Single-cell identification by improved fluorescence in situ hybridisation techniques
Abstract number: S487
Amann R.I., Fuchs B.M.
The ribosomal RNA (rRNA) approach to microbial evolution and ecology has become an integral part of microbiology. Rapidly growing databases exist that encompass besides the 16S rRNA sequences of almost all validly described Bacteria and Archaea also numerous 16S rRNA sequences of so far uncultivated microbes, directly retrieved from the environment by PCR or metagenomics. Based on the patchy evolutionary conservation of rRNA genes oligonucleotide probes can be designed in a directed way with specificities ranging from species up to large evolutionary entities like phyla or even domains.
When such probes are labeled with fluorescent dyes or the enzyme horseradish peroxidase they can be used to identify single microbial cells by fluorescence in situ hybridisation (FISH) directly in complex environmental samples. An update on recent applications and methodological improvements will be given which includes the identification of small bacterial cells by catalyzed reporter deposition (CARD)-FISH. With optimised methods and proper controls FISH yields exact cell numbers and spatial distributions for defined bacterial populations also in highly complex mixed microbial communities.
|Session name:||19th European Congress of Clinical Microbiology and Infectious Diseases|
|Location:||Helsinki, Finland, 16 - 19 May 2009|
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