ISSN:
1574-6968
Source:
Blackwell Publishing Journal Backfiles 1879-2005
Topics:
Biology
Notes:
The presence of conserved and variable regions in the 16S ribosomal RNA (rRNA) allows construction of oligonucleotide probes complementary to defined regions of this molecule. In this paper we apply such probes to identify members of Enterobacteriaceae, specifically closely related species of Proteus. Genus- and species-specific detection of Proteus sp. was done by dot-blotting, subspecies identification by Southern hybridization. Synthetic oligodeoxynucleotides, 24 to 29 bases in length, detected less than 103 bacteria in DNA/RNA dot-blots. Due to the favorable reaction kinetics, hybridization and washing times could be reduced to 15–20 min and 2–3 min, respectively. Sample preparation was critical, especially in the case of sputum specimens which were solubilized prior to RNA extraction. To prevent degradation of rRNA by the endogenous nuclease activity, the material was applied immediately to the filters. Otherwise, up to 1000-fold reduction in the hybridization signal was observed.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1111/j.1574-6968.1987.tb02121.x
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