DNA amplification, and immunoassays. Although the methods allow highly sensitive and/or selective detection, they require long detection periods and complex detection procedures. This paper reports a facile electrochemical detection method for Escherichia coli (E. coli) without the use of DNA amplification or immunoassays. The detection method harnesses the intracellular beta-D-galactosidase (Gal) activity of E. coli along with the signal amplification based on redox cycling. The Gal expression level is increased by treatment with a Gal expression-inducer (isopropyl-beta-D-thiogalactopyranoside; IPTG); the enzymatic reaction of Gal is facilitated by the permeabilization treatment involving the use of chloroform and sodium dodecyl sulfate; the electrochemical signal is amplified by the electrochemical-chemical-chemical (ECC) and chemical-chemical redox cycling involving the Gal product, Ru(NH3)(6)(3+) and tris(2-carboxyethyl)phosphine. The detection limit obtained in the presence of the ECC redox cycling, with a total detection period of only 4.5 h, is ca. 1 colony forming unit (CFU)/mL, indicating that ECC redox cycling allows for a sensitive detection. (C) 2014 Elsevier B.V. All rights reserved.
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