Bacteria detection micro-fluidic chip system in gas driving mode

Abstract

The utility model discloses a gas-driven micro-fluidic chip system for detecting bacteria, which comprises a micro-fluidic chip, and the micro-fluidic chip comprises three layers of structures, namely a gas flowing layer, a thin film and a liquid flowing layer from top to bottom; the vacuum system comprises a negative pressure pump and a plurality of guide pipes; the pressure system comprises two gas conveying pumps and a plurality of guide pipes, and the gas conveying pumps comprise an airflow layer gas conveying pump and a liquid flow layer gas conveying pump; a solenoid valve; the intelligent control system comprises a controller and a plurality of wires; the plurality of leads are respectively connected with the electromagnetic valve, the negative pressure pump and the air delivery pump, and the electromagnetic valve, the negative pressure pump and the air delivery pump are controlled to work by inputting a programming program into the controller. The system is low in cost, and has the advantages of high detection efficiency, multiple bacteria joint detection, intelligence and automation.

Description

Technical Field

[0001] The utility model relates to the technical field of bacterial detection systems, in particular to a bacterial detection microfluidic chip system driven by gas. Background Art

[0002] Bacterial bloodstream infection is a common infectious disease in clinical practice. When pathogenic bacteria invade the blood, they will rapidly multiply, spread throughout the body through blood circulation, release toxins, and can cause systemic infection. The disease develops rapidly and can cause septic shock or multiple organ damage in severe cases, seriously threatening the health of patients. Therefore, rapid diagnosis of pathogenic bacteria is crucial for reducing the mortality of bloodstream infection and guiding clinical treatment.

[0003] A Chinese patent with the publication number CN118272549A discloses a detection method for bacteria in exhaled breath and the equipment used therefor. The method includes the following steps: S1, enriching microorganisms in exhaled breath; S2, eluting microorganisms; S3, extracting microbial nucleic acids from the eluate by electrolysis; S4, RPA isothermal amplification; S5, detecting the result by lateral flow analysis experiment. However, this method has problems such as complex operation steps, few detected bacterial species, and low detection sensitivity.

[0004] A Chinese patent with the publication number CN114480096A discloses a pressing type microfluidic chip, a microfluidic device and a bacterial detection method. This method changes the pressure in the corresponding control cavity by pressing the operation cavity with fingers and acts on a preset position of the detection liquid path to control the flow of the liquid in the detection liquid path, so as to complete the detection of bacteria. However, this detection method integrates the processes of bacterial identification, capture, lysis, purification, amplification, signal output, etc. on a single chip, but the entire operation process still requires manual operation, with low automation and consuming a large amount of time and manpower.

[0005] In summary, the existing bacterial detection technologies for bloodstream infection on the market have problems such as complex operation steps and low automation. Therefore, it is of important practical value to research and develop a rapid detection bacterial detection system with convenient operation, low cost, high efficiency and intelligence. Summary of the Utility Model

[0006] The purpose of the utility model is to provide a bacterial detection microfluidic chip system driven by gas, which has the advantages of low cost, high detection efficiency, multiple bacterial joint detections and intelligent automation.

[0007] A bacterial detection microfluidic chip system driven by gas includes:

[0008] A microfluidic chip, the microfluidic chip includes a three-layer structure, which are a gas flow layer, a thin film, and a liquid flow layer from top to bottom. The liquid flow layer is provided with a plurality of liquid channels, and the gas flow layer is provided with a plurality of gas channels. Part of the liquid channels and the gas channels and the thin film form a pneumatic valve. An air inlet hole is opened on the gas flow layer. The gas channels cooperate with the thin film to control the opening and closing of the pneumatic valve, and further control the liquid flow. The liquid flow layer is sequentially provided with an enrichment area, an amplification area, and a cutting fluorescence area at preset intervals;

[0009] A vacuum system, the vacuum system includes a negative pressure pump and a plurality of conduits;

[0010] A pressure system, the pressure system includes two gas delivery pumps and a plurality of conduits. The gas delivery pumps include an air flow layer gas delivery pump and a liquid flow layer gas delivery pump;

[0011] A solenoid valve, the number of solenoid valves is multiple. The solenoid valve is provided with an air inlet hole, an air outlet hole, and an adjustment hole. The air inlet holes of the solenoid valve are respectively connected to the negative pressure pump or the gas delivery pump through conduits. The air outlet hole is connected to the working area through a conduit. The adjustment hole is used in cooperation with the air outlet hole and the air inlet hole to control the conduction of gas delivery and air extraction;

[0012] An intelligent control system, the intelligent control system includes a controller and a plurality of wires; the plurality of wires are respectively connected to the solenoid valve, the negative pressure pump, and the gas delivery pump. By inputting a programming program into the controller, the solenoid valve, the negative pressure pump, and the gas delivery pump are controlled to work.

