Laser scanning detection of FISH-labelled Escherichia coli from water samples.
The development of rapid and accurate methods for the detection and quantification of bacteria without cultivation is of increasing importance for water monitoring. The aim of this study was to develop a solid phase cytometry detection method for DVC-FISH labelled Escherichia coli cells. In order to allow Stomatic detection with ChemScan RDI, the fluorescein-tyramide was combined with an oligonucleotide probe directly labelled with horseradish peroxidase to increase the fluorescence intensity. The method developed was tested for the enumeration of pure cultures, for GAC-filtered and drinking water samples. The method, which appeared to be equivalent to the culture method, was less sensitive than the DVC-FISH method followed by microscopic analysis. Research is underway to further optimise the labelling conditions.
Lepeuple
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Anjou Recherche - Vivendi Water, 1, Place de Turenne, 94417 Saint Maurice, France. anne-sophie.lepeuple@generale-des-eaux.net
Delabre
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Gilouppe
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Intertaglia
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de Roubin
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MR
D001331:Automation; D004784:Environmental Monitoring / Q000379:methods; D004926:Escherichia coli / Q000235:genetics* / Q000302:isolation & purification; D006735:Horseradish Peroxidase / Q000494:pharmacology; D017404:In Situ Hybridization, Fluorescence; D007834:Lasers; D015345:Oligonucleotide Probes; D014871:Water Microbiology; D014881:Water Supply
D015345:Oligonucleotide Probes; D006735:Horseradish Peroxidase