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Rapid nucleic acid assay for detecting CTX-M type ESBL-producing antibiotic resistant bacteria

Technology Benefits
Status: Technology Readiness Level (TRL) 3The assayΓÇÖs performance characteristics have beenpreliminarily assessed using ESBL-producing bacteriaextracted from patient samples and belonging to themost prevalent phylogenetic CTX-M subgroups. Theanalytical sensitivity of the assay is ~10 genomecopies per reaction. Verifying the excellenttheoretical coverage of the different CTX-Msubgroups with larger panels of strains is required.
Technology Application
The short amplicons and limited number of probes of thecurrent invention facilitate the integration of the assay inautomated nucleic acid testing platforms while allowingexceptional coverage, specificity and sensitivity of detection forthe various CTX-M gene variants.
Detailed Technology Description
The invention encloses a rapid closed-tube real-time nucleicacid assay for the detection of CTX-M variants:- High sensitivity and strain coverage using a single amplicon and a limited number of probes- Simultaneous detection of >90% of known CTX-M variants- Klebsiella oxytoca K1 gene classified as negative- Some embodiments enclose novel two-part chelate complementation probes allowing designing fully complementary probes for targets spotted with polymorphism.
*Abstract
Increasing numbers of bacteria are gaining ability to produce extended-spectrum ╬▓-lactamase (ESBL) enzymes which degrade extended-spectrum cephalosporin antibiotics.The most prevalent sub-class is CTX-M with more than 100 blaCTX-M gene variants encountered.Designing rapid and direct nucleic acid based detection methods for CTX-M with sufficient strain coverage and specificity has proven very problematic and typically require a battery of amplicons combined with DNA sequencing analysis.
Country/Region
USA

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