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Détection de Vibrio cholerae dans un écosystème marin par hybridation moléculaire après culture sur un milieu sélectif

Abstract : A rapid and efficient molecular method based on culture selective medium and colony hybridization assay was developed to identify V. cholerae in water samples of Rance estuary. A 22 nucleotide sequence of the 16S-23S rDNA region was labeled with digoxigenin and evaluated for specificity and sensitivity by dot blot and colony hybridization with strains from the Collection de l'Institut Pasteur and environmental and clinical isolates of V. cholerae from the collection of the Centre National de Référence des Vibrions et du Choléra. The probe hybridized with all strains of V. cholerae. No isolates of species other than V. cholerae or other genera hybridized with the probe. This molecular technique allows specific detection of V. cholerae strains in mixed microbial populations from water samples, after enrichment procedure and growth on selective culture media, TCBS agar or mGNA, less selective but favoring the growth of this species. Moreover, this method increases the recovery of V. cholerae (10-fold higher) and can be used for its detection in natural samples after direct plating on mGNA, without prior enrichment. By this way, the laborious classic approach to control bacteria contamination in environmental marine water and seafood could be simplified.
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https://hal.ehesp.fr/hal-03315477
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Submitted on : Thursday, August 5, 2021 - 4:08:10 PM
Last modification on : Friday, August 6, 2021 - 3:34:53 AM

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Annick Robert-Pillot, Sandrine Baron, Jean Lesné, J.-M. Fournier, Marie-Laure Quilici. Détection de Vibrio cholerae dans un écosystème marin par hybridation moléculaire après culture sur un milieu sélectif. Hydroécologie Appliquée, EDP Sciences, 2006, 15, pp.97-105. ⟨10.1051/HYDRO:2006004⟩. ⟨hal-03315477⟩

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