Method and device for the detection of carbapenemases and carbapenemase producers

A method for the phenotypic detection of carbapenemase and carbapenemase producers, the method comprising:

• in a medium contacting a substrate of structural formula (I) with a sample suspected of containing the carbapenemases and/or carbapenemase producers:
• wherein
• R1 is absent or selected from the group consisting of —CH2—, —O—, —S—, —CO—, —CHOH—, and (—CH═CH)n—CH2—;
• R2 is selected from the group consisting of —H, —OH, —SH, halogen, a C1-C6 alkyl optionally substituted with —OH, ═O, —NH2, ═S, or —SH, an optionally substituted oxyalkyl, an optionally substituted alkenyl, an optionally substituted aromatic compound, an optionally substituted heterocyclic compound, a primary amine, a secondary amine, an ester, an ether, thiol, and a thioether;
• R is selected from the group consisting of a C1-C6 alkyl optionally substituted with —OH, ═O, —NH2, ═S, or —SH, an optionally substituted oxyalkyl, an optionally substituted alkenyl, an optionally substituted aromatic compound, an optionally substituted heterocyclic compound, a secondary amine, an ester, an ether, and a thioether;
• n is an integer from 1 to 4;
• (L) is an optional linker wherein
• i) (L) is selected from the group consisting of a C1-C12 alkyl, C1-C12 oxyalkyl, C1-C12 alkene, and C1-C12 alkyne, each optionally substituted by —OH, —SH, ═O, —NH2, ═S, halogen, a primary amine, a second amine, an ester, an ether and a thiol;
• or
• ii) (L) is selected from the group consisting of a thioether, an aromatic compound, a cyclic compound, an ester, an ether, a carbamate, and an amide;
• (X) is an optional group, wherein
• i) (X) is selected from the group consisting of a —S—R—, a sulphonate, a substituted tosylate, a mesylate, a thiophenolate, a substituted thiophenolate, —S-aryl-, a phenolate, a substituted phenolate, and —O-aryl-;
• or
• ii) (X) comprises a carbamate, a carbonate, a thiocarbamate, a thiocarbonate, an inorganic ester, an organic ester, an anhydride, an alcohol, a carboxylic acid, a halide, and diazo;
• or
• iii) (X) is —S—, —O—SO2), or —O—SO2—R—;
• A and Z form a fluorophore/quencher pair, or A represents a reporter group and Z represents a solid phase, or A represents a solid phase and Z represents a reporter group,
• wherein when A and Z form a fluorophore/quencher pair, then
• A is a quencher and Z is a fluorophore, or
• A is a fluorophore and Z is a quencher,
• and wherein
• if (X) is present, then the group X—Z is released after enzymatic cleavage of the substrate by carbapenemase, or
• if (X) is absent, then the group Z is released after enzymatic cleavage of the
• substrate by carbapenemase;
• and
• detecting the change in fluorescence intensity observed after the separation of the fluorophore from the quencher following cleavage of the substrate by the carbapenemase;
• or
• detecting the reporter group cleaved from or with the substrate following cleavage of the substrate by the carbapenemase.