Multiplex Nucleic Acid Amplification for Microbial Detection
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for monitoring and detecting vaginal, urogenital, and urinary tract microbial flora are costly, lack sensitivity and specificity, and require complex or lengthy workflows, necessitating the development of more efficient and cost-effective systems for assessing and profiling infection-causing microorganisms.
Innovation Solution
The use of methods and kits that perform parallel nucleic acid amplification reactions with specific primers and probes to detect and profile microorganisms, allowing for the simultaneous amplification and detection of multiple target sequences in a single sample, using amplification primer pairs and detectably labeled probes to identify specific microorganisms and biomarkers associated with infections.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If current technologies are used for vaginal and urinary microbial flora monitoring, then detection can be performed, but the methods are costly and require complicated or lengthy workflows
Solution Approach 1:
The diagnostic panel is segmented into multiple individual amplification reactions, each targeting specific microorganisms or microbial groups. Each reaction contains specific primers and probes designed for particular pathogens, allowing parallel processing of multiple targets in separate reaction chambers. This segmentation enables simplified, cost-effective individual assays while maintaining comprehensive detection capability through multiplexing.
Solution Approach 2:
The invention creates a universal detection platform that can monitor multiple microorganisms (bacteria, fungi, viruses, parasites) and microbial flora using a single integrated system. The same basic amplification reaction framework can detect diverse targets by simply changing the primer and probe sets, eliminating the need for separate specialized tests for each microorganism type and reducing overall workflow complexity.
2Measurement precision
If current technologies are used for microbial detection, then monitoring can be performed, but sensitivity and specificity are insufficient
Solution Approach 1:
The invention employs locally optimized primer and probe sequences tailored to specific microorganism targets. Each amplification reaction uses primers and probes with specific binding characteristics designed for their particular target, ensuring high specificity. The detection probes are designed with specific fluorophore-quencher configurations that provide localized signal generation only when the correct target is present, enhancing both sensitivity and specificity for each individual detection.
Solution Approach 2:
The invention uses fluorescently labeled probes that produce detectable color/fluorescence changes when they bind to their target nucleic acid sequences. The probe design incorporates fluorophores that emit specific wavelengths of light, allowing sensitive detection of amplified products. The fluorescence signal provides a quantifiable measure of detection sensitivity, while the specificity of fluorescence emission wavelengths allows simultaneous detection of multiple targets without cross-interference.
3Adaptability or versatility
If multiple microorganisms are detected simultaneously, then comprehensive profiling is achieved, but the workflow becomes more complex
Solution Approach 1:
The invention merges multiple individual amplification reactions into a single integrated diagnostic platform. Multiple microorganisms can be detected simultaneously in parallel reaction chambers using the same basic workflow, reagents, and detection system. The combining of multiple targets into one assay reduces workflow complexity compared to performing separate tests for each microorganism, while maintaining comprehensive profiling capability through multiplexed detection.
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
These methods enable sensitive, specific, and cost-effective monitoring and profiling of microbial flora, facilitating the identification of pathogenic microorganisms and imbalance in vaginal and urinary tract microbiota, which can inform treatment decisions and monitor treatment outcomes.
Implementation Method 1
performing a plurality of amplification reactions in parallel, at least one of the amplification reactions containing a portion of a nucleic acid sample and a pair of amplification primers configured to produce an amplification product
Implementation Method 2
at least one of the amplification reactions containing a detectably labeled probe configured to hybridize to a nucleic acid sequence generated by extension of at least one of the amplification primers
Implementation Method 3
performing a plurality of amplification reactions in parallel... forming a plurality of different amplification products
Data Source
AI summary
Methods, compositions and kits for detecting microorganisms and/or profiling microbiota such as for example through use of nucleic acid amplification and detection.