Micro-channel reactor for detecting drug-resistant brucella mateitae

Abstract

The invention provides a micro-channel reactor for detecting drug-resistant Maltese brucella. The micro-channel reactor comprises a micro-channel main body, a circulation micro-channel structure, a micro-sensor system, a control unit and a gene detection unit, a plurality of micro-channels are arranged in the micro-channel main body, the diameter of each micro-channel is 50-500 microns, and a feed port and a discharge port are respectively formed in two ends of the micro-channel main body; hydrophilic and hydrophobic patterns are arranged on the surface of the micro-channel and are realized by a method of combining photoetching and surface modification, and fluid is guided to flow along a preset path; the micro-channel is connected with a spiral circulation micro-channel structure, a nano catalytic structure layer is constructed on the surface of the inner wall of the circulation micro-channel structure, and a layer-by-layer self-assembly technology is adopted for preparation; the gene detection unit is a multi-mode sensor array and comprises a micro mass spectrum sensor, a fluorescence sensor, an electrochemical sensor and real-time PCR (Polymerase Chain Reaction) and / or CRISPR-Cas (Clustered Regularly Interspaced Short Palindromic Repeats). The detection efficiency is improved, closed pathogen detection and low-abundance sample enrichment are realized, and the technical effects of improving the detection sensitivity and the like are achieved.

Description

Technical Field

[0001] The present invention relates to the field of drug-resistant bacteria gene detection, and in particular to a microchannel reactor for drug-resistant Brucella Malta detection. Background Art

[0002] Brucellosis is a Class B infectious disease in my country. Its zoonotic nature allows drug-resistant strains to spread from animals (cattle and sheep) to humans, complicating epidemic control. Resistance rates among Maltese Brucella to first-line drugs (such as rifampicin and doxycycline) have been increasing annually. For example, resistance rates among Brucella melitensis in Egypt to ciprofloxacin and rifampicin reached 76.2%, while isolates from the Ulanqab region of Inner Mongolia in my country showed resistance to co-trimoxazole at 7%, with azithromycin resistance detected in 100% of isolates. Drug resistance can lead to failure of standard treatment regimens (such as doxycycline + rifampicin), prolonging the course of illness and increasing the risk of complications (such as liver damage and spondylitis). For example, the MIC of rifampicin-resistant strains can rise to 64 mg / L (compared to 1.0 mg / L for susceptible strains).

[0003] Current detection methods primarily include etiological testing, serological testing, and molecular biological testing. Etiological testing involves culturing blood / bone marrow samples using selective media (such as Brucella agar) combined with biochemical identification (catalase positive, oxidase positive, and weakly positive for urease). This culture takes 3-7 days and has a positive rate of only 60-80%, delaying treatment. This drug-resistant bacteria requires a high-security BSL-3 laboratory, making it difficult for primary care settings to conduct. Traditional antimicrobial susceptibility testing (broth dilution) takes an additional 2-3 days and lacks standardized interpretation guidelines from the CLSI. Serological testing methods, such as the tube agglutination test (SAT) and ELISA, measure antibody titers (IgM ≥ 1:160 for a positive result). These tests are prone to false positives and negatives, including cross-reactivity with Yersinia species O9 and Escherichia coli O157, and fluctuations in antibody titers following chronic infection or vaccination. They only reflect infection status and do not indicate strain resistance, making it impossible to distinguish between resistant strains.

[0004] With the rapid development of microfluidics and nanocatalysis technologies, microchannel reactors, a new type of reaction device, have been widely used in chemical synthesis, biomedicine, environmental management, and other fields due to their advantages such as high mass and heat transfer efficiency, controllable reaction conditions, and excellent safety. Microchannel reactors are typically composed of micron-scale channels, which enable precise control and efficient mixing of fluids, providing an ideal closed microenvironment for chemical reactions and are suitable for microbial detection with relatively high safety requirements.

[0005] However, the existing microchannel reactor technology still has some problems that need to be solved urgently: First, the catalytic efficiency of traditional microchannel reactors is low, mainly due to the uneven distribution of catalysts in the microchannels and insufficient effective catalytic surface area, resulting in long reaction time and low product yield; second, the existing microchannel reactors have limitations in fluid control, and it is difficult to accurately control the residence time of reactants in the catalytic zone, affecting the reaction efficiency and selectivity; third, the substrate damage problem often occurs during the hydrophilic-hydrophobic patterning modification process in the existing technology, affecting the service life and stability of the microchannel; fourth, the traditional microchannel reactor lacks real-time monitoring capabilities, and cannot adjust the reaction parameters in time, making it difficult to adapt to different reaction conditions and product requirements, resulting in a long product development cycle and low product yield; finally, the existing microchannel reactor system has deficiencies in integration and modularity, making it difficult to achieve multifunctional integration, especially in the field of medical applications. The existing bacterial resistance detection system has long sample processing time, complex operation, and low detection efficiency.

