Presented herein are methods and compositions for the detection of specific beta-lactamases, including class A serine carbapenemases, metallo-beta-lactamases, AmpC beta-lactamases, and extended-spectrum beta-lactamases (ESBLs). The methods presented herein include methods that permit the detection of the presence of specific beta-lactamases in bacterial samples within as few as 2 to 10 minutes.

Antimicrobial resistance (AMR), the ability of a bacterial species to resist the action of an antimicrobial drug, has been on the rise due to the widespread use of antimicrobial agents, and one of the many ways AMR can spread is through contaminated water sources. To monitor these water sources, we have developed an inexpensive, fast assay using a paper-based analytical device (PAD) that can test for the presence of -lactamase-mediated resistance as one major form of AMR that has reliably detected resistance in sewage water.

The present invention discloses a probe useful for the selective detection of metallo-beta-lactamases, in particular carbapenemases, thereby distinguishing those species of bacteria that are carbapenem-resistant from bacterial strains that are sensitive. Cephalospori based probes that have the 6,7 R,R configuration are susceptible to cleavage by beta-lactamases but cannot distinguish between cleavage by metallo-beta-lactamases and other beta-lactamases. By modifying a side group of the cephalosporin, selectivity can be introduced allowing the probes to distinguish between various types of metallo-beta-lactamases.

Based on the discovery that MBLAC1 is a specific, high-affinity target for Ceftriaxone (Cef), MBLAC1 may be used for identifying treatments for addiction to substances of abuse. Methods for identifying therapeutic agents for treatment of addiction to a substance of abuse include using an assay to determine if a test agent is capable of binding to MBLAC1 or disrupting binding between MBLAC1 protein and Cef, and identifying such a test agent as a candidate therapeutic agent for treatment of addiction to a substance of abuse. MBLAC knock-out (KO) animals, methods of use thereof, and kits are used for identifying a therapeutic agent that reduces the actions of at least one substance of abuse. Methods also include using cellular extracts from tissue or cultured cells taken from wild-type (WT) MBLAC1 and MBLAC1 KO animals for screening for novel, Cef-like molecules in vitro, and using cells from a MBLAC1 KO animal to test for Cef-like actions of a test molecule.

The present disclosure features portable wide field fluorimeter systems, e.g., in the form of low-cost mobile platforms, and methods to perform fluorometric assays to detect a change in fluorescence intensity in liquid samples, e.g., caused by the presence of a target analyte, e.g., a protein, e.g., an enzyme (e.g., -lactamase) expressed by a target pathogen in a liquid sample in a point-of-care setting. In some implementations, a portable system for detecting a change in fluorescence intensity in a liquid sample includes a microfluidic device, an optical assembly including an emission filter and one or more lenses, and an analyzer device that collects and processes a fluorescent signal for the detection of a target analyte produced by the target pathogen present in the liquid sample.

A method for assessing drug-resistant Klebsiella pneumoniae includes the following steps. A test sample is provided, wherein the test sample includes a Klebsiella pneumoniae. A spectrum analysis step is performed, wherein the test sample is detected by a mass spectrometry method so as to obtain a target mass spectrum data. An assessing step for drug-resistant Klebsiella pneumoniae is performed, wherein the target mass spectrum data is analyzed so as to assess whether the Klebsiella pneumoniae is resistant to a carbapenem antibiotic or a colistin or not. When the Klebsiella pneumoniae is resistant to the carbapenem antibiotic, the target mass spectrum data includes a first anti-carbapenem feature mark, and when the Klebsiella pneumoniae is resistant to the colistin, the target mass spectrum data includes a first anti-colistin feature mark.

A method of calibrating a device for measuring the concentration of creatinine using one or more calibration solutions, the method comprising: receiving concentrations at an initial time of creatine, Cr, and/or creatinine, Crn, of the one or more calibration solutions; receiving outputs of the measuring device at the end time; calculating the concentration of Cr and/or Crn in the calibration solutions at an end time using a temperature model, wherein the temperature model indicates changes in temperature of the calibration solutions from the initial time to the end time; and determining a relationship between the outputs of the measuring device and the calculated concentrations of Cr and/or Crn.

In an analysis method of the present invention, a compound represented by the following formula (I) as a matrix is mixed into an analysis target sample and the mixture is subjected to matrix-assisted laser desorption ionization mass spectrometry. In the formula (I), R is an alkyl group having 3-11 carbon atoms. The analysis target sample is a substance for which whether or not -lactamase is contained is to be determined. Analysis targets include, for example, bacteria and an, extract from bacteria.

##STR00001##

A compound which is a thienolate of formula (I) or a pharmaceutically acceptable salt thereof: (I) wherein R.sup.1, R.sup.3, Ring A1, n and Ring A2 are as defined herein, are found to be useful in inhibiting metallo-beta-lactamase and therefore in potentiating the activity of beta lactamase antibiotics. The compound can be used alone or in combination with a rhodanine of formula (II) or a pharmaceutically acceptable salt thereof: (II) wherein R.sup.3, Ring A1, n, Ring A2, L and Ring B are as defined herein. Treatment or prevention of bacterial infection in combination with beta-lactam antibiotic agents is also provided. ##STR00001##

Phenotypic antimicrobial susceptibility testing (AST), the gold-standard diagnostic that indicates whether an antimicrobial will be clinically effective, often suffer the slowest times-to-result for the most resistant pathogens. Here we introduce novel assays to be performed in parallel with standard AST assays that enable rapid, same-shift reporting of AST results for a plurality of pathogens. The assays developed here are further capable of detecting resistance to carbapenems, the most powerful class of beta-lactams commonly used as last-resort antimicrobials.