[0013] Compared with the prior art, the bacterial detection microfluidic chip system with a gas driving method disclosed by the present invention has the following beneficial effects:

[0014] 1. Driving innovation. The present invention uses a microfluidic chip with a gas driving method for bacterial detection. Compared with the motor used in the centrifugal driving method, the air pump used in this method has the advantages of simple structure, low cost, and convenient operation. In addition, the air pump operates silently, has a low maintenance cost, and has a longer service life and higher reliability, and is suitable for application environments that require long-term operation.

[0015] 2. Functional innovation. The microfluidic chip designed by the present invention covers the entire process of bacterial detection, including bacterial enrichment, nucleic acid amplification, nucleic acid cutting, and color reaction, realizing integrated detection. The integrated design has the following advantages:

[0016] 1) High detection efficiency. The microfluidic technology can detect bacteria at extremely low concentrations, without the need for pretreatment and cultivation of samples, greatly saving the time for bacterial cultivation and realizing rapid detection of bacteria;

[0017] 2) Multiple bacteria can be jointly detected. There are multiple cutting and color development areas on the microfluidic chip, enabling the joint detection of multiple bacteria; there are multiple channels in the cutting and color development areas of the microfluidic chip to respectively detect the housekeeping, drug resistance, and virulence genes of the same kind of bacteria, greatly solving the problem of few joint bacteria detections and incomplete results in the current market.

[0018] 3) Intelligent automation. After programming the corresponding program, the intelligent control system can automatically control the operation of the entire microfluidic chip system, greatly improving the processing speed and efficiency of bacteria, saving time and human resources, and reducing the risk of human misoperation. In addition, it can be programmed and adjusted according to different needs, with high flexibility and adaptability.

[0019] Therefore, in this utility model, the pneumatic valve is controlled to open and close through the pressure system and the vacuum system. The pressure system applies air pressure to the liquid flow layer to drive the sample liquid to flow. The liquid flows through the enrichment area, the amplification area, and the cutting and fluorescence area through the pneumatic valve, and finally the bacteria concentration of the sample liquid is measured through fluorescence detection. The bacteria detection microfluidic chip system with a gas-driven method of this utility model has a simple structure and low cost, and has the advantages of high detection efficiency, multiple joint bacteria detections, and intelligent automation.

[0020] As a further improvement of the above technical solution, the enrichment area and the amplification area are fan-shaped areas close to the center of the microfluidic chip. The enrichment area and the amplification area are symmetrically distributed about the central axis of the microfluidic chip, and the enrichment area is connected to the amplification area through pneumatic valve 1.

[0021] As a further improvement of the above technical solution, the amplification area is connected to the annular liquid storage chamber through pneumatic valve 2, and the annular liquid storage chamber is connected to the cutting and fluorescence chamber through multiple pneumatic valves; the multiple pneumatic valves are evenly distributed around the central axis of the microfluidic chip; there are multiple cutting and fluorescence chambers, and each cutting and fluorescence chamber includes three detection areas to respectively detect the housekeeping, drug resistance, and virulence genes of the same kind of bacteria.

[0022] As a further improvement of the above technical solution, the enrichment area uses the functionalized nanomagnetic bead enrichment technology; the amplification area uses the RPA amplification technology; the cutting and fluorescence area uses the CRISPR-AIEgens cutting and fluorescence technology.

[0023] As a further improvement of the above technical solution, the gas flow layer and the film are provided with through holes at the corresponding positions of the enrichment area of the liquid flow layer, and the sample liquid is injected into the enrichment area through the through holes.

[0024] As a further improvement of the above technical solution, the through hole is simultaneously an air inlet hole of the liquid flow layer. The through hole is connected to the air outlet hole of the first electromagnetic valve through a conduit. The air inlet hole of the first electromagnetic valve is connected to the air delivery pump of the liquid flow layer through a conduit. By controlling the opening and closing of the first electromagnetic valve, an intermittent pressure is applied to the sample liquid, driving the sample liquid to sequentially pass through each pneumatic valve according to the working requirements, and flowing through the enrichment area, the amplification area, and the cutting fluorescence area.

