Multiplex Nucleic Acid Detection for Gastrointestinal Pathogens
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Solution Overview
Problem
There is a need for an efficient and sensitive method to detect Salmonella, Shigella, and Campylobacter in samples, particularly for diagnostic and prognostic purposes in cases of bacterial gastroenteritis.
Innovation Solution
A multiplex method involving the use of specific amplification oligomers and detection probes to amplify and detect the presence of Salmonella, Shigella, C. jejuni, and C. coli in a sample through in vitro nucleic acid amplification reactions, such as PCR or RT-PCR.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If multiple separate detection methods are used for each pathogen, then detection accuracy for each pathogen is maintained, but detection time and procedural complexity increase significantly
Solution Approach 1:
The patent combines multiple pathogen detection assays into a single multiplex PCR reaction. Different primer sets targeting Salmonella, Shigella, and Campylobacter are simultaneously included in one reaction mixture, allowing parallel amplification of multiple pathogen-specific DNA sequences. This merging approach maintains individual pathogen detection accuracy while reducing total detection time and procedural steps.
Solution Approach 2:
The detection system is designed with universal components that can detect multiple pathogens simultaneously. A single reaction mixture contains universal buffers, polymerases, and nucleotides that support amplification of different pathogen targets, while pathogen-specific primers and probes provide selective detection. This multi-functional design enables one assay to replace multiple separate assays.
2Reliability
If multiple separate detection methods are used for each pathogen, then cross-contamination risk is reduced, but device complexity and resource requirements increase
Solution Approach 1:
The patent uses pathogen-specific oligonucleotide primers and fluorescent probes as intermediaries that provide selective binding to target DNA sequences. These intermediaries ensure that each pathogen is detected through specific molecular recognition events within the multiplex reaction, maintaining reliability by preventing non-specific amplification and cross-reactivity even though multiple pathogens are detected simultaneously.
3Reliability
If traditional culture methods are used, then pathogen identification is confirmed, but detection sensitivity and time to result are insufficient
Solution Approach 1:
The patent performs preliminary amplification of pathogen-specific DNA sequences using PCR before final detection. This preliminary action enriches trace amounts of pathogen DNA in the sample, significantly increasing detection sensitivity. The amplified products then serve as templates for specific detection using fluorescent probes, maintaining reliable pathogen identification while enabling detection of much lower pathogen loads than traditional culture methods.
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
This method allows for the simultaneous detection of multiple bacterial pathogens, providing rapid and accurate diagnostic information that can aid in the treatment of bacterial gastroenteritis.
Implementation Method 1
specific amplification oligomers and detection probes to amplify and detect the presence of Salmonella, Shigella, C. jejuni, and C. coli in a sample through in vitro nucleic acid amplification reactions
Implementation Method 2
PCR or RT-PCR
Data Source
AI summary
Disclosed are nucleic acid oligomers, including amplification oligomers, detection probes, and combinations thereof, for detection of one or more gastrointestinal pathogens selected from Salmonella, Shigella, Campylobacter jejuni, and Campylobacter coli. Also disclosed are methods of specific nucleic acid amplification and detection, including multiplex assays, using the disclosed oligomers, as well as corresponding reaction mixtures and kits.