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Evaluation of different microtiter plate-based methods for the quantitative assessment of Staphylococcus aureus biofilms

    Walid F Elkhatib

    Department of Microbiology & Immunology, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt

    Department of Pediatrics, Eastern Virginia Medical School, Norfolk, VA, USA

    Department of Pharmacy Practice, School of Pharmacy, Hampton University, Hampton, VA, USA

    ,
    Ahmed S Khairalla

    Department of Microbiology & Immunology, Faculty of Pharmacy, Beni-Suef University, Beni-Suef, Egypt

    &
    Hossam M Ashour

    Department of Pharmacy Practice, Eugene Applebaum College of Pharmacy & Health Sciences, Wayne State University, Detroit, MI, USA

    Department of Microbiology & Immunology, Faculty of Pharmacy, Cairo University, Cairo, Egypt

    Published Online:https://doi.org/10.2217/fmb.14.33

    ABSTRACT: 

    Aim: To quantitatively assess Staphylococcus aureus biofilms. Materials & methods: In addition to the qualitative Congo red agar (CRA) method, we used the bioluminescence (BLM), safranine (SAF), crystal violet (CRV) and resazurin (RES) high-throughput microtiter plate-based quantitative assays. Results: 60.47% (26/43) of S. aureus clinical isolates were weak biofilm producers. The CRA method detected positive-slime phenotypes (13.95%), but was unable to distinguish weak from negative producers. BLM assays demonstrated significant correlations with RES (highest), CRV and SAF (lowest). Lower coefficient of variation values indicate precision. BLM scored highest precision (coefficient of variation = 0.013) followed by RES, SAF and CRV. Conclusion: BLM and RES detect live biomass in S. aureus biofilms (for physiological studies). SAF and CRV detect live/dead bacteria plus biofilm matrix (for monitoring overall biofilm architecture, not only its cell viability). Reliable assays are essential for effective biofilm therapy.

    Papers of special note have been highlighted as: •• of considerable interest

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