A microfluidic sample testing device and a microfluidic sample testing system

Abstract

The application provides a microfluid sample detection device and a microfluid sample detection system, wherein the microfluid sample detection device comprises a fluid inlet, a microfluid channel, a fluid outlet, a reaction area arranged between the fluid inlet and the fluid outlet and partially overlapping with the microfluid channel, a liquid collecting area arranged downstream of the reaction area, a first pump body arranged upstream of the reaction area, a first valve body arranged downstream of the fluid inlet and controlling the on-off of the microfluid channel and the fluid inlet, a second valve body arranged downstream of the first valve body and controlling the on-off of the microfluid channel and the first pump body, and a first lock body arranged downstream of the second valve body and defining the volume of the microfluid sample together with the second valve body. The pump body provides driving force for the flow of the microfluid sample when communicating with the microfluid channel, and the working states of the first valve body and the second valve body are opposite. The microfluid sample detection device further comprises a conductive system composed of electrodes, and the conductive system comprises at least two electrodes.

Description

Technical Field

[0001] This invention relates to the field of medical devices, and more particularly to a microfluidic sample detection device and a microfluidic sample detection system. Background Technology

[0002] With the continuous development of intelligent technology, people are using more and more intelligent devices in their lives, work and study. The use of intelligent technology has improved people's quality of life and increased their learning and work efficiency.

[0003] Microfluidic chips are a scientific technology characterized by the manipulation of fluids at the micrometer scale. They possess the ability to miniaturize the basic functions of biological and chemical laboratories onto a chip of just a few square centimeters, hence the name "lab-on-a-chip." Currently, mainstream microfluidic chips primarily consist of a network of microchannels with controllable fluid flowing throughout the system to achieve various functions found in conventional chemical or biological laboratories. The fundamental characteristic and greatest advantage of microfluidic chips is the flexible combination and large-scale integration of multiple unit technologies on a tiny, controllable platform. This makes them ideally suited for the design of point-of-care testing (POCT) products. However, commonly used materials for fabricating microfluidic chips include glass and organic polymers such as polymethyl methacrylate (PMMA), polydimethylsiloxane (PDMS), polycarbonate (PC), and hydrogels. The fabrication environment is crucial, including air temperature, humidity, and particle density in the air and various media used in the preparation process. The demanding environmental requirements for chip fabrication generally necessitate a cleanroom environment. The success or failure of cleanroom technology and microfluidic chip fabrication is inextricably linked. Depending on the manufacturing method, cleanroom standards must reach Class 10,000, Class 1,000, or even Class 100. For example, the fabrication techniques used for polymer microfluidic chips mainly include hot pressing, molding, injection molding, laser ablation, and LIGA. However, the equipment used in existing technologies is expensive, and the processes are complex. These technologies not only impose strict requirements on environmental quality and incur high operating costs, but also result in low batch yields, leading to generally high product costs. This falls short of the capacity and cost requirements of point-of-care testing (POCT) products, limiting the widespread adoption of microfluidic technology in the POCT industry.

[0004] There is currently no effective solution to the above problems. Summary of the Invention

[0005] To address the aforementioned problems, the present invention provides a microfluidic sample detection device, comprising a fluid inlet, a microfluidic channel, and a fluid outlet, and:

[0006] The reaction zone is located between the fluid inlet and the fluid outlet and partially overlaps with the microfluidic channel.

[0007] The liquid collection zone is located downstream of the reaction zone, and the fluid outlet is the outlet of the liquid collection zone.

[0008] The first pump body, located upstream of the reaction zone, provides the driving force for the flow of the microfluidic sample when it is connected to the microfluidic channel;

[0009] The first valve body is located downstream of the fluid inlet and controls the opening and closing of the microfluidic channel and the fluid inlet;

[0010] The second valve body is located downstream of the first valve body and controls the opening and closing of the first pump body and the microfluidic channel. The working states of the first valve body and the second valve body are opposite.

[0011] The first locking body is located downstream of the second valve body and together with the second valve body defines the volume of the microfluidic sample.

[0012] Furthermore, the microfluidic channel includes a first region and a second region, at least the first region or the second region is a reaction zone, and the first pump can drive the microfluidic sample into the first region and the second region sequentially.

[0013] Furthermore, the microfluidic channel includes a first region, a second region, and a third region, at least the first region, the second region, or the third region is a reaction zone, and the first pump can drive the microfluidic sample to enter the first region, the second region, and the third region sequentially.

