Crystal Violet and XTT Assays on Staphylococcus aureus Biofilm Quantification
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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.
KeywordsExtracellular Polymeric Substance Crystal Violet Latex Agglutination Test Crystal Violet Assay Food Poisoning Case
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).
- 3.Christensen GD, Simpson W, Younger J, Baddour L, Barrett F, Melton D, Beachey E (1985) Adherence of coagulase-negative staphylococci to plastic tissue culture plates: a quantitative model for the adherence of staphylococci to medical devices. J Clin Microbiol 22(6):996–1006PubMedPubMedCentralGoogle Scholar
- 12.Gill SR, Fouts DE, Archer GL, Mongodin EF, DeBoy RT, Ravel J, Paulsen IT, Kolonay JF, Brinkac L, Beanan M (2005) Insights on evolution of virulence and resistance from the complete genome analysis of an early methicillin-resistant Staphylococcus aureus strain and a biofilm-producing methicillin-resistant Staphylococcus epidermidis strain. J Bacteriol 187(7):2426–2438CrossRefPubMedPubMedCentralGoogle Scholar
- 23.Silva WJd, Seneviratne J, Parahitiyawa N, Rosa EAR, Samaranayake LP, Cury AADB (2008) Improvement of XTT assay performance for studies involving Candida albicans biofilms. Braz Den J 19(4):364–369Google Scholar