Multiplex Primer Assays for Rapid Urinary Microorganism Detection
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Solution Overview
Problem
Current diagnostic methodologies for urinary tract infections (UTIs) suffer from low throughput, lack of sensitivity and specificity, and are time-consuming, often missing pathogen detection in polymicrobial environments, and are costly and complicated.
Innovation Solution
A method involving multiple amplification reaction mixes with specific primer pairs targeting various nucleic acid sequences, followed by amplification and detection on a reaction vessel, utilizing an amplification product detection system to identify and profile urinary microbiota.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional culture-based methods are used for UTI diagnosis, then the workflow is simple and cost-effective, but the throughput is low, sensitivity and specificity are insufficient, and detection time is lengthy
Solution Approach 1:
The patent segments the detection process into multiple parallel amplification reactions, each targeting specific uropathogen nucleic acid sequences. By dividing the sample into multiple reaction mixes with different primer pairs, the system can simultaneously detect multiple pathogens, thereby increasing throughput while maintaining high sensitivity and specificity through targeted amplification.
Solution Approach 2:
The patent replaces the mechanical culture-based detection system with a molecular amplification system. Instead of relying on bacterial growth in culture media, the invention uses nucleic acid amplification techniques to directly detect and identify uropathogens, dramatically improving detection speed, sensitivity, and throughput while eliminating the need for lengthy incubation periods.
2Measurement precision
If traditional culture-based methods are used for UTI diagnosis, then the cost is lower, but the detection accuracy and pathogen identification capability are insufficient
Solution Approach 1:
The patent creates a universal detection platform that can identify multiple uropathogens using a common amplification and detection system. The same basic workflow and equipment can detect different pathogens by simply changing the primer pairs, making the system multi-functional. This universality helps reduce overall diagnostic costs compared to having separate specialized tests for each pathogen.
Solution Approach 2:
The patent combines multiple detection capabilities into a single integrated system. By merging the detection of multiple uropathogens into one amplification reaction mix or parallel reaction set, the system achieves high detection accuracy while reducing the need for multiple separate tests, thereby lowering overall diagnostic costs.
3Adaptability or versatility
If culture-based methods are used, then the workflow is straightforward, but the ability to detect polymicrobial infections is poor
Solution Approach 1:
The patent segments the detection capability across multiple primer pairs, each designed to target specific uropathogen sequences. This segmentation allows the system to simultaneously detect multiple different pathogens in a single sample, providing excellent polymicrobial infection detection capability. The segmented approach maintains manageable complexity by using modular primer pair designs.
Solution Approach 2:
The patent employs multiple primer pairs targeting different regions and pathogens, using an excessive number of targets beyond what a single culture method could detect. This partial detection of multiple potential pathogens simultaneously ensures that polymicrobial infections are comprehensively identified, with the added capability providing robust detection without overwhelming complexity.
4Speed
If traditional UTI diagnosis methods are used, then the process is simple, but the detection time is too long
Solution Approach 1:
The patent replaces the slow mechanical culture process with rapid nucleic acid amplification. By substituting the time-consuming bacterial growth mechanism with molecular amplification, the system achieves fast detection within hours rather than days. The increased complexity of the amplification system is justified by the dramatic improvement in detection speed.
Solution Approach 2:
The patent implements continuous amplification reactions that proceed without interruption to generate detectable products. The amplification process runs continuously through multiple cycles, maintaining useful action throughout the detection period, which significantly reduces total detection time compared to intermittent culture methods that require incubation, observation, and repeated sampling.
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
Provides rapid, accurate, and cost-effective detection and profiling of uropathogens in urine samples, overcoming limitations of traditional culture-based methods.
Implementation Method 1
forming at least five amplification reaction mixes each comprising an aliquot from a sample source comprising a plurality of nucleic acid sequences, using at least five different assays each comprising a pair of amplification primers; performing a plurality of amplification reactions on the reaction vessel; and detecting an amplification product corresponding to a target nucleic acid sequence
Implementation Method 2
each assay comprises a pair of amplification primers and a detectably labeled probe, the probe configured to specifically hybridize to the amplification product generated by the amplification primers
Data Source
AI summary
Various methods are disclosed for amplifying nucleic acid sequences in a nucleic acid sample. The methods involve forming at least five amplification reaction mixes each including an aliquot from a sample source that includes nucleic acid sequences, using at least five different assays each including a pair of amplification primers, the assays selected from the group of assays in Table 1 and/or targeting the sequences specified in Table 1.