[0014] In another embodiment, the microfluidic sample detection device further includes a conductive system composed of electrodes. The conductive system comprises at least two electrodes, with sensing ends located at opposite ends of the reaction zone. Preferably, the conductive system comprises at least three electrodes, with sensing ends distributed at opposite ends of a first region and a second region. At least one of the first or second regions is a reaction zone, and the first pump can drive the microfluidic sample sequentially into the first and second regions. Further, the conductive system comprises at least four electrodes, with sensing ends distributed at opposite ends of a first, second, and third region. At least one of the first, second, or third regions is a reaction zone, and the first pump can drive the microfluidic sample sequentially into the first, second, and third regions.

[0015] In a further embodiment, the microfluidic sample detection device also includes a start electrode. The start electrode sensing end is located upstream of the second valve body and together with the first electrode sensing end downstream of it, forms a start area in the microfluidic channel. In response to the start signal generated when the microfluidic sample flows into the start area and contacts the first electrode sensing end, the first valve body closes and the second valve body opens.

[0016] Preferably, the first locking body is located in the microfluidic channel between the first electrode sensing end and the second electrode sensing end. The first electrode sensing end and the second electrode sensing end are close to the second valve body and located downstream of the second valve body, thereby ensuring that the first region and the second region are downstream of the second valve body. This ensures that the first pump body can perform its driving function while maximizing the duration of the driving force of the first pump body.

[0017] When a microfluidic sample detection device includes at least one reaction zone, in particular, the reaction zone is pre-positioned with magnetic particles that bind to antibodies against red blood cells, designed to specifically bind to red blood cells in a blood sample.

[0018] When a microfluidic sample detection device includes at least one reaction zone, in particular, the reaction zone is pre-positioned with target antibody labeled with magnetic particles, which is designed to specifically bind to the target in the sample.

[0019] When a microfluidic sample detection device includes at least one reaction zone, in particular, the reaction zone is pre-positioned with fluorescently labeled antibodies that are designed to specifically bind to targets in the sample.

[0020] When the microfluidic sample detection device includes two reaction zones, specifically, the two reaction zones are pre-positioned with a first antibody labeled with magnetic particles and a second antibody labeled with fluorescent particles. Both the first and second antibodies are designed to specifically bind to the target in the sample. After the target in the microfluidic sample binds to the first and second antibodies, a double-antibody sandwich complex is formed.

[0021] Furthermore, the microfluidic sample undergoes specific treatment before binding to the antibodies in the two reaction zones. This invention also provides a microfluidic sample detection system, including a detection device and the aforementioned microfluidic sample detection apparatus. The microfluidic sample detection apparatus is mounted within the detection device, and the detection device is electrically / signally connected to the electrodes of the microfluidic detection apparatus, thereby enabling actuators to control the opening or closing of the first and second valve bodies, as well as the compression or release of the first pump body.

[0022] Furthermore, the microfluidic sample detection device is electrically connected to the electrodes of the detection equipment via electrical contacts. The beneficial effects of this invention include: 1. The microfluidic sample detection device has a simple structure and is easy to manufacture, enabling its application in various in vitro diagnostic products using different methodologies. It can perform single-item detection, multi-item combined detection, and even combine electrochemical and photochemical detection methods into the same microfluidic sample detection device; 2. The liquid collection zone in the microfluidic sample detection device is located at the downstream end of the microfluidic channel, eliminating the problem of contaminating the detection equipment; 3. The valve and lock in the microfluidic sample detection device work together to complete sample quantification, resulting in superior accuracy and repeatability compared to traditional in vitro diagnostic electrochemical products when applied to them. Attached Figure Description

[0023] Figure 1-1 One embodiment of the microfluidic detection device of the present invention.

[0024] Figure 1-2 The second embodiment of the microfluidic detection device of the present invention.

[0025] Figure 1-3 The third embodiment of the microfluidic detection device of the present invention.

[0026] Figure 1-4 The fourth embodiment of the microfluidic detection device of the present invention.

[0027] Figure 2 The fifth embodiment of the microfluidic detection device of the present invention.

[0028] Figure 3-1 The sixth embodiment of the microfluidic detection device of the present invention.

[0029] Figure 3-2 The seventh embodiment of the microfluidic detection device of the present invention.

[0030] Figure 4-1 The eighth embodiment of the microfluidic detection device of the present invention.

