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 pKa 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 × 104 and 1.0 × 105 CFU mL−1 by fluorescence microplate reader and naked eye, respectively; it could also provide a turn‐on chromogenic detection limit of 1.0 × 106 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. [ABSTRACT FROM AUTHOR]