Flow Cytometry Microorganism Detection Resin Matrix Removal
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Solution Overview
Problem
Current methods for detecting microorganisms in complex matrices, such as food and environmental samples, face challenges including interference from matrix components, low detection sensitivity, and the need for time-consuming enrichment and selective plating, which limits the ability to accurately enumerate and identify pathogens in a timely manner.
Innovation Solution
The method involves using resins to modulate interference from complex matrices, followed by filtration to remove resin and matrix components, and then employing flow cytometry with fluorescent dyes and quenching agents to enhance signal detection and differentiate target microorganisms from background noise, allowing for rapid and accurate detection of microorganisms in complex samples.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If selective and differential plating is employed to identify specific microorganisms, then identification accuracy is improved, but detection time increases significantly
Solution Approach 1:
The patent extracts the identification function from the time-consuming selective and differential plating steps by using flow cytometry with fluorescently labeled antibodies that specifically bind to target microorganisms, allowing direct identification without culture enrichment
Solution Approach 2:
The patent replaces the mechanical culture and plating process with flow cytometry, an optical detection system that uses fluorescently labeled antibodies to identify microorganisms directly in the sample without requiring their cultivation
2Measurement precision
If enrichment steps are performed to increase target microorganism numbers, then detection sensitivity is improved, but assay time increases
Solution Approach 1:
The patent extracts the detection function from the enrichment process by using flow cytometry to directly detect and count target microorganisms in the original sample without requiring prior culture enrichment to increase their numbers
Solution Approach 2:
The patent introduces fluorescently labeled antibodies as intermediaries that specifically bind to target microorganisms, enabling direct detection and counting without the need for enrichment steps to amplify the target population
3Measurement precision
If centrifugation and filtration steps are used for sample preparation, then microorganism separation is improved, but microorganism viability is reduced
Solution Approach 1:
The patent replaces harsh mechanical separation methods like centrifugation and filtration with a gentle chemical approach using paramagnetic beads coated with affinity ligands that selectively capture target microorganisms through specific binding, minimizing mechanical stress and preserving viability
4Speed
If flow cytometry is used for rapid detection, then detection speed is improved, but detection sensitivity in complex matrices decreases
Solution Approach 1:
The patent introduces paramagnetic beads coated with affinity ligands as intermediaries that selectively capture target microorganisms from complex matrices, concentrating them and removing interfering substances before flow cytometry analysis, thereby enhancing detection sensitivity without sacrificing speed
Solution Approach 2:
The patent uses paramagnetic beads with porous structures that provide large surface areas for coating affinity ligands, enabling efficient capture and concentration of target microorganisms while allowing the removal of interfering matrix components
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 approach improves the sensitivity and accuracy of microorganism detection, reduces the need for sample enrichment, and enables the analysis of small sample sizes, providing a universal, rapid, and cost-effective method for detecting multiple microorganism strains in complex matrices without adverse effects on viability.
Implementation Method 1
utilizing both sample enrichment and selective plating results is a time-consuming assay... The resin is removed from the sample. The prepared sample is then combined with labels for the assay target
Implementation Method 2
employing flow cytometry with fluorescent dyes and quenching agents to enhance signal detection
Implementation Method 3
employing flow cytometry with fluorescent dyes and quenching agents to enhance signal detection
Implementation Method 4
employing flow cytometry with fluorescent dyes and quenching agents to enhance signal detection
Data Source
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AI summary
A process for determining one of the presence, absence, or total of microorganisms (e.g. bacteria) in a sample. According to the process, a biological sample containing complex matrices is obtained. The sample is first combined with a resin to adsorb complex matrices from the sample. The resin is removed from the biological sample. The sample so prepared is then analyzed by flow cytometry.