[0025] As a further improvement of the above technical solution, the microfluidic chip is provided with a plurality of the pneumatic valves. The air inlet hole of each pneumatic valve is respectively connected to two electromagnetic valves through conduits. The first pneumatic valve is connected to the air outlet holes of the second electromagnetic valve and the third electromagnetic valve. The air inlet hole of the second electromagnetic valve is connected to the air delivery pump of the air flow layer through a conduit. By controlling the opening and closing of the second electromagnetic valve, an intermittent pressure is applied to the thin film, causing the first pneumatic valve to close. The air inlet hole of the third electromagnetic valve is connected to the negative pressure pump through a conduit. By controlling the opening and closing of the third electromagnetic valve, the thin film is restored, causing the first pneumatic valve to open.

[0026] As a further improvement of the above technical solution, the thin film has good elasticity and can undergo good elastic deformation under pressure.

[0027] As a further improvement of the above technical solution, the microfluidic chip system further includes a fluorescence detection device. The fluorescence detection device is located directly below the microfluidic chip. The fluorescence detection device is connected to the controller through a wire, and the detection result is presented through the display screen of the controller. Description of the Drawings

[0028] The above and / or additional aspects and advantages of the present utility model will become apparent and easy to understand from the description of the embodiments in conjunction with the following drawings, where:

[0029] Figure 1 is the working principle diagram of the bacterial detection microfluidic chip system with a gas driving mode of the present utility model;

[0030] Figure 2 is the working flow chart of the bacterial detection microfluidic chip system with a gas driving mode of the present utility model;

[0031] Figure 3 is the working principle diagram of the pneumatic valve in the bacterial detection microfluidic chip system with a gas driving mode of the present utility model;

[0032] Figure 4 is the structural schematic diagram of the bacterial detection microfluidic chip system with a gas driving mode of the present utility model;

[0033] Figure 5Schematic diagram of the microfluidic chip structure in the bacterial detection microfluidic chip system with gas drive mode of the present utility model;

[0034] Figure 6 Schematic diagram of the air flow layer structure in the bacterial detection microfluidic chip system with gas drive mode of the present utility model;

[0035] Figure 7 Schematic diagram of the liquid flow layer structure in the bacterial detection microfluidic chip system with gas drive mode of the present utility model.

[0036] Figure 8 Schematic diagram of the through-hole wiring in the bacterial detection microfluidic chip system with gas drive mode of the present utility model.

[0037] Figure 9 Schematic diagram of the pneumatic valve 1 wiring in the bacterial detection microfluidic chip system with gas drive mode of the present utility model.

[0038] Figure 10 Bacterial detection result of the display screen in the bacterial detection microfluidic chip system with gas drive mode of the present utility model. Detailed implementation manners

[0039] To make the objectives, features and advantages of the present utility model more obvious and understandable, the following detailed description of the specific implementation manners of the present utility model is given with reference to the accompanying drawings. Several embodiments of the present utility model are shown in the drawings. However, the present utility model can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, these embodiments are provided to make the disclosure of the present utility model more thorough and comprehensive.

[0040] It should be noted that when an element is referred to as being "fixedly provided on" another element, it can be directly on the other element or there may also be an intermediate element. When an element is considered to be "connected" to another element, it can be directly connected to the other element or there may be an intermediate element at the same time. The terms "vertical", "horizontal", "left", "right", "up", "down" and similar expressions used herein are only for the purpose of illustration and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and thus cannot be construed as a limitation of the present utility model.

[0041] In the present utility model, unless otherwise clearly defined and limited, terms such as "installation", "connection", "linkage", "fixation", etc. shall be understood in a broad sense. For example, it may be a fixed connection, a detachable connection, or an integral connection; it may be a mechanical connection or an electrical connection; it may be a direct connection or an indirect connection through an intermediate medium, and it may be the communication inside two components. For those of ordinary skill in the art, the specific meanings of the above terms in the present utility model can be understood according to specific circumstances. The term "and / or" used herein includes any and all combinations of one or more related listed items.