[0006] Although microchannel reactors (MICRs) have shown promise in bacterial resistance testing, they suffer from several drawbacks and limitations that hinder their application. For example, rapid analysis of large numbers of samples is essential for bacterial resistance testing, particularly in clinical and veterinary settings. However, MICRs suffer from low efficiency, a particularly acute problem during public health emergencies or large-scale epidemics. In practice, these devices require high technical expertise and specialized knowledge to operate correctly, increasing training and operating costs. Furthermore, the multi-step testing process further complicates operations and limits their widespread adoption in healthcare settings. Furthermore, the data generated by MICRs requires advanced data analysis techniques and tools to interpret. Due to their high throughput, the testing process generates large amounts of data, posing a significant challenge in effectively processing and analyzing this data. Manual processing alone, without a dedicated data processing team, can significantly impact the accuracy and reliability of test results. This is particularly problematic in resource-poor settings, where it can hinder the timeliness and accuracy of test results.

[0007] Therefore, there is an urgent need to develop a microchannel reactor for the detection of drug-resistant Brucella malta that can improve catalytic efficiency, control reaction rate and selectivity, avoid substrate damage, increase the lifespan and stability of the microchannel, and adapt to the needs of different applications. At the same time, for its application in bacterial resistance detection, there is still a need to improve detection efficiency and automation, helping medical institutions save time and costs and better serve patients. Summary of the Invention

[0008] In order to solve the technical problems that traditional culture of drug-resistant Brucella malta as a tertiary pathogen requires a high-level biosafety laboratory, the detection risk is high, and the traditional microchannel reactor has low catalytic efficiency, long reaction time, low detection efficiency, and is prone to environmental and personnel pollution, the present invention provides a microchannel reactor for the detection of drug-resistant Brucella malta, which can increase the catalytic efficiency by 3-5 times, shorten the reaction time by more than 60%, improve the detection efficiency, and realize the technical effects of closed pathogen detection, low-abundance sample enrichment, and improved detection sensitivity.

[0009] The present invention provides a microchannel reactor for detecting drug-resistant Brucella malta, comprising a microchannel body, a circulating microchannel structure, a microsensor system, a control unit, and a gene detection unit; the microchannel body is made of a polymer material, including one or more of polydimethylsiloxane (PDMS), polymethyl methacrylate (PMMA), polytetrafluoroethylene (PTFE), polycarbonate (PC), and cyclic olefin copolymer (COC); Multiple microchannels are provided inside the microchannel body, each with a diameter of 50~500μm, and a feed port and a discharge port are provided at both ends of the microchannel body respectively; the surface of the microchannel has a hydrophilic and hydrophobic pattern, which is achieved by a combination of photolithography and surface modification, guiding the fluid to flow along a preset path; the microchannel is connected to a spiral-shaped circulation microchannel structure, and a nanocatalytic structure layer is constructed on the inner wall surface of the circulation microchannel structure, which is prepared by layer-by-layer self-assembly technology; the microsensor system is integrated at a key position of the microchannel reactor, including one or more temperature sensors, pressure sensors, pH sensors and / or optical sensors; the control unit adjusts the working parameters and / or optimizes the reaction conditions according to the data feedback from the microsensor system; the genetic detection unit is a multimodal sensor array, including a micro mass spectrometer sensor, a fluorescence sensor, an electrochemical sensor, real-time PCR and / or CRISPR-Cas.

[0010] The microchannel reactor for detecting drug-resistant Brucella malta, provided by this invention, utilizes: 1) a nanocatalytic structure layer constructed on the inner wall of the microchannel to significantly increase the catalytic surface area; 2) hydrophilic and hydrophobic patterning technology to create specific fluid channels on the channel surface to control the fluid flow path; 3) a design of a circulating microchannel structure to extend the residence time of reactants in the catalytic zone; 4) an integrated microsensor for real-time monitoring of reaction parameters; 5) a control unit that automatically adjusts operating parameters and optimizes reaction conditions; and 6) an integrated genetic detection unit for online differentiation of drug resistance in drug-resistant Brucella malta. This design not only improves catalytic efficiency, controls reaction rate and selectivity, and addresses current issues such as low detection efficiency and long detection time, but also enables closed-loop detection of highly pathogenic bacteria. Furthermore, through microchannel pre-enrichment technology, trace samples (such as joint fluid) are concentrated above the detection threshold, improving sensitivity and integrating trace sample enrichment with safety testing.

[0011] Preferably, the microchannel reactor provided by the present invention is a microchannel surface that is scanned three-dimensionally controlled by a computer through a femtosecond laser system to create a complex micro-nano structure, which is combined with chemical modification to form a hydrophilic and hydrophobic pattern to control the flow path of the fluid; the circulation microchannel structure includes one or more micro-mixing units. The improved laser processing technology solves the problem of substrate damage in traditional methods. Traditional methods use nanosecond lasers or chemical etching, which easily cause thermal damage or chemical corrosion to the substrate, affecting the mechanical strength and service life of the microchannel. The improved femtosecond laser processing technology uses the principle of cold processing to concentrate energy and release it in an extremely short time, reducing heat diffusion and avoiding thermal damage to the substrate. At the same time, the laser processing process does not use chemical reagents, avoiding the impact of chemical corrosion on the substrate. The mechanical strength of the microchannel treated with the improved laser processing technology is 30% higher than that of the traditional method, and the service life is extended by 50%.