[0031] Figure 4-2 The ninth embodiment of the microfluidic detection device of the present invention.

[0032] Figure 5 This invention provides a combined implementation of a microfluidic detection device.

[0033] Figure 6 One of the schematic diagrams of the microfluidic detection device module of the present invention.

[0034] Figure 7 The second schematic diagram of the microfluidic detection device module of the present invention.

[0035] Figure 8 The third schematic diagram of the microfluidic detection device module of the present invention.

[0036] Figure 9 Example 6: Results of the accuracy study of the product of the present invention.

[0037] Figure 10 Example 6A: Results of a study on the accuracy of traditional products.

[0038] Figure 11 Example 6B: Results of a study on the accuracy of conventional products. Detailed Implementation

[0039] To enable those skilled in the art to better understand the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of them. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of the present invention.

[0040] Example 1

[0041] like Figure 1-1 The diagram illustrates one embodiment of the microfluidic detection device of the present invention, comprising a fluid inlet 11, a fluid outlet 12, a microfluidic channel 13, a liquid collection zone 14, a reaction zone 131, a pump body 41, a valve body, and a lock body 21. The fluid inlet 11 is used to add the microfluidic sample to be tested. The fluid outlet 12 is in communication with the environment. The microfluidic channel 13 is located between the fluid inlet and the fluid outlet. The microfluidic sample to be tested enters from the fluid inlet and flows within the microfluidic channel. The volume of the microfluidic sample propelled forward within the microfluidic channel is the same as the volume of gas simultaneously discharged from the fluid outlet.

[0042] The liquid receiving zone 14 is located downstream of the microfluidic channel and also downstream of the reaction zone. It is used to receive the microfluidic sample after the reaction. The fluid outlet is located at the outlet of the liquid receiving zone and is in communication with the environment. The environment refers to the external environment of the microfluidic detection device of this invention, such as air or vacuum. It can be the environment set in the system of this invention or the same as the environment outside the system of this invention. The fluid outlet allows gas or liquid to flow out. In most embodiments, the fluid outlet is a gas outlet, allowing only gas to flow out and no liquid to flow out. In a few embodiments, the fluid outlet is both a gas and liquid outlet, allowing both gas and liquid to flow out.

[0043] The reaction zone 131 is located in the microfluidic channel between the fluid inlet and the fluid outlet; in other words, the reaction zone partially overlaps with the microfluidic channel. The microfluidic sample to be tested is designed to react with a fixed pre-prepared reagent in the reaction zone as it flows through the microfluidic channel.

[0044] Pump 41 is located upstream of the reaction zone and is used to passively drive the flow of the microfluidic sample in the microfluidic channel. In other words, by controlling the pressure applied to the pump, a driving force can be provided for the flow of the microfluidic sample, which drives the microfluidic sample to continue flowing downstream after entering the microfluidic channel. The pressure value applied to the pump can be preset and associated with the travel position of the microfluidic sample in the microfluidic channel after entering it. That is, when the preset pressure value applied to the pump is a certain value, the corresponding position of the microfluidic sample in the microfluidic channel is a certain position. This design can be used to confirm the successful addition of the microfluidic sample and to determine that the microfluidic sample has entered a specific region.

[0045] The system comprises two valve bodies. Valve body 31, located downstream of the fluid inlet, controls the flow between the fluid inlet and the microfluidic channel, remaining open until the microfluidic sample has fully entered the channel. Valve body 32, located downstream of valve body 31 and between the pump body and the microfluidic channel, controls the flow between the pump body and the microfluidic channel. In other words, valve body 32 controls whether the first pump body provides driving force to the microfluidic sample. Valve body 32 also helps determine the volume of the final microfluidic sample participating in the reaction. The two valve bodies operate in opposite states: valve body 31 is open when valve body 31 is closed, and vice versa. When valve body 31 is open and valve body 32 is closed, the pump body and the microfluidic channel are connected, and the pump body can passively drive the microfluidic sample to flow towards the fluid outlet.

[0046] In some implementations, the valve body consists of a plastic substrate and a flexible diaphragm. The plastic substrate includes an upper surface and a lower surface. Grooves are formed on the upper surface using methods such as laser etching. The flexible diaphragm also includes an upper surface and a lower surface. The upper surface of the plastic substrate and the lower surface of the flexible diaphragm are bonded together with an adhesive material. The upper surface of the plastic substrate and the lower surface of the flexible diaphragm are in contact and partially joined, forming a cavity between them. When the flexible diaphragm is in a relaxed state, the cavity is open, meaning the valve body is in the open state. When the flexible diaphragm is under pressure, the cavity is closed, meaning the valve body is in the closed state.