[0042] As Figures 1 to 9 shown, the present utility model provides a bacterial detection microfluidic chip system with a gas driving mode, including:

[0043] A microfluidic chip 1, the microfluidic chip includes a three-layer structure, which are, from top to bottom, a gas flow layer 11, a thin film 12, and a liquid flow layer 13. The liquid flow layer is provided with a plurality of liquid channels 131, and the gas flow layer is provided with a plurality of gas channels 111; some of the liquid channels 131 and gas channels 111 and the thin film 12 form a pneumatic valve 14; an air inlet hole is opened on the gas flow layer 11; the gas channels 111 cooperate with the thin film 12 to control the opening and closing of the pneumatic valve 14, and further control the liquid flow. The liquid flow layer 13 is sequentially provided with an enrichment area 132, an amplification area 133, and a cutting fluorescence area 134 at certain intervals;

[0044] A vacuum system 2, the vacuum system includes a negative pressure pump 21 and a plurality of conduits;

[0045] A pressure system 3, the pressure system includes two gas delivery pumps 31 and a plurality of conduits, and the gas delivery pumps include a gas flow layer gas delivery pump 311 and a liquid flow layer gas delivery pump 312;

[0046] Electromagnetic valves 4, the number of electromagnetic valves 4 is multiple, the electromagnetic valves are provided with air inlet holes, air outlet holes, and adjustment holes, the air inlet holes of each electromagnetic valve are respectively connected to the negative pressure pump 21 or the gas delivery pump 31 through conduits, the air outlet holes are connected to the working area through conduits, and the adjustment holes cooperate with the air outlet holes and air inlet holes to control the on-off of gas delivery and air extraction;

[0047] An intelligent control system 5, the intelligent control system includes a controller 51 and a plurality of wires; the plurality of wires are respectively connected to the electromagnetic valve 3, the negative pressure pump 211, and the gas delivery pump 311, and by inputting a programming program into the controller 51, the electromagnetic valve 3, the negative pressure pump 21, and the gas delivery pump 31 are controlled to work.

[0048] The utility model relates to a bacterial detection microfluidic chip system with a gas driving mode. The opening and closing of a pneumatic valve 14 are controlled by a pressure system 2 and a vacuum system 3. An air pressure is applied to a liquid flow layer 13 by the pressure system 2 to drive the flow of a sample liquid. The liquid flows through an enrichment area 132, an amplification area 133, and a cleavage fluorescence area 134 through the pneumatic valve 14, and finally the bacterial concentration of the sample liquid is measured by fluorescence detection. The bacterial detection microfluidic chip system with the gas driving mode of the utility model has a simple structure and low cost, and has the advantages of high detection efficiency, multiple joint detections of bacteria, and intelligent automation.

[0049] In some specific embodiments, a gas channel 111 is perpendicular to a liquid channel 131, and thus forms a pneumatic valve 14 with a thin film 12.

[0050] In some specific embodiments, the number of pneumatic valves 14 is seven, namely a first pneumatic valve 141, a second pneumatic valve 142, and five pneumatic valves 143; the number of electromagnetic valves 14 is seven.

[0051] In some specific embodiments, the enrichment area 132 and the amplification area 133 are fan-shaped areas close to the center of the microfluidic chip 1. The enrichment area 132 and the amplification area 133 are symmetrically distributed about the central axis of the microfluidic chip 1, and the enrichment area 132 is connected to the amplification area 133 through the first pneumatic valve 141.

[0052] In some specific embodiments, the amplification area 133 is connected to an annular liquid storage chamber 1311 through the second pneumatic valve 142. The annular liquid storage chamber 1311 is connected to a cleavage fluorescence chamber 134 through five pneumatic valves 143. The five pneumatic valves 143 are evenly distributed around the central axis of the microfluidic chip 1; there are five cleavage fluorescence chambers 134, and each cleavage fluorescence chamber 134 includes three detection areas for respectively detecting the housekeeping, drug resistance, and virulence genes of the same kind of bacteria.

[0053] In some specific embodiments, the enrichment area 132 uses the technology of enriching concanavalin A-functionalized magnetic beads; the amplification area 133 uses the multiplex RPA amplification technology; the cleavage fluorescence area 134 uses the CRISPR / Cas12a-AIEgens fluorescence technology.

