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Trending unidentified bacteria using partial 16S rDNA sequencing



Trending unidentified bacteria using partial 16S rDNA sequencing



Abstracts of the General Meeting of the American Society for Microbiology 103: C-200



The ability to trend and track microbial contaminants is an important component of pharmaceutical manufacturing. This is traditionally accomplished by routine environmental monitoring of production areas, and identification of isolates using phenotypic methods. The resulting data (Genus and/or species assignments) are used to determine trends in the manufacturing environment, and locate the source of contamination. This approach has several limitations. Many environmental organisms are not identified accurately or reproducibly by conventional phenotypic methods, and organisms are often isolated that have not yet been described, resulting in no genus or species level identification. We determined the partial 16S rDNA sequence of 100 environmental contaminants that were previously unidentified in our laboratory using the MIDI Sherlock system, to determine whether accurate species level assignments could be made. 16S rDNA sequencing allowed us to identify 85% of the isolates to the genus level, and 40% of these isolates to the species level. Fifteen percent of the isolates were considered no match to any organism in the MicroSeq database. Further investigation revealed that 16S rDNA sequence data may be more appropriate for trending and tracking of organisms than taxonomic classification.

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