Microfluidic Device for Microorganism Detection via Lysis and Nucleic Acid Separation
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Solution Overview
Problem
Current techniques fail to provide a simple, specific, and sensitive method for detecting microorganisms, lacking in terms of specificity, sensitivity, and operational simplicity.
Innovation Solution
A microfluidic device with a lysis section for dissolving microorganisms and a separation section for isolating nucleic acids, using bacteriolytic agents and separation means like molecular size or centrifugation, followed by complex formation and detection using CRISPR-dCas9 or protein markers, and electrochemical or optical detection through nanopores.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If PCR is used for microorganism detection, then detection sensitivity and specificity are improved, but device complexity and operational complexity increase
Solution Approach 1:
The detection system is divided into distinct functional modules: a lysis section for microorganism lysis, a separation section for nucleic acid isolation, and a detection section for signal generation. This segmentation allows each module to perform its specific function with optimized components, reducing overall system complexity while maintaining high detection sensitivity through specialized processing in each section.
Solution Approach 2:
The invention introduces intermediate processing steps between sample input and final detection: lysis agents break down microorganism cells to release nucleic acids, and separation means isolate the nucleic acids from other cellular components. These intermediary steps simplify the detection process by preparing the sample in a standardized format that the detection section can process directly, avoiding the need for complex PCR amplification while maintaining sensitivity.
2Ease of operation
If immunological detection method is used, then operation simplicity is improved, but detection sensitivity deteriorates
Solution Approach 1:
The invention replaces the mechanical/chemical immunological binding process with a nucleic acid-based detection system. Instead of using antibodies that require complex incubation and washing steps, the system uses nucleic acid extraction followed by direct detection methods, eliminating the need for immunological reagents and simplifying the operational protocol while achieving comparable or superior sensitivity.
Solution Approach 2:
The detection approach shifts from detecting protein antigens (immunological method) to detecting nucleic acid biomarkers. This parameter change allows for simpler sample preparation since nucleic acids are more stable and easier to extract from lysed cells than to maintain antibody-antigen complexes, while the specific nucleic acid sequences provide high detection sensitivity through sequence-specific recognition.
3Ease of operation
If ATP assay method is used, then operational simplicity is improved, but detection specificity deteriorates
Solution Approach 1:
The invention applies local quality by using specific nucleic acid sequences as biomarkers for different microorganism types. Instead of a general metabolic activity measurement (ATP), the system targets specific genetic sequences that are unique to pathogenic microorganisms, allowing the same simple operational platform to achieve high specificity through sequence-specific detection probes or primers in the detection section.
Solution Approach 2:
The lysis and separation sections provide universal processing for all microorganism samples, handling diverse microbial types through standardized nucleic acid extraction. The detection section then provides specific identification by targeting specific nucleic acid sequences. This universal-s specific architecture maintains operational simplicity across different sample types while achieving high detection specificity for various pathogenic microorganisms.
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
Enables specific and sensitive detection of microorganisms, suitable for various fields including food safety, with improved operational simplicity and sensitivity.
Implementation Method 1
a lysis section for dissolving microorganisms and a separation section for separating out a nucleic acid from a lysate of microorganisms formed in the lysis section
Implementation Method 2
the separation section includes a separation means capable of separating out a nucleic acid contained in the lysate from other substances
Implementation Method 3
a separation means using a molecular size, a separation means using centrifugation, or a separation means using hydraulic filtration
Implementation Method 4
a complex forming section for forming a protein-nucleic acid complex from the nucleic acid separated in the separation section
Implementation Method 5
a detection section including a means for detecting the complex communicating with the complex forming section
Implementation Method 6
a detection section including a means for detecting the complex communicating with the complex forming section
Implementation Method 7
a detection section detects the complex using a nanopore
Data Source
Figure 1(a)~1(b)
Figure 2(a)~2(d)
Figure 3
AI summary
A microfluidic device 1 according to the present invention for detecting microorganisms in a sample includes a lysis section 2 for dissolving microorganisms in the sample; and a separation section 3 for separating out a nucleic acid from a lysate of microorganisms formed in the lysis section 2 communicating with the lysis section 2, in which the lysis section 2 includes a channel for holding a bacteriolytic agent for lysing microorganisms, and the separation section 3 includes a means capable of separating out a nucleic acid contained in the lysate from other substances.