[0054] In some specific embodiments, through holes 15 are provided at corresponding positions of the gas flow layer 11 and the thin film 12 in the enrichment area 132 of the liquid flow layer 13, and the sample liquid is injected into the enrichment area 132 through the through holes 15.

[0055] In some specific embodiments, the through-hole 15 serves as the air inlet hole of the liquid flow layer at the same time. The through-hole 15 is connected to the air outlet hole 411 of the first electromagnetic valve 41 through a conduit. The air inlet hole 412 of the first electromagnetic valve 41 is connected to the air delivery pump 312 of the liquid flow layer through a conduit. By controlling the opening and closing of the first electromagnetic valve 41, the intermittent pressure is applied to the sample liquid, driving the sample liquid to sequentially pass through the first pneumatic valve 141, the second pneumatic valve 142 and the five pneumatic valves 143 according to the working requirements, and flowing through the enrichment area 132, the amplification area 133 and the fluorescence cutting area 134.

[0056] In some specific embodiments, the air inlet hole of the first pneumatic valve 141 is connected to the air outlet holes of the second electromagnetic valve 42 and the third electromagnetic valve 43 respectively through conduits; the air inlet hole 422 of the second electromagnetic valve is connected to the air delivery pump 311 of the air flow layer through a conduit. By controlling the opening and closing of the second electromagnetic valve 42, the intermittent pressure is applied to the thin film 12 to close the first pneumatic valve 141; the air inlet hole 432 of the third electromagnetic valve is connected to the negative pressure pump 21 through a conduit. By controlling the opening and closing of the third electromagnetic valve 43, the thin film 12 is restored and the first pneumatic valve 141 is opened.

[0057] Specifically, the second pneumatic valve 142 is connected to the fourth electromagnetic valve 44 and the fifth electromagnetic valve 45; the five pneumatic valves are connected to the sixth electromagnetic valve 46 and the seventh electromagnetic valve 47; the working principles of the second pneumatic valve and the five pneumatic valves are the same as those of the first pneumatic valve.

[0058] In some specific embodiments, the thin film 12 has good elasticity and can be elastically deformed well under pressure.

[0059] In some specific embodiments, the microfluidic chip system further includes a fluorescence detection device 6. The fluorescence detection device 6 is located directly below the microfluidic chip 1. The fluorescence detection device 6 is connected to the controller 51 through a wire, and the detection result is presented on the display screen of the controller 51.

[0060] The working steps of the bacterial detection microfluidic chip system with a gas driving mode of the present utility model are as follows:

[0061] S1. Add samples to the enrichment area through the through-hole.

[0062] S2. Control the pressure system to supply air to the liquid flow layer so that the bacteria in the sample liquid are fully enriched.

[0063] S3. Control the first pneumatic valve to open, control the pressure system to supply air to the liquid flow layer, and the sample liquid enters the amplification area so that the bacteria in the sample liquid are fully amplified.

[0064] S4. Control the second pneumatic valve to open, control the pressure system to supply gas to the liquid flow layer, and the sample liquid enters the annular liquid flow channel. Control five pneumatic valves to open, control the pressure system to supply gas to the liquid flow layer, and the sample liquid is evenly divided into five parts and enters five cutting fluorescence regions respectively. Each cutting fluorescence region is further divided into three detection regions to detect the housekeeping, drug resistance, and virulence genes of bacteria respectively. Then, the bacteria concentration is measured by a fluorescence detection device, and the detection result is presented on the display screen. An exemplary detection result is as Figure 10 shown.

[0065] The present utility model adopts a variety of technologies in the liquid flow layer, including functionalized nanomagnetic bead enrichment technology in the enrichment region, multiplex RPA technology in the amplification region, and CRISPR / Cas12a-AIE cutting fluorescence technology in the cutting fluorescence region.

[0066] The functionalized nanomagnetic bead enrichment technology uses concanavalin A lectin, which greatly improves the bacteria capture rate. This technology first conjugates streptavidin on the surface of nanomagnetic beads, and then conjugates biotinylated concanavalin A on the streptavidin-coated nanomagnetic beads to form a concanavalin A-nanomagnetic bead complex. Finally, the sugar recognition domain on concanavalin A specifically recognizes the mannose-type glycosyl groups on the surface of bacteria, realizing the efficient enrichment of bacteria. This technology can complete the enrichment of bacteria within 15 minutes, and the capture rate reaches 97.6%.