[0047] In other embodiments, the valve body is composed of a plastic substrate, double-sided adhesive, and a flexible diaphragm. The plastic substrate includes an upper surface and a lower surface, the double-sided adhesive includes a perforation, and the flexible diaphragm includes an upper surface and a lower surface. The upper surface of the plastic substrate, the double-sided adhesive, and the lower surface of the flexible diaphragm are pressed together to bond them together, forming a cavity between the upper surface of the plastic substrate and the lower surface of the flexible diaphragm. When the flexible diaphragm is in a relaxed state, the cavity is open, i.e., the valve body is in an open state. When the flexible diaphragm is under pressure, the cavity is closed, i.e., the valve body is in a closed state.

[0048] Lock body 21 is located downstream of valve body 2 and is used to limit the volume of the microfluidic sample to be tested. When the microfluidic sample to be tested enters the microfluidic channel from the fluid inlet and the front end of the microfluidic sample to be tested contacts the lock body, the flow stops. The volume of the microfluidic sample to be tested in the microfluidic channel from the fluid inlet to the lock body is the limited volume of the microfluidic sample to be tested. In a further embodiment, the lock body and valve body 2 cooperate to determine the volume of the microfluidic sample to be tested. When the microfluidic sample to be tested enters the microfluidic channel from the fluid inlet and the front end of the microfluidic sample to be tested contacts the lock body, the flow stops. After valve body 1 is closed, valve body 2 is opened, and the pump body drives the microfluidic sample to be tested from valve body 2 to the lock body to flow downstream in the microfluidic channel. This portion of the microfluidic sample flowing downstream is the limited volume of the microfluidic sample to be tested.

[0049] like Figure 6 The diagram shows the structural design of a load microfluidic detection device. To achieve the photochemical detection function, it includes a main control board, display screen, load position, magnetic module, temperature module, optical module, actuator, battery, power interface, and timer. Of course, if only a certain type of detection is required, the detection device can be simplified, for example, to only include the main control board, load position, actuator, optical module, power interface, and timer.

[0050] The power interface provides power to the testing equipment and can charge the battery. The battery allows the testing equipment to operate normally without a power source, thus achieving portability. After the testing equipment is powered on, the testing device is placed inside. When the microfluidic sample testing device is accurately placed on the load position within the testing equipment, the load position generates a load signal and transmits it to the main control board. The testing device and the testing equipment then establish a signal connection, and the main control board commands the testing equipment to start operating. In another embodiment, clicking a button on the testing device can also trigger the main control board to command the testing equipment to start operating.

[0051] The optical module includes a photodetector unit, which detects the reaction of the microfluidic sample under test. The result is then processed by the main control board and displayed on the screen. Alternatively, the optical module may also include a photoelectric conversion unit, which converts the signal into an electrical signal, which is then processed by the main control board and displayed on the screen.

[0052] On the other hand, the optical module is also used to determine whether the microfluidic sample to be tested has been successfully added to the detection device. The optical detection unit continuously detects the light reflection state at the same position in the microfluidic channel at different time points and converts it into an electrical signal by the photoelectric conversion unit. The main control board then determines that the microfluidic sample to be tested has been successfully added.