[0012] Preferably, the microchannel reactor provided by the present invention comprises a multilayer filtration structure within the microchannels, composed of membranes with different functions. These membranes are formed using a membrane substrate made from a blend of polysulfone (PSF) and polyethersulfone (PES) in a mass ratio of 7:3. The nanoreinforcers are carbon nanotubes and / or silica nanoparticles, prepared using a phase inversion method. Different pore sizes and chemical modifications are employed to enhance separation efficiency and selectivity. This novel membrane material exhibits excellent mechanical strength, chemical stability, and thermal stability. Its tensile strength is 50 MPa, twice that of conventional membrane materials. Its chemical resistance is stable within a pH range of 2-12. Its thermal stability is also significant, with a glass transition temperature of 220°C, 50°C higher than that of conventional membrane materials.

[0013] Preferably, the microchannel reactor provided by the present invention utilizes a deformable microchannel structure using piezoelectric materials and shape memory alloys, enabling the microchannel width to continuously vary within a range of 50-500 μm. The piezoelectric actuator is made of polyvinylidene fluoride (PVDF), and the shape memory alloy is a nickel-titanium alloy, distributed at key locations in the microchannel. When voltage is applied to the piezoelectric actuator, the piezoelectric material deforms, pushing the microchannel walls to deform. When the shape memory alloy is heated, the alloy returns to its preset shape, pulling the microchannel walls back to their original shape. The present invention utilizes a deformable microchannel structure, using piezoelectric materials or shape memory alloys to dynamically adjust the channel geometry (the channel width can be continuously varied within a range of 50-500 μm), optimizing reaction conditions and separation efficiency. For example, in the initial reaction phase, the channel width can be set to a larger value (e.g., 300 μm) to increase the flow rate of reactants; in the middle phase, the channel width can be reduced (e.g., 100 μm) to prolong the residence time of reactants in the catalytic zone; and in the later phases of the reaction, the channel width can be restored to a larger value to accelerate product discharge.

[0014] Preferably, the microchannel reactor provided by the present invention, wherein one or more microchannel reactors are integrated on a chip, comprises an injection area, a reaction area and a detection area; the microchannel reactor adopts a multi-layer structure design, comprising a reaction layer, a heating layer, a cooling layer and a sensing layer, the reaction layer comprises a microchannel body and a circulating microchannel structure, the heating layer comprises a micro heating element, the cooling layer comprises a micro Peltier element, and the sensing layer comprises one or more micro sensors.

[0015] Preferably, the microchannel reactor provided by the present invention, wherein the chip adopts a modular design, including a basic platform and functional modules; the basic platform includes a power supply system, a control system and a communication system, and the functional modules include a reaction module, a separation module, a detection module and / or a storage module; each module adopts a plug-and-play method and is replaced according to the separation or detection needs. The modules are connected through standard interfaces and adopt a plug-and-play method, without the need for complicated installation and debugging processes. For example, when different types of catalytic reactions need to be carried out, only the reaction module needs to be replaced, and the other modules remain unchanged; when different types of products need to be detected, only the detection module needs to be replaced, and the other modules remain unchanged. The modular design greatly improves the adaptability of the system, enabling it to cope with various complex reactions and separation tasks.

[0016] Preferably, the microchannel reactor provided by the present invention integrates intelligent control software and adopts a modular design, including a data acquisition module, a parameter control module, a data analysis module and / or a user interface module; the data acquisition module obtains the reaction data of each sensor in the microsensor system; the parameter control module feedback adjusts the operating parameters of the microchannel reactor, the data analysis module performs real-time analysis of the collected data, and the user interface module provides a user-friendly operation interface to display the reaction status and analysis results. An intelligent control algorithm is used, based on machine learning technology, to analyze the reaction parameters and separation effect in real time, and automatically adjust parameters such as microchannel shape, flow rate and temperature; the modular design allows for rapid replacement of microchannel reactors and separation units with different functions, thereby improving system adaptability. The intelligent control software can accurately manage the flow rate and temperature of multiple reaction units, with a flow rate control accuracy of ±0.01μL / min and a temperature control accuracy of ±0.1℃. Through the coordinated management of the intelligent control software, the sample processing time is shortened from several hours of traditional methods to less than 30 minutes, greatly improving work efficiency.

[0017] Preferably, the microchannel reactor provided by the present invention is used to detect bacterial resistance of drug-resistant Brucella malta, forming a bacterial resistance detection system. The bacterial resistance detection system includes a hardware layer, which includes a microfluidic chip integrated into the microchannel reactor and includes one or more functional areas: a sample pretreatment area, a bacterial culture area, an antibiotic gradient area, and a detection area. The detection area includes a gene detection unit, which is a multimodal sensor array including a micro-mass spectrometer sensor, a fluorescence sensor, an electrochemical sensor, a real-time PCR, and / or CRISPR-Cas, distributed at different locations in the microchannel reactor. The microchannel reactor for detecting drug-resistant Brucella malta shortens the bacterial resistance detection cycle from 6 hours to a week in traditional methods to 2-4 hours, improving detection efficiency several times. The integrated multimodal sensor array (including micro-mass spectrometer, fluorescence, and electrochemical sensors) comprehensively monitors the reaction progress and product quality. These sensors are distributed at different locations in the microchannel reactor to form a sensor network that comprehensively monitors the reaction progress and product quality. The multimodal sensor array provides rich reaction information, enabling the system to more comprehensively understand the reaction state and make more accurate control decisions. Through these optimization measures, the platform achieves more efficient drug resistance detection, shortening detection time by 60% and reducing the number of steps by 70% compared to traditional methods.

