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Automated fluorescent in situ hybridization for the specific detection and quantification of oral streptococci in dental plaque.
- Source :
-
Journal of microbiological methods [J Microbiol Methods] 2001 Feb 01; Vol. 44 (1), pp. 39-47. - Publication Year :
- 2001
-
Abstract
- Our aim was to develop a rapid fluorescent in situ hybridization (FISH) assay for the identification of different oral groups of streptococci in dental plaque and to combine it with digital image analysis for the automated enumeration of target cells. Cy3-labeled oligonucleotide probes specific for 16S rRNA gene sequences of the anginosus, mitis, mutans, and salivarius groups of streptococci were hybridized under stringent conditions with bacterial cultures or supragingival plaque samples that had been permeabilized with lysozyme. Probe specificity was determined with strains from 30 different species, mainly of oral origin. Results showed that probes ANG541, MIT447, SSP001, and SAL090 with specificity for the anginosus, mitis, mutans, and salivarius groups, respectively, the pan-reactive streptococcal probe STR405, the S. mutans specific probe MUT590, and the S. sobrinus specific probe SOB174 were well-suited for the identification of cultured streptococci. Probes STR405, MIT447 and SSP001 were then successfully applied to enumerate automatically bacteria of the recognized taxa in 144 supragingival plaque samples. On the average, total streptococci accounted for 8.2%, streptococci of the mitis and mutans groups for 3.9 and 1.7%, respectively, of the plaques. The combined application of FISH and automated image analysis provides an objective time-saving alternative to culture or PCR for the enumeration of selected oral streptococci in dental plaque.
- Subjects :
- Colony Count, Microbial
DNA, Bacterial analysis
Humans
Image Processing, Computer-Assisted
In Situ Hybridization, Fluorescence
Oligonucleotide Probes chemical synthesis
Streptococcus cytology
Streptococcus genetics
Dental Plaque microbiology
Mouth microbiology
Streptococcus isolation & purification
Subjects
Details
- Language :
- English
- ISSN :
- 0167-7012
- Volume :
- 44
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Journal of microbiological methods
- Publication Type :
- Academic Journal
- Accession number :
- 11166098
- Full Text :
- https://doi.org/10.1016/s0167-7012(00)00226-8