CTX-M Detection Primers and Probes for Rapid Antibiotic Resistance Identification
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Solution Overview
Problem
There is a clinical need for rapid detection of antibiotic-resistant beta-lactamase genes, particularly CTX-M groups 1 and 9, which are prevalent in gram-negative bacteria, as they pose a significant challenge due to their role in conferring resistance to beta-lactam antibiotics.
Innovation Solution
The development of specific primers and probes that target the genes encoding extended-spectrum beta-lactamases, allowing for the amplification and detection of CTX-M groups 1 and 9 in biological samples using methods such as fluorescence resonance energy transfer (FRET) and hybridization detection probes, enabling sensitive and rapid identification of these resistance markers.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Speed
If conventional detection methods are used for antibiotic resistant bacteria, then detection can be performed, but detection speed and sensitivity are insufficient for rapid clinical intervention
Solution Approach 1:
The detection system is segmented into multiple functional components: specific primers for target recognition, probes for signal generation, and fluorescence detection systems for readout. This segmentation allows optimization of each component for speed and sensitivity independently, resolving the contradiction between rapid detection and high sensitivity requirements
Solution Approach 2:
The invention changes the detection parameter from conventional non-fluorescent methods to fluorescence-based detection. By using fluorophore-labeled probes and FRET (fluorescence resonance energy transfer) mechanisms, the system achieves both rapid signal acquisition and high sensitivity through optical parameter changes, simultaneously improving speed and detection precision
2Adaptability or versatility
If broad detection methods are used to detect all beta-lactamase genes, then all resistance types can be detected, but specific detection of clinically relevant CTX-M groups 1 and 9 becomes difficult
Solution Approach 1:
The primers and probes are designed with local quality optimization by targeting specific conserved regions within CTX-M groups 1 and 9 sequences. The primers contain nucleotide sequences that are highly specific to these clinically relevant groups, allowing differentiated detection while maintaining the ability to detect variations within these groups through careful sequence selection and hybridization condition optimization
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The described method allows for the specific, sensitive, and rapid detection of CTX-M groups 1 and 9 gene sequences, facilitating timely intervention in infections caused by antibiotic-resistant bacteria.
Implementation Method 1
The amplified nucleic acid can be detected by a variety of state of the art methods, including fluorescence resonance energy transfer (FRET)
Implementation Method 2
The amplified nucleic acids can also be detected by any combination of detection techniques which may include hybridization detection probes
Data Source
AI summary
Primers and probes specific to the genes encoding extended spectrum beta-lactamase that involves CTX-M groups 1 and 9 that cause extended beta-lactamase resistance in bacteria are described herein, with methods and kits for using these primers and probes to detect CTX-M groups 1 and 9 nucleic acids. In the methods described, nucleic acids present in a clinical or test sample obtained from a biological sample or tissue suspected of containing the CTX-M groups 1 and 9 gene are amplified and corresponding sequences for CTX-M groups 1 and 9 are detected. The amplified nucleic acid can be detected by a variety of state of the art methods, including fluorescence resonance energy transfer (FRET), radiolabels, enzyme labels, and the like.