[0067] The multiplex RPA nucleic acid amplification technology can amplify the target DNA at room temperature. A pair of specific primers for the target DNA are designed for the characteristic genes, virulence genes, and drug resistance genes of different types of bacteria respectively. The reaction lasts for 15 - 20 minutes under isothermal conditions (37 - 42°C), and the in vitro nucleic acid amplification can reach 109 - 1010 times, and its specificity can reach 99.3%.

[0068] The CRISPR-AIEgens cleavage fluorescence technology features high specificity, high accuracy, and convenience. In the process of technology implementation, CRISPR / Cas12a cleavage and the release of the dsDNA@AIEgens fluorescent probe generate fluorescence signals. The reaction process is as follows: the crRNA in the CRISPR / Cas12a system specifically recognizes the target DNA, cis-cleaves the double-stranded DNA, and then induces the activation of the Cas protease. The Cas protein trans-cleaves the single-stranded DNA (ssDNA) in the AIEgens fluorescent probe, thus separating the dsDNA quenching group of the AIEgens fluorescent probe, and the dsDNA@AIEgens emits a strong fluorescence signal. The CRISPR / Cas12a cleavage and the release of the dsDNA@AIEgens fluorescent probe work almost synchronously, and fluorescence signals can be generated within 3 minutes, with high detection efficiency; the cleavage technology selects genes that are conserved and specific among pathogens, greatly improving the specificity; the fluorescence color development technology realizes the amplification of the ratio of the quenching group to the luminescent group as 1:N, greatly improving the accuracy.

[0069] Compared with the prior art, the bacterial detection microfluidic chip system with a gas-driven mode disclosed by the present utility model has the following beneficial effects:

[0070] 1. Driving innovation. The present utility model uses a microfluidic chip with a gas-driven mode for bacterial detection. Compared with the motor used in the centrifugal drive mode, the air pump used in this mode has the advantages of simple structure, low cost, and convenient operation. In addition, the air pump operates quietly, has low maintenance costs, and has a longer service life and higher reliability, making it suitable for application environments that require long-term operation.

[0071] 2. Functional innovation. The microfluidic chip designed by the present utility model covers the entire process of bacterial detection, including bacterial enrichment, nucleic acid amplification, nucleic acid cleavage, and color reaction, realizing integrated detection. This integrated design has the following advantages:

[0072] 1) High detection efficiency. The microfluidic technology can detect bacteria at extremely low concentrations without pre-treating and culturing the samples, greatly saving the time for bacterial culture and realizing the rapid detection of bacteria;

[0073] 2) Multiple bacteria can be jointly detected. Multiple cleavage and color development regions are provided on the microfluidic chip, enabling the joint detection of multiple bacteria; multiple channels are provided in the cleavage and color development regions of the microfluidic chip to respectively detect the housekeeping, drug resistance, and virulence genes of the same bacteria, greatly solving the problem of few joint detections of bacteria and incomplete results on the current market;

[0074] 3) Intelligent automation. After programming the corresponding program, the intelligent control system can automatically control the operation of the entire microfluidic chip system, greatly improving the processing speed and efficiency of bacteria, saving time and human resources, and reducing the risk of human error. In addition, it can be programmed and adjusted according to different needs, with high flexibility and adaptability.

[0075] Therefore, the present utility model controls the opening and closing of the pneumatic valve through the pressure system and the vacuum system, applies air pressure in the liquid flow layer through the pressure system to drive the liquid flow of the sample liquid, the liquid flows through the enrichment area, the amplification area, and the cutting fluorescence area through the pneumatic valve, and finally measures the bacterial concentration of the sample liquid through fluorescence detection. The bacterial detection microfluidic chip system with a gas-driven mode of the present utility model has a simple structure, low cost, and has the advantages of high detection efficiency, multiple bacterial joint detections, and intelligent automation.

[0076] In the description of this specification, the description with reference to terms such as "one embodiment", "some embodiments", "example", "specific example", or "some examples" means that the specific features, structures, materials, or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present utility model. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described can be combined in any one or more embodiments or examples in a suitable manner.

[0077] The above-described embodiments only represent several implementation manners of the present utility model, and their descriptions are relatively specific and detailed, but should not be construed as a limitation on the scope of the patent of the present utility model. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present utility model, several modifications and improvements can still be made, and these all belong to the protection scope of the present utility model. Therefore, the protection scope of the patent of the present utility model should be subject to the appended claims.

