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Crystal Violet and XTT Assays on Staphylococcus aureus Biofilm Quantification

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Abstract

Staphylococcus aureus (S. Aureus) is a common food-borne pathogenic microorganism. Biofilm formation remains the major obstruction for bacterial elimination. The study aims at providing a basis for determining S. aureus biofilm formation. 257 clinical samples of S. aureus isolates were identified by routine analysis and multiplex PCR detection and found to contain 227 MRSA, 16 MSSA, 11 MRCNS, and 3 MSCNS strains. Two assays for quantification of S. aureus biofilm formation, the crystal violet (CV) assay and the XTT (tetrazolium salt reduction) assay, were optimized, evaluated, and further compared. In CV assay, most isolates formed weak biofilm 74.3 %), while the rest formed moderate biofilm (23.3 %) or strong biofilm (2.3 %). However, most isolates in XTT assay showed weak metabolic activity (77.0 %), while the rest showed moderate metabolic activity (17.9 %) or high metabolic activity (5.1 %). In this study, we found a distinct strain-to-strain dissimilarity in terms of both biomass formation and metabolic activity, and it was concluded from this study that two assays were mutual complementation rather than being comparison.

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Acknowledgments

We gratefully acknowledge the National 973-Plan of China (2012CB720800), the National Natural Science Foundation of China (31201362 & 31101278), the International Science & Technology Cooperation Program (2013B051000014), the National Science and Technology Support Program (2012BAD37B01), the National Outstanding Doctoral Dissertation Funding (201459), the Guangdong Outstanding Doctoral Dissertation Funding (K3140030) andthe Open Project Program of State Key Laboratory of Food Science and Technology, Jiangnan University (Dr. Zhenbo Xu, 2015).

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Authors and Affiliations

  1. College of Food Science and Technology, South China University of Technology, Guangzhou, 510640, China

    Zhenbo Xu, Yanrui Liang, Shiqi Lin, Bing Li, Lin Li & Yang Deng

  2. Department of Biomedical Science, University of Maryland, Baltimore, MD, 21201, USA

    Zhenbo Xu

  3. Department of Laboratory Medicine, First Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510120, China

    Dingqiang Chen

  4. College of Light Industry and Food Science, South China University of Technology, 381 Wushan Road, Guangzhou, 510640, China

    Zhenbo Xu

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  1. Zhenbo Xu
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  2. Yanrui Liang
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  3. Shiqi Lin
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  5. Bing Li
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  6. Lin Li
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Correspondence to Zhenbo Xu.

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Xu, Z., Liang, Y., Lin, S. et al. Crystal Violet and XTT Assays on Staphylococcus aureus Biofilm Quantification. Curr Microbiol 73, 474–482 (2016). https://doi.org/10.1007/s00284-016-1081-1

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  • DOI: https://doi.org/10.1007/s00284-016-1081-1

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