The detection of Staphylococcus aureus （S. aureus） is very important as it is responsible for bacterial infectious diseases and food poisoning. In this paper we explored the application of fluorescently labelled van...The detection of Staphylococcus aureus （S. aureus） is very important as it is responsible for bacterial infectious diseases and food poisoning. In this paper we explored the application of fluorescently labelled vancomycin to specifically bind and detect S. aureus. In view of the specificity of vancomycin towards bacterial cell surfaces, we utilized Cy5 to label vancomycin （CyS-Van） for the identification of S. aureus. Our experiments were designed to examine in greater detail the specificity of the reaction between CyS- Van and S. aureus. Detection parameters such as the derivatization conditions, concentrations of buffer, pH value, response performance of CyS-Van to bacterial surface, injection time and reversed-polarity time have been investigated and optimized. To develop a simple and quick assay for the detection of S. aureus at low concentrations, we propose to use the Cy5-Van for labeling the 5. aureus coupled with an on-line multiple-concentration in microcbip electrophoresis. Under the optimized conditions, the detection orS. aureus was achieved within 150 s with limit of detection （S/N = 3） of 981 CFU/mL, and 350- fold enhancement was obtained for S. aureus as compared to using the no concentration step. It is self- evident that this approach has great potential in the future for the analysis of S. aureus.展开更多
基金supported by the National Key R&D Program of China[grant number 2018YFC0809400]the National Natural Science Foundation of China[grant number 41975057].
基金supported by the National Natural Science Foundation of China(No.21575042)
文摘The detection of Staphylococcus aureus （S. aureus） is very important as it is responsible for bacterial infectious diseases and food poisoning. In this paper we explored the application of fluorescently labelled vancomycin to specifically bind and detect S. aureus. In view of the specificity of vancomycin towards bacterial cell surfaces, we utilized Cy5 to label vancomycin （CyS-Van） for the identification of S. aureus. Our experiments were designed to examine in greater detail the specificity of the reaction between CyS- Van and S. aureus. Detection parameters such as the derivatization conditions, concentrations of buffer, pH value, response performance of CyS-Van to bacterial surface, injection time and reversed-polarity time have been investigated and optimized. To develop a simple and quick assay for the detection of S. aureus at low concentrations, we propose to use the Cy5-Van for labeling the 5. aureus coupled with an on-line multiple-concentration in microcbip electrophoresis. Under the optimized conditions, the detection orS. aureus was achieved within 150 s with limit of detection （S/N = 3） of 981 CFU/mL, and 350- fold enhancement was obtained for S. aureus as compared to using the no concentration step. It is self- evident that this approach has great potential in the future for the analysis of S. aureus.