Claims

1. A gas-driven bacteria detection microfluidic chip system, comprising: A microfluidic chip, wherein the microfluidic chip comprises a three-layer structure, which comprises a gas flow layer, a thin film, and a liquid flow layer from top to bottom. The liquid flow layer is provided with a plurality of liquid channels, and the gas flow layer is provided with a plurality of gas channels. Part of the liquid channels, the gas channels, and the thin film form a pneumatic valve. An air inlet is provided on the gas flow layer. The gas channel cooperates with the thin film to control the opening and closing of the pneumatic valve, thereby controlling the flow of liquid. The liquid flow layer is provided with an enrichment area, an amplification area, and a cutting fluorescence area in sequence at preset intervals. A vacuum system, the vacuum system comprising a negative pressure pump and a plurality of catheters; A pressure system, the pressure system comprising two air pumps and a plurality of conduits, the air pumps comprising an air flow layer air pump and a liquid flow layer air pump; Solenoid valves, the number of which is multiple, the solenoid valves are provided with an air inlet, an air outlet and an adjusting hole, the air inlet of the solenoid valve is connected to the negative pressure pump or the air delivery pump through a conduit, the air outlet is connected to the working area through a conduit, and the adjusting hole cooperates with the air outlet and the air inlet to control the conduction of air delivery and air extraction; An intelligent control system, the intelligent control system includes a controller and multiple wires; the multiple wires are respectively connected to the solenoid valve, the negative pressure pump, and the air pump, and the solenoid valve, the negative pressure pump, and the air pump are controlled to work by inputting a programming program into the controller.

2. The gas-driven bacteria detection microfluidic chip system according to claim 1, characterized in that: The enrichment area and the amplification area are fan-shaped areas close to the center of the microfluidic chip. The enrichment area and the amplification area are symmetrically distributed about the central axis of the microfluidic chip. The enrichment area is connected to the amplification area through a pneumatic valve 1.

3. The gas-driven bacteria detection microfluidic chip system according to claim 2, characterized in that: The amplification zone is connected to the annular liquid storage chamber through pneumatic valve 2, and the annular liquid storage chamber is connected to the cutting fluorescence chamber through multiple pneumatic valves; the multiple pneumatic valves are evenly distributed around the central axis of the microfluidic chip; there are multiple cutting fluorescence chambers, each of which includes three detection areas, which respectively detect housekeeping, drug resistance, and virulence genes of the same bacteria.

4. The gas-driven bacteria detection microfluidic chip system according to claim 3, characterized in that: The gas flow layer and the thin film are provided with through holes at positions corresponding to the enrichment area of ​​the liquid flow layer, and the sample liquid is injected into the enrichment area through the through holes.

5. The gas-driven bacteria detection microfluidic chip system according to claim 4, characterized in that: The through hole is also the air inlet of the liquid flow layer. The through hole is connected to the air outlet of the solenoid valve 1 through a conduit. The air inlet of the solenoid valve 1 is connected to the liquid flow layer air pump through a conduit. The opening and closing of the solenoid valve 1 is controlled to achieve intermittent pressure on the sample liquid, driving the sample liquid to pass through each of the pneumatic valves in turn according to working requirements, and flow through the enrichment area, the amplification area and the cutting fluorescence area.

6. The gas-driven bacteria detection microfluidic chip system according to claim 5, characterized in that: The microfluidic chip is provided with a plurality of pneumatic valves, and the air inlet of each pneumatic valve is connected to two solenoid valves through a conduit; the pneumatic valve one is connected to the air outlet of the solenoid valve two and the air outlet of the solenoid valve three; the air inlet of the solenoid valve two is connected to the air flow layer air pump through a conduit to control the opening and closing of the solenoid valve two, thereby realizing intermittent pressure application to the film to close the pneumatic valve one; the air inlet of the solenoid valve three is connected to the negative pressure pump through a conduit to control the opening and closing of the solenoid valve three, thereby restoring the film and opening the pneumatic valve one.

7. The gas-driven bacteria detection microfluidic chip system according to claim 6, characterized in that: The microfluidic chip system also includes a fluorescence detection device, which is located directly below the microfluidic chip. The fluorescence detection device is connected to the controller via a wire, and the detection result is presented on the display screen of the controller.

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