[0053] At least one reaction waiting time countdown is preset in the timer to define a fixed time period for the reaction of the microfluidic sample in the microfluidic channel. In another embodiment, a microfluidic sample addition waiting time countdown can also be set, with the time when the detection device and the detection equipment complete the signal connection as the starting point for the addition waiting time. When the timer expires, it is determined that the microfluidic sample to be tested has been successfully added. When the timer determines successful addition, it is not necessary to use the aforementioned optical detection unit to confirm successful addition. In another embodiment, a countdown can also be set for the time for the microfluidic sample to reach a fixed area (e.g., the reaction zone). The countdown starts when the microfluidic sample begins to flow, and the microfluidic sample reaches the fixed area when the countdown ends. The actuator provides pressure to the pump body of the detection device. After the microfluidic sample to be tested is successfully added, the actuator provides pressure to the pump body, compressing the pump body to drive the sample to flow to the downstream area of ​​the microfluidic channel in the detection device. The downstream area here refers to, for example, the processing zone, the reaction zone, the detection zone, etc. In some implementations, the pressure applied by the actuator to the pump body can be preset and correlated with the position of the microfluidic sample within the microfluidic channel after it enters the channel. That is, a preset pressure value applied to the pump body corresponds to a specific position of the microfluidic sample within the channel. This design can be used to confirm successful microfluidic sample addition and to determine if the microfluidic sample has entered a specific region. For example, when the actuator applies a pressure value of a certain value, the microfluidic sample flows into the reaction zone. In such an implementation, the timer does not need to be set to count down the time it takes for the microfluidic sample to reach a fixed region. The actuator provides pressure to the valve body to close the valve body, thus closing the microfluidic channel, or releases pressure to open the valve body, thus opening the microfluidic channel. The magnetic module includes a magnet and a magnetic shield. The magnet is fixed in the downstream region of the microfluidic channel in the detection device, and the magnetic shield isolates the magnet from the downstream region of the microfluidic channel. When the microfluidic sample to be tested enters the downstream region of the microfluidic channel and the microfluidic sample to be tested needs to be separated by magnetic adsorption, the main control board commands the magnetic shield to move so that the magnetic field generated by the magnet acts on the microfluidic sample to be tested.

[0054] The temperature module includes a heating unit, which is fixed in the downstream region of the microfluidic channel in the detection device. The main control board commands the heating unit to provide the required processing temperature or the required reaction temperature to the microfluidic sample to be tested.

[0055] The system consisting of the aforementioned microfluidic sample detection device and detection equipment can be used as a microfluidic sample extraction system for the separation of red blood cells and plasma in blood samples and the extraction of plasma. Red blood cells are the target, and the reagent pre-placed in the reaction zone is an antibody that binds to magnetic particles and is designed to target red blood cells.

[0056] Turn on the power to the detection equipment, place the microfluidic detection device into the load position of the microfluidic detection equipment, and expose the fluid inlet to the environment. The load position generates a load signal and transmits it to the main control board. In another embodiment, the load position does not generate a load signal; instead, the main control board receives the load signal by manually clicking a button on the detection equipment. The microfluidic detection device is connected to the microfluidic detection equipment via signal. The main control board commands the actuator to open one valve body and close the other valve body. The timer begins its first countdown. Simultaneously, a blood sample is added to the fluid inlet and enters the microfluidic channel. During the process of the blood sample entering the microfluidic channel, some gas is discharged through the fluid outlet. The volume of discharged gas is proportional to the volume of the blood sample entering. The blood sample stops entering when its tip contacts the lock body.

[0057] When the timer finishes its first countdown, the main control board confirms that the sample addition has been successful and commands the actuator to close one valve body, open the other valve body, and compress the pump body to drive the blood sample from the other valve body to the lock body into the reaction zone. The timer then begins its second countdown. During the process of the blood sample entering the reaction zone, some gas is discharged through the fluid outlet. The volume of the discharged gas is proportional to the volume of the blood sample that has entered.

[0058] When the front end of the blood sample contacts the end of the reaction zone, indicating that the reaction zone is full, the second countdown of the timer ends. The main control board commands the actuator to stop compressing the pump, and the third countdown of the timer begins. The blood sample reacts with the anti-erythrocyte antibodies bound to the magnetic particles in the reaction zone. The erythrocytes in the blood sample bind to the antibodies, forming a magnetic particle-anti-erythrocyte antibody-erythrocyte complex. The third countdown of the timer ends, and the main control board commands the magnetic shield in the magnetic module to move away from the space between the magnet and the microfluidic channel, exposing the reaction zone to the magnetic field generated by the magnet. The erythrocytes bound to the antibodies are adsorbed onto the wall of the microfluidic channel in the reaction zone due to the magnetic field. The main control board commands the actuator to continue compressing the pump to drive the plasma that has lost erythrocytes out of the reaction zone and into the collection zone. Finally, the plasma that has lost erythrocytes can flow out from the fluid outlet for subsequent use.

[0059] In another implementation, there is no first countdown; the optical module determines that the blood sample has been successfully added. In another implementation, there is no second countdown; the pressure value applied to the pump by the actuator determines that the blood sample has filled the reaction zone.

