ABSTRACT
Bacterial resistance caused by ß-lactamases has been a major threat to public health around the world, seriously weakening the efficacy of ß-lactam antibiotics, the most widely used therapeutic agents against infectious diseases. To detect the bacterial resistance to ß-lactam antibiotics, particularly specific type of ß-lactam antibiotics, in a rapid manner, we report herein a relay-response chemiluminescence assay. This assay mainly consists of two reagents: a ß-lactam-caged thiophenol and a thiophenol-sensitive chemiluminescence reporter, both of which are synthetically feasible. The selective hydrolysis of ß-lactam by ß-lactamase leads to the releasing of free thiophenol, which then triggers the emission of a chemiluminescence signal in a relay manner. Three thiophenol-caged ß-lactams, structural analogues of cephalothin, cefotaxime, and meropenem, respectively, have been synthesized. And the application of this assay with these analogues of ß-lactam antibiotics allows fast detection of ß-lactamase-expressing resistant bacteria and, more impressively, provides detailed information on the resistant scope of bacteria.
Subject(s)
Anti-Bacterial Agents , Luminescent Measurements , beta-Lactam Resistance , beta-Lactamases , beta-Lactams , beta-Lactams/pharmacology , Anti-Bacterial Agents/pharmacology , Luminescent Measurements/methods , beta-Lactamases/metabolism , Microbial Sensitivity Tests , Bacteria/drug effects , beta Lactam AntibioticsABSTRACT
A fluorogenic probe for the specific detection of IMP-1 ß-lactamase activity has been developed. This imaging reagent features a unique trans-acetylamino cephalosporin as an enzymatic recognition moiety, exhibiting excellent selectivity to IMP-1 ß-lactamase over other ß-lactamases, including serine- and metallo-ß-lactamases. The selective activation of the probe by IMP-1 ß-lactamase leads to over 30-fold enhancement in the fluorescence intensity, which allows enzyme activity to be reported with high sensitivity.
