Kits for the detection of beta-lactamases
Presented herein are kits 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 kits presented herein include kits that permit the detection of the presence of specific beta-lactamases in bacterial samples within as few as 2 to 10 minutes.
1. A kit for detecting the presence of a beta-lactamase, comprising, in one or more containers:
(a) a first composition comprising a detectable beta-lactamase substrate and an AmpC inhibitor; and
(b) a second composition comprising a detectable beta-lactamase substrate, an AmpC inhibitor, and a serine beta-lactamase inhibitor in an amount sufficient to inhibit an extended-spectrum beta-lactamase (ESBL) and an original-spectrum beta-lactamase (OSBL) but not a class A serine carbapenemase.
2. A kit for detecting the presence of a beta-lactamase, comprising, in one or more containers:
(a) a first composition comprising a detectable beta-lactamase substrate and an AmpC inhibitor;
(b) a second composition comprising a detectable beta-lactamase substrate, an AmpC inhibitor, and a serine beta-lactamase inhibitor in an amount sufficient to inhibit an ESBL and an OSBL but not a class A serine carbapenemase; and
(c) a third composition comprising a detectable beta-lactamase substrate, an AmpC inhibitor, a serine beta-lactamase inhibitor in amount sufficient to inhibit an ESBL and an OSBL but not a class A serine carbapenemase, and a metal chelator.
3. The kit of claim 2 further comprising a fourth composition comprising a detectable beta-lactamase substrate and a serine beta-lactamase inhibitor in an amount sufficient to inhibit an ESBL and an OSBL but not a class A serine carbapenemase.
4. A kit for detecting the presence of a beta-lactamase, comprising, in one or more containers:
(a) a first composition comprising a detectable beta-lactamase substrate and an AmpC inhibitor;
(b) a second composition comprising a detectable beta-lactamase substrate, an AmpC inhibitor, and a serine beta-lactamase inhibitor in an amount sufficient to inhibit an ESBL and an OSBL but not a class A serine carbapenemase; and
(c) a third composition comprising a detectable beta-lactamase substrate and a serine beta-lactamase inhibitor in an amount sufficient to inhibit an ESBL and an OSBL but not a class A serine carbapenemase.
5. The kit of claim 1 further comprising a third composition comprising a detectable beta-lactamase substrate and an ESBL inhibitor.
6. The kit of claim 3 further comprising a fifth composition comprising a detectable beta-lactamase substrate and an ESBL inhibitor.
7. The kit of claim 1 , 2 or 4 , wherein the detectable beta-lactamase substrate is nitrocefin, pyridinium-2-azo-p-dimethylaniline chromophore, (7R)-3-(((3-carboxy-4-nitrophenyl)thio)methyl)-8-oxo-7-(2-(thiophen-2-yl)acetamido)-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, or (7R)-7[2-aminothiazol-4-yl]-(z)-2-(1-carboxy-1-methylethoxyimino)acetamido)-3-(2,4-dinitrostyryl)-3-cephem-4-carboxylic acid trifluoroacetate, E-isomer.
8. The kit of claim 1 , 2 or 4 , wherein the AmpC inhibitor is cloxacillin, a salt form of cloxacillin, (2S,3S)-3-(((Z)-2-(((1,5-dihydroxy-4-oxo-1,4-dihydropyridin-2-yl)methoxy)imino)-2-(thiophen-2-yl)ethyl)amino)-2-methyl-4-oxoazetidine-1-sulfonic acid, boronic acid, or a boronic acid derivative.
9. The kit of claim 1 , 2 or 4 , wherein the serine beta-lactamase inhibitor is clavulanic acid, tazobactum, sulbactam, or a salt form thereof.
10. The kit of claim 5 or 6 , wherein the ESBL inhibitor is cefotaxime, ceftazidime, or a salt form thereof.
11. The kit of claim 2 , wherein the metal chelator is dipicolinic acid (DPC), diethyl dithiocarbamate (DEDTC), N,N,N′,N′-tetrakis-(2-pyridylmethyl)-ethylenediamine (TPEN), Ethylenediaminetetraacetic acid (EDTA), 2,3-dimercapto-1-propane sulfonic acid (DMPS) or 1,10-phenathroline.
12. The kit of claim 1 , 2 or 4 , wherein each composition comprises a lysis reagent.
13. The kit of claim 1 , 2 or 4 , wherein each composition comprises a lysis reagent and an agent that promotes the stabilization of the lysis reagent.
14. The kit of claim 1 , 2 or 4 , wherein each composition comprises a lysis reagent, an agent that promotes the stabilization of the lysis reagent and an agent that enhances the lysis of a bacterial cell by a lysis reagent.
15. The kit of claim 1 , 2 or 4 , wherein each composition comprises lysozyme and trehalose.
16. The kit of claim 1 , 2 or 4 , wherein each composition comprises lysozyme, trehalose and EDTA.
17. The kit of claim 1 , 2 or 4 , wherein each composition is dried and present on or in a different solid support.
18. The kit of claim 12 , wherein each composition is dried and present on or in a different solid support.
19. The kit of claim 13 , wherein each composition is dried and present on or in a different solid support.
20. The kit of claim 14 , wherein each composition is dried and present on or in a different solid support.