It is essential to have efficient, simple, quick and transportable tools for reliably identifying bacteria that are multiresistant to antibiotics, more specifically extended spectrum ?-lactamase (ESBL)-producing Enterobacteriaceae, which are the most widespread among Enterobacteriaceae. The present invention meets this requirement through its ease of use and its speed. The invention is based on detecting the enzyme activity of ?-lactam hydrolysis using an antibody capable of discriminating between the intact form of the ?-lactam ring of a ?-lactam and its hydrolysis product. This antibody can be used in kits and methods enabling for rapidly detecting (in less than one hour), without using expensive equipment (a small strip visible to the naked eye), the presence of bacteria producing penicillin-type, plasmid-mediated or hyper-produced AmpC enzymes, of ESBL or carbapenemase from colonies or in a sample.

Direct detection of mutagenesis in prokaryotes by reversion of an inactivating mutation (reversion mutation assay), producing a quantitative signal for in vivo mutagenesis, may greatly reduce the amount of test chemicals and labor involved in these assays. Further, transcriptional coupling of -lactamase reversion and GFP, translational fusion between -lactamase and GFP with stop codon in GFP, and a novel dual reporter to monitor continuous mutagenesis may be used in methods described herein.

The present invention provides a universal delivery platform of functional, heterologous compounds to specific cells using toxins modified to include a heterologous compound. In one embodiment, the toxin is an AB5 toxin. In one embodiment, the AB5 toxin is a heat-labile enterotoxin from E. coli (LT), including LTI, LTII, LTIIa, LTIIb, LTIIc and other recombinant forms of LT. Methods of use are also provided.

The present invention provides, in part, therapeutic beta-lactamases that can, inter alia, mitigate antibiotic resistance.

The present invention relates to the field of microbiology, in particular to the field of bacterial antibiotic resistance. more particularly to the field of resistance to beta-lactam inhibitors. The invention provides non-natural compounds which induce the aggregation of beta-lactamases, particularly beta-lactamases of class A. In addition, the invention provides combination therapies and pharmaceutical compositions between the non-natural molecules and beta-lactam antibiotics.

The invention relates to, in part, improved methods for the production of beta-lactamase using Escherichia coli (E. coli) cells. High yield production of beta-lactamase is achieved using methods of the invention.

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.

The present invention relates to pharmaceuticals and modified beta-lactamases. Specifically, the invention relates to novel recombinant beta-lactamases and pharmaceutical compositions comprising the beta-lactamases.

Also, the present invention relates to methods for modifying a beta-lactamase, producing the beta-lactamase and treating or preventing beta-lactam antibiotic induced adverse effects. Furthermore, the present invention relates to the beta-lactamase for use as a medicament and to the use of the beta-lactamase in the manufacture of a medicament for treating or preventing beta-lactam antibiotics induced adverse effects.

Still further, the invention relates to a polynucleotide and a host cell comprising the polynucleotide.

The disclosure provides constructs comprising a first fusion protein, a second fusion protein, and a linker, wherein the first fusion protein and the second fusion protein each include an affinity reagent and a reactive enzyme, and the linker includes a first and second functional groups specific for irreversibly inhibiting the first and second fusion protein reactive enzymes. The disclosure further provides a method including (a) contacting a first fusion protein including an affinity reagent and a reactive enzyme with a linker including a functional group specific for irreversibly inhibiting the first fusion protein reactive enzyme thereby coupling the first fusion protein and the linker, and (b) contacting a second fusion protein including an affinity reagent and a reactive enzyme with the linker, the linker including a functional group specific for irreversibly inhibiting the second fusion protein reactive enzyme thereby coupling the second fusion protein and the linker.