Fluorogenic assay for rapid screening of bacterial beta-lactamase activity
AssigneeThe Board of Trustees of the Leland Stanford Junior University
InventorsJianghong RAO, Jinghang XIE, Ran MU
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Abstract
A dual-caged fluorogenic resorufin probe (CDA) has been developed that is stable under physiological condition with low background but becomes highly fluorescent upon β-lactamase/esterase activation and further oxidation. The probes of the disclosure are advantageous for initial screening of broad-spectrum β-lactam antibiotics resistance and carbapenem resistant pathogens at diagnosis. After a two-step filtration, the assay of the disclosure can report 103 c.f.u./mL cephalosporin- and carbapenem-resistant bacteria in urine within 2 hours at room temperature.
Claims18
A dual-caged cleavable probe comprising a b-lactamase- or carbapenemase-cleavable moiety conjugated to a self-immolative linker, wherein the self-immolative linker is conjugated to a fluorescently detectable label.
A method of detecting the presence of a bacterial strain that has resistance to at least one of a beta-lactam antibiotic or a carbapenem antibiotic, wherein said method comprises contacting a population of bacteria suspected of being resistant to at least one of a beta-lactam antibiotic or a carbapenem antibiotic with a dual-caged N-cephalosporin-caged 3,7-diesterphenoxazine beta-lactamase-cleavable probe or a dual-caged carbapenem-caged 3,7-diesterphenoxazine carbapenem-cleavable probe in the presence of an oxidizing agent, and measuring a fluorescent signal, wherein a detectable fluorescent signal indicates that the bacterial strain has at least one of a beta-lactamase or a carbapenemase.
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Dependencies are inferred from the claim text.