Bacterial Detection via Filter Membrane Binding and Lysis
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for detecting gram-positive and gram-negative bacteria in biological samples are slow, often requiring pre-culturing and are hindered by the presence of interfering substances, making them impractical for rapid detection in non-laboratory environments, especially when dealing with low numbers of bacteria.
Innovation Solution
The method involves filtration and treatment of the sample with non-ionic surfactants, ATP eliminating enzymes, and microbial lysing agents followed by the use of Luciferin/Luciferase reagents for luminescence-based quantitation, allowing for the rapid detection of bacteria without the need for pre-culturing and effectively addressing the interference from somatic cells and matrix substances.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If pre-culturing techniques are used to detect low numbers of bacteria, then detection sensitivity is improved, but detection time increases significantly (18-72 hours)
Solution Approach 1:
The patent applies preliminary action by pre-coating the filter membrane with specific binding agents (antibodies, lectins, or lytic enzyme binding domains) before sample application. This preliminary preparation enables immediate capture and concentration of target bacteria as they pass through the filter, eliminating the need for time-consuming pre-culturing steps while maintaining detection sensitivity for low bacterial concentrations (10^1-10^3 organisms/mL).
Solution Approach 2:
The patent extracts and concentrates bacteria from large volumes of biological sample through filtration onto a specialized membrane. The binding agents on the membrane selectively capture target bacteria, separating them from interfering substances in the sample matrix. This extraction and concentration process achieves detection sensitivity comparable to culturing methods but without requiring bacterial growth time.
2Measurement precision
If differential bacterial plate culturing is used to determine gram status, then identification accuracy is improved, but additional time is required (24-36 hours)
Solution Approach 1:
The patent uses an intermediary chemical treatment step involving selective lysing agents that differentiate between gram-positive and gram-negative bacteria based on their cell wall structure. After filtration and capture, specific chemical reagents selectively lyse one gram type while preserving the other, allowing gram status determination without prolonged culturing. This intermediary chemical differentiation provides rapid gram type identification with high accuracy.
3Measurement precision
If filtration and binding technologies are used to concentrate bacteria, then detection sensitivity for low numbers is improved, but device complexity increases
Solution Approach 1:
The patent merges multiple functions into a single integrated filter membrane structure. The membrane combines filtration capability with pre-coated binding agents for bacterial capture and concentration. This unified approach eliminates the need for separate filtration and concentration steps, reducing overall system complexity while maintaining high detection sensitivity for low bacterial numbers in the sample.
Solution Approach 2:
The patent employs a porous filter membrane as the core component, which provides both physical filtration and a large surface area for binding agent attachment. The porous structure allows efficient sample passage while capturing bacteria through the binding agents coated on the pore surfaces. This use of porous material achieves effective bacterial concentration without requiring complex mechanical concentration devices.
4Reliability
If sample enrichment is performed to detect low numbers of organisms, then detection reliability is improved, but productivity decreases due to extended processing time
Solution Approach 1:
The patent replaces the biological enrichment process (bacterial culturing) with a physical-chemical concentration system. Filtration and binding technologies physically concentrate bacteria from large sample volumes onto a small membrane surface, achieving the same enrichment effect as culturing but without requiring bacterial growth time. This substitution of mechanical/chemical methods for biological processes maintains detection reliability while dramatically improving productivity and enabling rapid results.
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 enables rapid, sensitive, and specific detection of bacteria, including gram status, in various biological samples, reducing detection time and improving the reliability of results, even in samples with low bacterial concentrations.
Implementation Method 1
Treatment of the filtrate with a non-ionic surfactant which lyses non-microbial cells (somatic cells)
Implementation Method 2
Treatment of the first solution with an ATP eliminating enzyme to produce a second solution
Implementation Method 3
Treatment of the third solution with a microbial lysing agent to give a fourth solution
Implementation Method 4
Treatment of the fourth solution with a Luciferin/Luciferase reagent to give a fifth solution and Quantitation of bacteria in the fifth solution by luminescence
Data Source
AI summary
The present invention relates methods for rapidly detecting the presence of bacteria in biological solutions regardless of their origin in non-laboratory environments. The present invention further relates to methods for binding, capturing, and concentrating the bacteria in a given sample.