[0018] Preferably, in the microchannel reactor provided by the present invention, the bacterial resistance detection system further includes a control layer and a software layer, the control layer includes a microprocessor and / or a driving circuit, and the software layer adopts a microfluidic design, including channel structure optimization and / or material optimization, the channel structure optimization adopts a three-dimensional structure, and the material optimization adopts biocompatible materials.

[0019] This bacterial resistance detection system combines intelligent control software, microchannel reactors and microfluidic chip design to form an integrated medical application platform. The intelligent control software accurately manages the flow rate and temperature of multiple reaction units to achieve automatic sample processing and reaction control in a short time; the microfluidic chip further shortens the bacterial resistance detection cycle of drug-resistant Brucella maltese and improves detection efficiency by more than 50%; the online miniature mass spectrometer monitors sample processing and reaction progress in real time to ensure the accuracy and timeliness of data, making the entire system more intelligent and automated to meet the needs of high-throughput detection.

[0020] Preferably, the microchannel reactor provided by the present invention, wherein the reactants enter the microchannel through the feed port and flow along a preset path under the guidance of the hydrophilic and hydrophobic patterned surface of the microchannel; after entering the circulation microchannel structure, the reactants undergo a catalytic reaction under the action of the nano-catalytic structure layer, the reaction parameters are monitored in real time by the microsensor system, and the reaction conditions are adjusted by the control unit, and the reaction products finally flow out from the discharge port and are collected.

[0021] The beneficial effects of the present invention are: by constructing a nano-catalytic structure layer on the inner wall of the circulating microchannel structure, the catalytic efficiency is increased by 3-5 times, the reaction time is shortened by more than 60%, and the detection yield is greatly improved; by adopting hydrophilic-hydrophobic patterning technology and multi-layer filtration structure, the production cycle of the traditional batch process is shortened by 60-70%, and the solvent usage is reduced by 80-90%. Combined with intelligent control software and microfluidic chip technology, the bacterial resistance detection cycle is shortened and the detection efficiency is increased by several times; through deformable microchannel structure and intelligent control algorithm, the product development cycle is shortened, and the flexibility and efficiency of drug production and bacterial detection are enhanced; improving laser processing technology and adopting new membrane materials to solve the problem of substrate damage in hydrophilic-hydrophobic patterning modification, realize a more sturdy and durable multi-layer filtration structure, ensure integrated reaction, separation, and purification processes, realize real-time monitoring, improve system adaptability, adapt to the application needs of multiple fields, and ensure stability and reliability in long-term use. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for use in the description of the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0023] Figure 1 Schematic diagram of the bacterial resistance detection system for Brucella malta DETAILED DESCRIPTION

[0024] To further illustrate the present invention, examples are provided below. It should be noted that these examples are purely illustrative. These examples are provided to fully illustrate the significance and content of the present invention, but are not intended to limit the present invention to the scope of the embodiments. The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings. Example 1

[0025] A microchannel reactor for detecting drug-resistant Brucella malta includes a microchannel body, a hydrophilic and hydrophobic patterned surface, a nanocatalytic structure layer, a circulating microchannel structure, a microsensor system and a gene detection unit.

[0026] The microchannel body is made of polydimethylsiloxane (PDMS) and measures 50 mm in length, 10 mm in width, and 5 mm in height. It contains multiple parallel microchannels, each with a diameter of 200 μm. The microchannel body has an inlet and outlet at each end. The inlet is connected to a microsyringe pump to control the feed rate of the reactants, while the outlet is connected to a collection device for the reaction products.

[0027] Hydrophilic-hydrophobic patterning technology is applied to microchannel surfaces through a combination of photolithography and surface modification. Specifically, a photosensitive material is first coated on the microchannel surface, then irradiated with ultraviolet light through a mask to form a specific pattern. Next, the surface is chemically modified to introduce hydrophobic groups (such as perfluoroalkylsilane) and hydrophilic groups (such as hydroxyl or amino groups) into specific areas. This hydrophilic-hydrophobic patterned surface can precisely control the flow path of the fluid, forming a stable liquid microchannel, avoiding disordered diffusion of reactants, and improving reaction selectivity. The contact angle of the hydrophilic area is less than 30°, while the contact angle of the hydrophobic area is greater than 150°, creating a significant surface energy difference that effectively guides the fluid along the preset path.

[0028] The microchannels are connected to a circular flow microchannel structure. The circular flow microchannel structure is designed in a spiral shape with a total length of 200 mm, forming multiple annular flow paths within the microchannel body. The circular flow microchannel structure includes a main loop and multiple branch loops, with the main loop having a diameter of 2 mm and the branch loop having a diameter of 0.5 mm. The design of the circular flow structure extends the residence time of the reactants in the catalytic zone to 5-10 times that of traditional straight channels, thereby improving the reaction conversion rate. The circular flow microchannel structure also features multiple micro-mixing units that utilize a passive mixing principle and enhance fluid mixing through special geometric structures (such as herringbone and zigzag shapes), thereby improving mass transfer efficiency.

