Sensitive and Selective Detection of Enterococcus faecalis Using a New Turn-on Fluorogenic β-glucosidase Substrate Combined with a Modified Selective Broth
A new, simple-to-synthesize and sensitive turn-on fluorogenic substrate ( CFMU-Glu ) for β-glucosidase activity was developed. This probe was based on a 7-hydroxycoumarin derivative ( CFMU ) that could emit green fluorescence and had the low p K a value of 5.61 ± 0.01. CFMU-Glu could be used for sensitive monitoring of the almond βGLU and Enterococcus faecalis ( E. faecalis ) at the optimal pHs of 6.50 and 7.00, respectively. Moreover, a new sensitive and selective fluorogenic broth (PBF-B) for E. faecalis , utilizing CFMU-Glu and polymyxin B, was also developed. Polymyxin B was discovered to can significantly improve the detection selectivity and signal intensity. The proposed 4-four method using PBF-B and a microcentrifuge tube could provide fluorogenic detection limits of 5.01 × 10 4 and 1.0 × 10 5 CFU mL −1 by fluorescence microplate reader and naked eye, respectively; it could also provide a turn-on chromogenic detection limit of 1.0 × 10 6 CFU mL −1 by naked eye. The proposed method could detect 8 CFU mL −1 of E. faecalis in drinking water, Liangcha (herbal tea) and milk samples within 10 h, without pre-enrichment.
危险品化学品经营许可证（带存储）