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An Analytical Tool-box for Comprehensive Biochemical, Structural and Transcriptome Evaluation of Oral Biofilms Mediated by Mutans Streptococci
Published on: January 26, 2011
A novel spectrofluorometric microassay for Streptococcus mutans adherence to hydroxylapatite
1Education and Research Centre, St. Vincent's University Hospital, Elm Park, 4 Dublin, Ireland.
Journal of Microbiological Methods
|July 5, 2003
Summary
This study developed a novel, high-throughput fluorescence assay to measure Streptococcus mutans adherence to hydroxylapatite, offering a faster alternative for dental caries research.
Area of Science:
- Microbiology
- Biochemistry
- Dental Research
Background:
- Streptococcus mutans adherence to teeth initiates dental caries.
- Current methods for studying bacterial adherence are slow and may use radiolabels.
Purpose of the Study:
- Develop a convenient, high-throughput microtitre-plate assay for bacterial adherence.
- Utilize fluorescence-based detection for Streptococcus mutans adherence to hydroxylapatite.
Main Methods:
- Streptococcus mutans labelled with fluorescent indicator BCECF/AM.
- Optimized labelling conditions and adherence assay parameters (time, bacterial concentration).
- Measured fluorescence of bound bacteria to hydroxylapatite using a spectrofluorometer.
Main Results:
- Optimal labelling achieved with 50 microM BCECF/AM in DMSO for 120 min.
- Bacterial viability maintained post-labelling.
- Adherence reached saturation by 30 min with 1x10(7) cfu/ml labelled bacteria per mg hydroxylapatite.
- Assay demonstrated high reproducibility and ability to detect interference.
Conclusions:
- A convenient, fluorescence-based microtitre-plate assay for streptococcal adherence was developed.
- This assay is suitable for high-throughput screening of potential anti-adhesion agents.
- The method provides a valuable tool for dental caries research.