[0029] The nanocatalytic structural layer is constructed on the inner wall surface of the circulating microchannel and is prepared using layer-by-layer self-assembly technology. First, the inner wall surface of the circulating microchannel is plasma treated to give the surface a negative charge; then, it is alternately immersed in a polyelectrolyte solution and a metal nanoparticle solution to form a multilayer composite structure. The thickness of the nanocatalytic structural layer is 50-200nm and it is composed of gold nanoparticles, palladium nanoparticles, and platinum nanoparticles with particle sizes of 5nm, 8nm, and 10nm, respectively. This nanocatalytic structural layer has highly dispersed active sites and a large specific surface area, which is about 200 times larger than the surface area of ​​traditional planar catalysts, greatly improving the catalytic efficiency.

[0030] The microsensor system is integrated into key locations within the microchannel reactor and includes temperature, pressure, pH, and optical sensors. The temperature sensor uses a microthermocouple with a measurement range of -50°C to 300°C and an accuracy of ±0.1°C. The pressure sensor uses a piezoresistive sensor with a measurement range of 0-10 MPa and an accuracy of ±0.01 MPa. The pH sensor uses a micro ion-selective field-effect transistor with a measurement range of 0-14 and an accuracy of ±0.01. The optical sensor uses a micro fiber optic spectrometer with a wavelength range of 200-800 nm and a resolution of 0.5 nm. These sensors are connected to the control unit via a micro data acquisition system, enabling real-time monitoring and feedback control of reaction parameters.

[0031] The gene detection unit is a multimodal sensor array, including a micro mass spectrometry sensor, a fluorescence sensor, an electrochemical sensor, real-time PCR and / or CRISPR-Cas.

[0032] The microchannel reactor is also equipped with a temperature control system, including micro-heating elements and cooling units, which can precisely control the reaction temperature within a range of -20°C to 200°C, with temperature fluctuations within ±0.5°C. The heating elements are micro-resistance heating wires evenly distributed around the microchannels, while the cooling units use micro-Peltier elements for rapid temperature reduction.

[0033] In practical applications, reactants enter the microchannel through the feed port and flow along a pre-set path, guided by the hydrophilic and hydrophobic patterned surface. Once inside the circulation microchannel structure, the reactants undergo a catalytic reaction under the action of the nanocatalytic structure layer, while a microsensor system monitors the reaction parameters in real time. The reaction products ultimately flow out of the discharge port and are collected. Throughout this process, the control unit automatically adjusts parameters such as the feed rate and reaction temperature based on sensor feedback to optimize reaction and detection conditions. Example 2

[0034] A microchannel reactor for the detection of drug-resistant Brucella malta can further solve the problem of substrate damage in hydrophilic and hydrophobic patterning modification by improving laser processing technology, and achieve a more robust and durable multi-layer filtration structure by adopting new membrane materials.

[0035] The improved laser processing technology uses a femtosecond laser system with a laser wavelength of 800nm, a pulse width of 100fs, a repetition rate of 1kHz, and an energy density of 0.5-2J / cm². Compared with traditional nanosecond lasers, femtosecond lasers have shorter pulse widths and more precise energy control, enabling fine processing on microchannel surfaces without causing thermal damage. During the laser processing process, a computer-controlled three-dimensional scanning system is used with a scanning speed of 10mm / s and a scanning accuracy of 1μm, which can create complex micro-nanostructures on the microchannel surface. These micro-nanostructures include micropillar arrays, microgrooves, and micropores, with sizes ranging from 1-50μm and depths ranging from 0.5-10μm. These micro-nanostructures, combined with subsequent chemical modification, form precise hydrophilic and hydrophobic patterns to control the flow path of the fluid.

[0036] Improved laser processing technology solves the substrate damage problem encountered by traditional methods. Traditional methods use nanosecond lasers or chemical etching, which can easily cause thermal damage or chemical corrosion to the substrate, affecting the mechanical strength and service life of the microchannels. Improved femtosecond laser processing technology utilizes the principle of cold processing, concentrating energy release within a very short period of time, reducing heat diffusion and avoiding thermal damage to the substrate. Furthermore, the laser processing process does not use chemical reagents, avoiding the effects of chemical corrosion on the substrate. Microchannels processed using this improved laser processing technology have a 30% increase in mechanical strength and a 50% increase in service life compared to traditional methods.

[0037] The microchannels contain a multi-layered filtration structure composed of membranes with different functionalities. The new membrane material utilizes a blend of polysulfone (PSF) and polyethersulfone (PES) with the addition of a crosslinker and nanoreinforcers. The mass ratio of PSF to PES is 7:3. The crosslinker is epoxy resin, added at 5%, while the nanoreinforcers are carbon nanotubes and silica nanoparticles, added at 1% and 2%, respectively. This new membrane material exhibits excellent mechanical strength, chemical stability, and thermal stability. Its tensile strength is 50 MPa, twice that of conventional membrane materials. Its chemical resistance is stable within a pH range of 2-12. Its thermal stability is 50°C, with a glass transition temperature of 220°C, 50°C higher than that of conventional membrane materials.

[0038] The new membrane material is prepared using a phase inversion method. First, polysulfone and polyethersulfone are dissolved in N,N-dimethylformamide (DMF) to form a 15% polymer solution. A crosslinker and nanoreinforcement agent are then added, and ultrasonic dispersion is performed for 30 minutes. The mixed solution is then cast onto a glass plate to a controlled thickness of 200 μm. Finally, the cast membrane is immersed in deionized water for phase inversion, forming a porous membrane structure. The prepared membrane undergoes heat treatment (120°C for 2 hours) and surface modification to create a multilayer filtration structure with specific pore sizes and surface properties.

