Amperometric detection of Enterobacteriaceae in river water by measuring β-galactosidase activity at interdigitated microelectrode arrays.

Olivier Laczka, Cristina García-Aljaro, Francisco Javier del Campo, Francesc Xavier Muñoz Pascual, Jordi Mas-Gordi, Eva Baldrich

Index: Anal. Chim. Acta 677 , 156-161, (2010)

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Abstract

Two simple methodologies are compared for the detection of faecal contamination in water using amperometry at gold interdigitated microelectrodes. They rely on the detection of β-galactosidase (β-gal) by redox cycling amperometry of the p-aminophenol (PAP) produced by the enzyme from the 4-aminophenyl β-d-galactopyranoside (PAPG) substrate. The use of phages as specific agents for the release of the bacteria-enclosed enzyme allowed the detection of 6×10(5) CFU mL(-1)Escherichia coli in 2 h without any pre-enrichment or preconcentration steps. Better limits of detection were achieved for the second strategy in the absence of phages. In this case, bacteria were enriched in the presence of both β-d-1-thiogalactopyranoside (IPTG) and substrate but in the absence of phages. Under such experimental conditions, 5×10(4) CFU mL(-1) E. coli could be detected after 2 h of incubation, while 7 h of incubation were enough to detect down to 10 CFU mL(-1) in river water samples. This represents a straightforward one-step method for the detection of faecal contamination that can be conducted in a single working day with minimal sample manipulation by the user.Copyright © 2010 Elsevier B.V. All rights reserved.