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Appl. Environ. Microbiol., Aug 1997, 3274-3278, Vol 63, No. 8
Copyright © 1997, American Society for Microbiology

A New Sensitive, Whole-Cell Hybridization Technique for Detection of Bacteria Involving a Biotinylated Oligonucleotide Probe Targeting rRNA and Tyramide Signal Amplification

P Lebaron, P Catala, C Fajon, F Joux, J Baudart and L Bernard
Observatoire Oceanologique, Universite Pierre et Marie Curie, Institut des Sciences de l'Univers, CNRS URA2071, F-66651 Banyuls-sur-Mer, France

A tyramide signal amplification system with biotinylated oligonucleotide probes and streptavidin-horseradish peroxidase was used to increase the sensitivity of fluorescent in situ hybridization techniques. When applied to both gram-negative and -positive bacteria immobilized on glass slides, a 7- to 12-fold amplification of the fluorescence signal was observed relative to that of cells hybridized with fluorescently monolabeled probes. A large proportion (62 to 78%) of bacteria could be detected under starvation conditions and in natural samples from the marine environment. This amplification procedure allows new investigations in marine oligotrophic ecosystems and water quality control.


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