[0039] The multi-layer filtration structure consists of three layers of membranes with different functions. From top to bottom, they are: the first layer is a macroporous membrane with a pore size of 1μm and a thickness of 200μm, which is mainly used to filter large particles of impurities; the second layer is a mesoporous membrane with a pore size of 0.2μm and a thickness of 150μm, which is mainly used to filter microorganisms and macromolecules; the third layer is a microporous membrane with a pore size of 50nm and a thickness of 100μm, which is mainly used to filter nanoparticles and small molecules. This multi-layer filtration structure can effectively separate substances of different sizes, improving separation efficiency and selectivity.

[0040] Improved laser processing technology, new membrane materials, and a multi-layer filtration structure have resulted in microchannel reactors with enhanced durability and stability. In tests conducted after 1,000 hours of continuous operation, the filtration efficiency of the new membrane material decreased by no more than 5%, while that of conventional membrane materials decreased by more than 30%. The mechanical strength of the new membrane material also decreased by no more than 10%, while that of conventional membrane materials decreased by more than 40%. This high durability and stability enables the microchannel reactor to operate stably and long-term under harsh conditions, significantly extending the equipment's service life and reducing maintenance costs. Example 3

[0041] A microchannel reactor for detecting drug-resistant Brucella malta can further adopt a deformable microchannel structure to optimize reaction conditions and separation efficiency.

[0042] The deformable microchannel structure is achieved by combining piezoelectric materials and shape memory alloys. The piezoelectric material is polyvinylidene fluoride (PVDF), 50 μm thick, coated with gold electrodes on both sides to form a piezoelectric actuator. The shape memory alloy is nickel-titanium alloy (Nitinol), 100 μm in diameter, and is distributed at key locations in the microchannel. When voltage is applied to the piezoelectric actuator, the piezoelectric material deforms, pushing the microchannel walls to deform. When the shape memory alloy is heated, it returns to its preset shape, pulling the microchannel walls to deform. By controlling the piezoelectric actuator voltage (0-100 V) and the shape memory alloy temperature (30-80°C), the microchannel width can be continuously varied within a range of 50-500 μm. This deformable microchannel structure dynamically adjusts the channel geometry according to reaction requirements, optimizing reaction conditions and separation efficiency. For example, in the early stage of the reaction, the channel width can be set to a larger value (such as 300 μm) to increase the circulation rate of the reactants; in the middle stage of the reaction, the channel width can be reduced (such as 100 μm) to extend the residence time of the reactants in the catalytic zone; in the late stage of the reaction, the channel width can be restored to a larger value to accelerate the discharge of the products. Example 4

[0043] The microchannel reactor used for the detection of drug-resistant Brucella malta employs a parallel processing strategy, integrating multiple independent self-assembled nanocatalytic microchannel reactor reaction units on a single chip. These units, comprising injection, reaction, and detection zones, are capable of processing different samples simultaneously. For example, the microfluidic chip measures 50 mm × 30 mm × 10 mm and houses 12 parallel reaction units, each with a volume of 10 μL. The microchannel reactor utilizes a multilayered design, comprising a reaction layer, a heating layer, a cooling layer, and a sensing layer. The reaction layer comprises microchannels and a nanocatalytic structure; the heating layer contains microheating elements capable of raising the temperature by 10°C in 0.1 seconds; the cooling layer comprises microPeltier elements capable of lowering the temperature by 10°C in 0.2 seconds; and the sensing layer includes various microsensors for real-time monitoring of reaction parameters. The self-assembled nanocatalytic microchannel reactor enables high-throughput sample processing, increasing processing capacity by 16 times compared to traditional methods.

[0044] The microfluidic chip, integrated with a self-assembled nanocatalytic microchannel reactor, adopts a modular design, making the system highly flexible and scalable. The entire system is divided into two parts: a basic platform and functional modules. The basic platform includes a power supply system, a control system, and a communication system to provide support for each functional module. The functional modules include reaction modules, separation modules, detection modules, and storage modules, which can be combined and replaced as needed. The modules are connected via standard interfaces and use a plug-and-play method, eliminating the need for complex installation and debugging. For example, when different types of catalytic reactions are required, only the reaction module needs to be replaced, while the other modules remain unchanged. When different types of products need to be detected, only the detection module needs to be replaced, while the other modules remain unchanged. The modular design greatly improves the system's adaptability, enabling it to handle a variety of complex reaction and separation tasks. Example 5

[0045] The microchannel reactor used for drug-resistant Brucella malta detection can also integrate intelligent control software, for example, using a modular design consisting of a data acquisition module, a parameter control module, a data analysis module, and a user interface module. The data acquisition module is responsible for collecting reaction parameter data from various sensors, with a sampling frequency of 10 Hz. The parameter control module is responsible for adjusting the microchannel reactor's operating parameters, such as flow rate, temperature, and pressure. The data analysis module uses machine learning algorithms to perform real-time analysis of collected data, identifying reaction trends and anomalies. The user interface module provides a user-friendly interface that displays reaction status and analysis results. The intelligent control software can precisely manage the flow rate and temperature of multiple reaction units, with a flow rate control accuracy of ±0.01 μL / min and a temperature control accuracy of ±0.1°C. Through the coordinated management of the intelligent control software, sample processing time is reduced from several hours using traditional methods to less than 30 minutes, significantly improving work efficiency.

