4.7 Article

Facile electrochemical detection of Escherichia coli using redox cycling of the product generated by the intracellular β-D-galactosidase

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

  1. National Research Foundation of Korea [2010-0020780, 2012R1A2A2A06045327, 2012-M3C1A1-048860]
  2. National Research Foundation of Korea [2012R1A2A2A06045327, 2010-0020780] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)

向作者/读者索取更多资源

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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