AEM Accepts, published online ahead of print on 6 November 2009

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Appl. Environ. Microbiol. doi:10.1128/AEM.01125-09
Copyright (c) 2009, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Monitoring the dissemination of the broad host range plasmid pB10 in sediment microcosms by quantitative PCR

Sébastien Bonot and Christophe Merlin^*

Centre International de Recherche sur l'Eau et l'Environnement (CIRSEE - Suez Environnement), 38 rue du président Wilson, 78230 Le Pecq, France; LCPME, Laboratoire de Chimie Physique et Microbiologie pour l'Environnement, Nancy-Université, CNRS, 405 rue de Vandoeuvre, 54600 Villers-lès-Nancy, France

^* To whom correspondence should be addressed. Email: christophe.merlin{at}pharma.uhp-nancy.fr.

Studying the transfer of specific of mobile genetic elements in complex environmental matrices remains difficult because suitable molecular tools are not yet available to back up classical culture-dependent approaches. In this report we show that quantitative PCR could be used to monitor the dissemination of the broad host range plasmid pB10 in sediment microcosms. This approach lies on the differential measurement of the host and plasmid DNAs used to inoculate the microcosms, using a particular design of quantitative PCR primers/probes where we took advantage of the mosaic aspect of the bacterial genomes to achieve a highly specific quantitative PCR detection system.