[0046] The intelligent control algorithm, based on machine learning technology, comprises four modules: data preprocessing, feature extraction, model training, and decision execution. The data preprocessing module filters, normalizes, and detects outliers in the raw data collected by the sensors. The feature extraction module extracts time-domain, frequency-domain, and statistical features from the preprocessed data. The model training module uses a deep neural network algorithm to establish a mapping relationship between reaction parameters and product quality based on historical data. The decision execution module automatically adjusts parameters such as microchannel shape, flow rate, and temperature based on the model's predictions. The intelligent control algorithm's learning capability enables the system to continuously optimize operating parameters, improving reaction efficiency and product quality. During the system's initial operation, the reaction conversion rate was 85%. After 100 reaction cycles of learning, the reaction conversion rate increased to 95%, and product selectivity increased from 80% to 92%. Example 6

[0047] In this embodiment, the microchannel reactor used for the detection of drug-resistant Brucella malta is specially optimized for bacterial resistance detection. For example, the self-assembled nanocatalytic microchannel reactor is integrated into a microfluidic chip with a size of 40mm×25mm×5mm. It is equipped with four functional areas: sample pretreatment area, bacterial culture area, antibiotic gradient area and detection area. The sample pretreatment area contains a microfiltration device and a lysis device, which can separate bacteria from complex samples and lyse them; the bacterial culture area contains a microculture chamber and a temperature control unit, which can quickly culture bacteria under optimal conditions; the antibiotic gradient area contains a microgradient generator, which can produce 8 different concentrations of antibiotic gradients; the detection area contains a fluorescence detector and an electrochemical detector, which can monitor the growth of bacteria in real time. The self-assembled nanocatalytic microchannel reactor shortens the bacterial resistance detection cycle from 24-48 hours of traditional methods to 2-12 hours, improving the detection efficiency by more than 50%.

[0048] The detection area includes a gene detection unit, which is a multimodal sensor array, including a micro mass spectrometer sensor, a fluorescence sensor, an electrochemical sensor, a real-time PCR and / or CRISPR-Cas. For example, an online micro mass spectrometer is integrated at the outlet of the self-assembled nanocatalytic microchannel reactor to monitor sample processing and reaction progress in real time. The size of the micro mass spectrometer is 100mm×80mm×50mm, with a mass range of 50-2000m / z, a resolution of 0.1m / z, and a sampling frequency of 1Hz. The micro mass spectrometer uses electrospray ionization technology and can directly analyze liquid samples without the need for additional sample processing steps. The micro mass spectrometer is connected to the control unit via a data cable, and the analysis results are transmitted to the intelligent control software in real time for processing. The online micro mass spectrometer ensures the accuracy and timeliness of the data, making the entire system more intelligent and automated, meeting the needs of high-throughput detection. It can also integrate fluorescence sensors using fiber-optic coupling technology, with an excitation wavelength of 365nm, a detection wavelength range of 400-600nm, and a sensitivity of 1nM; or integrate electrochemical sensors using microelectrode array technology, containing 16 independent working electrodes, with a potential range of -2V to +2V and a current measurement range of 1nA to 1mA.

[0049] These sensors are distributed across the microchannel reactor, forming a sensor network that comprehensively monitors reaction progress and product quality. The multimodal sensor array provides a wealth of reaction information, enabling the system to more comprehensively understand the reaction state and make more accurate control decisions.

[0050] To address the issues of lengthy sample processing times and complex operations, the system optimized software performance and adopted advanced microfluidic design. Software optimization encompasses both algorithm and interface optimization. Algorithm optimization utilizes parallel computing technology, increasing data processing speed by three times; interface optimization employs ergonomic principles to simplify operational procedures and reduce the number of user steps. Microfluidic design optimization encompasses both channel structure and material optimization. Channel structure optimization utilizes a three-dimensional structure to increase channel density per unit area and enhance processing capacity; material optimization utilizes novel biocompatible materials to reduce sample adsorption and cross-contamination. Through these optimization measures, the platform achieves more efficient drug resistance testing, reducing detection time by 60% and operational steps by 70% compared to traditional methods.

[0051] This has resulted in a complete system for detecting antibiotic resistance in Brucella malta. This system comprises a bottom hardware layer, primarily composed of a microfluidic chip, including a self-assembled nanocatalytic microchannel reactor and sensors; a middle control layer; and a top software layer. Each layer communicates via standard interfaces, achieving seamless integration. This multi-layered technical architecture makes the system highly flexible and scalable, adapting to diverse detection needs.

[0052] The above are merely specific embodiments of the present invention, but the scope of protection of the present invention is not limited thereto. Any modifications or substitutions that can be easily conceived by a person skilled in the art within the technical scope disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be based on the scope of protection of the claims.

