Methods for the detection and identification of extended spectrum beta lactamases
AssigneeGeneOhm Sciences, Inc.
InventorsCatherine Lippe, Isabelle Duteaud, Celine Roger-Dalbert
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Abstract
Embodiments disclosed herein relate to compositions for the detection and/or identification of microbes that carry extended spectrum beta-lactamase genes. Specifically, provided herein are oligonucleotides, probes, and kits containing the same, for the detection of bacterial CTX-M sequences. Also provided are methods for the detection and/or amplification of microbes harboring extended spectrum beta-lactamase genes, including CTX-M type extended spectrum beta-lactamase genes.
Claims27
A method for detecting and or identifying of the presence of microbes with a CTX-M extended spectrum β-lactamase in a specimen, comprising:
- obtaining a sample from the specimen to be analyzed for the presence of a CTX-M gene;
- contacting the sample with a set of amplification primers under standard PCR conditions, wherein the set of amplification primers comprises at least one primer pair, wherein said set of primers comprises one or more primers with a universal base, wherein said primer pair hybridizes to nucleic acids flanking a target sequence within a CTX-M extended spectrum β-lactamase gene, and wherein said primer pair generates a target amplification product;
- providing reagents and conditions for extension of the primers to generate the target amplification product;
- and
- determining the presence and/or amount of the target amplification product, wherein the presence of the target amplification product indicate the presence of said CTX-M gene.
The method of claim 5, wherein said plurality of amplification primer pairs collectively hybridize to, and produce target amplification products in the presence of, the nucleic acids of CTX-M genes from CTX-M-1, CTX-M-2, CTX-M-8, CTX-M-9, and CTX-M-25 groups.
A kit for the detection and/or identification of the presence of microbes with CTX-M extended spectrum β-lactamases in a specimen, comprising:
- a set of amplification primers wherein the set of amplification primers comprises at least one primer pair, and wherein said set of primers comprises one or more primers with a universal base, wherein said at least one primer pair hybridizes to nucleic acids flanking a target sequence within or including part of a CTX-M extended spectrum β-lactamase gene.
The kit of claim 23, wherein said probe is selected from the group consisting of a molecular beacon probe, a TAQMAN™ fluorescent probe, and a scorpion fluorescent probe.
Dependencies are inferred from the claim text.