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Published on: October 9, 2016
Optimization of processing conditions for the quantification of enterococci biofilms using microtitre-plates
C I Extremina1, L Costa, A I Aguiar
1REQUIMTE, Laboratório de Microbiologia, Faculdade de Farmácia, Universidade do Porto, Porto, Portugal. extremina@gmail.com
Journal of Microbiological Methods
|November 18, 2010
Summary
This study optimized a protocol for evaluating enterococci biofilm formation, enhancing accuracy and simulating real-world conditions for better biofilm screening. The new method allows for reproducible quantification using various assays and parameters.
Area of Science:
- Microbiology
- Biotechnology
Background:
- Enterococci biofilm formation is crucial for understanding infections.
- Existing protocols for biofilm assessment vary in accuracy and reproducibility.
- Standardized methods are needed for reliable enterococci biofilm evaluation.
Purpose of the Study:
- To compare and optimize protocols for assessing enterococci biofilm formation.
- To develop a dynamic model that better simulates in vivo conditions.
- To evaluate the reproducibility of different biofilm quantification methods.
Main Methods:
- Comparison of four literature-based protocols using microtitre-plate assays.
- Optimization of biofilm formation steps for stringent performance.
- Application of colorimetric biomass (crystal violet), resazurin, and CFU assays.
- Utilized biofilm formation index (BFi), cut-off values, and Z' factor for analysis.
Main Results:
- An optimized dynamic model protocol was developed for enhanced discrimination of enterococci biofilm formation.
- The optimized protocol demonstrated adequate reproducibility with multiple quantification approaches.
- Key parameters like BFi and Z' factor were identified for improved accuracy and quality control.
Conclusions:
- The optimized protocol offers a more accurate and reproducible method for enterococci biofilm assessment.
- The dynamic model better mimics in vivo conditions, aiding clinical relevance.
- Standardized parameters enhance inter-laboratory comparisons and assay quality evaluation.