Claims

1. A microchannel reactor for detecting drug-resistant Brucella malta, characterized in that: The microchannel reactor comprises a microchannel body, a circulating microchannel structure, a microsensor system, a control unit and a gene detection unit; The microchannel body is made of a polymer material, including one or more of polydimethylsiloxane (PDMS), polymethyl methacrylate (PMMA), polytetrafluoroethylene (PTFE), polycarbonate (PC) or cyclic olefin copolymer (COC); The microchannel body is provided with multiple microchannels, each with a diameter of 50 to 500 μm, and an inlet and an outlet are provided at both ends of the microchannel body. The surface of the microchannel has a hydrophilic and hydrophobic pattern, which is achieved by a combination of photolithography and / or surface modification to guide the fluid to flow along a preset path. The microchannels are connected to a spiral-shaped circulation microchannel structure, and a nanocatalytic structure layer is constructed on the inner wall surface of the circulation microchannel structure, which is prepared by layer-by-layer self-assembly technology; The micro sensor system is integrated at a key position of the microchannel reactor, and includes one or more of a temperature sensor, a pressure sensor, a pH sensor and / or an optical sensor; The control unit adjusts operating parameters and / or optimizes reaction conditions based on data fed back by the microsensor system; The gene detection unit is a multimodal sensor array, including a micro mass spectrometry sensor, a fluorescence sensor, an electrochemical sensor, real-time PCR and / or CRISPR-Cas.

2. The microchannel reactor according to claim 1, wherein The surface of the microchannel is scanned in three dimensions controlled by a computer using a femtosecond laser system to create a complex micro-nano structure, which is combined with chemical modification to form a hydrophilic-hydrophobic pattern to control the flow path of the fluid; the circulation microchannel structure includes one or more micro-mixing units.

3. The microchannel reactor according to claim 1, wherein The microchannel contains a multi-layer filtration structure composed of membranes with different functions. The membranes with different functions are formed of a membrane substrate formed by a blend of polysulfone (PSF) and polyethersulfone (PES). The mass ratio of polysulfone to polyethersulfone is 7:

3. The nano-enhancer is selected from carbon nanotubes and / or silica nanoparticles, which are prepared by a phase inversion method. The separation efficiency and selectivity are improved through different pore sizes and chemical modifications.

4. The microchannel reactor according to claim 1, wherein The microchannel is formed of a deformable microchannel structure using piezoelectric material and shape memory alloy, so that the width of the microchannel can be continuously changed within the range of 50-500 μm; The piezoelectric material is polyvinylidene fluoride (PVDF) to form a piezoelectric actuator, and the shape memory alloy is nickel-titanium alloy, which is distributed at key positions of the microchannel; When voltage is applied to the piezoelectric actuator, the piezoelectric material deforms, pushing the microchannel wall to deform; when the shape memory alloy is heated, the alloy restores the preset shape, pulling the microchannel wall to restore.

5. The microchannel reactor according to claim 1, wherein One or more of the microchannel reactors are integrated on a chip, comprising an injection area, a reaction area and a detection area; the microchannel reactor adopts a multi-layer structure design, including a reaction layer, a heating layer, a cooling layer and a sensing layer, the reaction layer includes the microchannel body and the circulating microchannel structure, the heating layer includes a micro heating element, the cooling layer includes a micro Peltier element, and the sensing layer includes one or more micro sensors.

6. The microchannel reactor according to claim 5, wherein The chip adopts a modular design, including a basic platform and functional modules; the basic platform includes a power system, a control system and a communication system, and the functional modules include a reaction module, a separation module, a detection module and / or a storage module; each module adopts a plug-and-play method and can be replaced according to separation or detection needs.

7. The microchannel reactor according to claim 1, wherein The microchannel reactor integrates intelligent control software with a modular design, including a data acquisition module, a parameter control module, a data analysis module and / or a user interface module; the data acquisition module obtains reaction data from each sensor in the microsensor system; the parameter control module provides feedback to adjust the operating parameters of the microchannel reactor, the data analysis module performs real-time analysis of the collected data, and the user interface module provides a user-friendly operation interface to display the reaction status and analysis results.

8. The microchannel reactor according to claim 1, wherein The microchannel reactor is used for bacterial resistance detection of Brucella malta to form a bacterial resistance detection system; the bacterial resistance detection system includes a hardware layer, the hardware layer includes a microfluidic chip, which is integrated by the microchannel reactor, including one or more functional areas in the sample pretreatment area, bacterial culture area, antibiotic gradient area and detection area; the detection area includes a gene detection unit, which is a multimodal sensor array, including a micro mass spectrometry sensor, a fluorescence sensor, an electrochemical sensor, real-time PCR and / or CRISPR-Cas, distributed at different positions of the microchannel reactor.

9. The microchannel reactor according to claim 8, wherein The bacterial resistance detection system also includes a control layer and a software layer. The control layer includes a microprocessor and / or a drive circuit. The software layer adopts a microfluidic design, including channel structure optimization and / or material optimization. The channel structure optimization adopts a three-dimensional structure, and the material optimization adopts a biocompatible material.

10. The microchannel reactor according to claim 1, wherein The reactants enter the microchannel through the feed port and flow along a preset path under the guidance of the hydrophilic and hydrophobic patterned surface of the microchannel; after entering the circulation microchannel structure, the reactants undergo a catalytic reaction under the action of the nanocatalytic structure layer, the reaction parameters are monitored in real time by the microsensor system, and the reaction conditions are adjusted by the control unit, and the reaction products finally flow out from the discharge port and are collected.

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