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Applied and Environmental Microbiology, July 2008, p. 4054-4058, Vol. 74, No. 13
0099-2240/08/$08.00+0     doi:10.1128/AEM.02806-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Surface Plasmon Resonance Assay for Real-Time Monitoring of Somatic Coliphages in Wastewaters{triangledown}

Cristina García-Aljaro,1* Xavier Muñoz-Berbel,1 A. Toby A. Jenkins,2 Anicet R. Blanch,3 and Francesc Xavier Muñoz1

Centre Nacional de Microelectrònica de Barcelona (CNM-IMB), CSIC, Esfera UAB, Campus UAB, 08193 Bellaterra, Spain,1 Department of Chemistry, University of Bath, Bath BA2 7AY, United Kingdom,2 Departament de Microbiologia, Facultat de Biología, Universitat de Barcelona, 08028 Barcelona, Spain3

Received 12 December 2007/ Accepted 5 May 2008

The surface plasmon resonance (SPR) technique is a well-established method for the measurement of molecules binding to surfaces and the quantification of binding constants between surface-immobilized proteins and proteins in solution. In this paper we describe an extension of the methodology to study bacteriophage-bacterium interactions. A two-channel microfluidic SPR sensor device was used to detect the presence of somatic coliphages, a group of bacteriophages that have been proposed as fecal pollution indicators in water, using their host, Escherichia coli WG5, as a target for their selective detection. The bacterium, E. coli WG5, was immobilized on gold sensor chips using avidin-biotin and bacteriophages extracted from wastewater added. The initial binding of the bacteriophage was observed at high concentrations, and a separate, time-delayed cell lysis event also was observed, which was sensitive to bacteriophage at low concentrations. As few as 1 PFU/ml of bacteriophage injected into the chamber could be detected after a phage incubation period of 120 min, which equates to an approximate limit of detection of around 102 PFU/ml. The bacteriophage-bacterium interaction appeared to cause a structural change in the surface-bound bacteria, possibly due to collapse of the cell, which was observed as an increase in mass density on the sensor chip. These results suggest that this methodology could be employed for future biosensor technologies and for quantification of the bacteriophage concentration.

* Corresponding author. Mailing address: Centre Nacional de Microelectrònica de Barcelona (CNM-IMB), CSIC, Esfera UAB, Campus UAB, 08193 Bellaterra, Spain. Phone: 34 935947700, ext. 2119. Fax: 34 935801496. E-mail: cristina.garcia{at}cnm.es

{triangledown} Published ahead of print on 9 May 2008.

Applied and Environmental Microbiology, July 2008, p. 4054-4058, Vol. 74, No. 13
0099-2240/08/$08.00+0     doi:10.1128/AEM.02806-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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