Abstract
The alarming increase of antimicrobial resistance urges rapid diagnosis and pathogen specific infection management. This work reports a rapid screening assay for pathogenic bacteria resistant to lactam antibiotics. We designed a fluorogenic N-cephalosporin caged 3,7-diesterphenoxazine probe CDA that requires sequential activations to become fluorescent resorufin. A series of studies with recombinant β-lactamases and clinically prevalent pathogens including Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae and Serratia marcescens demonstrated that CDA possessed superior sensitivity in reporting the activity of β-lactamases including cephalosporinases and carbapenemases. After a simple filtration, lactam-resistant bacteria in urine samples could be detected at 103 colony-forming units per milliliter within 2 hours.
Highlights
Jinghang Xie, ‡a Ran Mu,‡a Mingxi Fang,a Yunfeng Cheng,a Fiona Senchyna,b Angel Moreno,b Niaz Banaeibcd and Jianghong Rao *a
This work reports a rapid screening assay for pathogenic bacteria resistant to lactam antibiotics
A series of studies with recombinant blactamases and clinically prevalent pathogens including Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae and Serratia marcescens demonstrated that CDA possessed superior sensitivity in reporting the activity of b-lactamases including cephalosporinases and carbapenemases
Summary
Jinghang Xie, ‡a Ran Mu,‡a Mingxi Fang,a Yunfeng Cheng,a Fiona Senchyna,b Angel Moreno,b Niaz Banaeibcd and Jianghong Rao *a. We report a dual-caging design of a novel N-cephalosporin caged 3,7-diesterphenoxazine probe named CDA (Cephalosporin caged Diester Amplex red analogue) (Fig. 1a) and the development of a uorogenic assay for detecting blactamase expressing bacteria within hours of sample collection.
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