[0060] Example 2

[0061] The microfluidic sample detection system, composed of the microfluidic sample detection device and detection equipment in Example 1, was applied to the detection of urinary glucose. Glucose was used as the target analyte. The reagents pre-placed in the reaction zone 131 were prepared by combining disodium hydrogen phosphate, citric acid, polyvinylpyrrolidone, dimethyl maleate, o-toluidine, peroxidase, glucose oxidase, orange-yellow dye, enzyme stabilizer, and enzyme activator.

[0062] Turn on the power of the detection equipment, place the microfluidic detection device into the load position of the microfluidic detection equipment, and expose the fluid inlet to the environment. The load position generates a load signal and transmits it to the main control board. The microfluidic detection device is connected to the microfluidic detection equipment. The main control board commands the actuator to open one valve body and close the other valve body. The timer starts its first countdown. At the same time, add the urine sample to be tested into the fluid inlet and enter the microfluidic channel. During the process of the urine sample entering the microfluidic channel, some gas is discharged through the fluid outlet. The volume of discharged gas is proportional to the volume of the urine sample entering. When the front end of the urine sample contacts the lock body, the urine sample stops entering. The timer ends its first countdown, and the main control board receives the countdown end signal.

[0063] In another implementation, the load bit does not generate a load signal; instead, the main control board transmits the load signal by manually clicking a button on the detection device.

[0064] The main control board confirms that the sample addition has been successful and commands the actuator to close one valve body, open the other valve body, and compress the pump body to drive the urine sample to be tested from the other valve body to the lock body into the reaction zone. The timer starts its second countdown. During the process of the urine sample to be tested entering the reaction zone, some gas is discharged through the fluid outlet. The volume of the discharged gas is proportional to the volume of the urine sample to be tested entering.

[0065] When the front end of the urine sample to be tested contacts the end of the reaction zone, indicating that the reaction zone is full, the second countdown of the timer ends. The main control board then commands the actuator to stop the pump, and the third countdown of the timer begins. The urine sample reacts with the preset reagent in the reaction zone. When the third countdown of the timer ends, the main control board commands the optical detection unit in the optical detection module to detect the optical signal of the reaction result in the reaction zone. The photoelectric conversion unit converts the optical signal into an electrical signal value. The main control board receives the detected electrical signal value, determines whether glucose is present in the urine sample, and commands the display screen to show the detection result. At the same time, it commands the actuator to resume the pump to drive the waste liquid after the reaction into the collection zone. As the waste liquid enters the collection zone, some gas is discharged through the fluid outlet. The collection zone absorbs the waste liquid after the reaction. The used microfluidic sample detection device is then removed from the detection equipment and discarded.

[0066] In another implementation, there is no initial countdown; instead, successful sample addition is determined by observation. The detection device is equipped with a cover that generates a successful sample addition signal. When the urine sample to be tested is added to the fluid inlet and the front end of the sample contacts the lock and stops flowing, the cover is closed. The cover generates a successful sample addition signal and transmits it to the main control board to confirm successful addition. In another implementation, there is no initial countdown; the optical module determines successful urine sample addition. In yet another implementation, there is no second countdown; the pressure value applied to the pump by the actuator determines that the urine sample has filled the reaction zone.

[0067] Example 3

[0068] like Figure 1-2 As shown, this is a second embodiment of the microfluidic detection device of the present invention. Based on embodiment 1, it further includes a conductive system composed of electrodes. The conductive system includes two electrodes, electrode one 51 and electrode two 52. Electrode one further includes electrode contact one 511 and electrode sensing end one 512, and electrode two further includes electrode contact two 521 and electrode sensing end two 522. Electrode sensing end one 512 and electrode sensing end two 522 are located at opposite ends of the reaction zone 131, i.e., electrode sensing end one is located at the beginning of the reaction zone, and electrode sensing end two is located at the end of the reaction zone. Both electrode sensing end one and electrode sensing end two are located downstream of and close to valve body two, thereby ensuring that the pump can perform its driving function downstream of valve body two. The lock body can be located in or outside the reaction zone, but it is always located upstream of the end of the reaction zone. Figure 1-2 As shown in (a), the lock body is located in the reaction zone and downstream of the starting end of the reaction zone. That is, the lock body is located in the microfluidic channel between electrode sensing end one and electrode sensing end two, and electrode sensing end one and electrode sensing end two are also located downstream of valve body two. Figure 1-2 As shown in (b), the lock body is outside the reaction zone and close to electrode sensing end one, located in the microfluidic channel upstream of electrode sensing end one. That is to say, the lock body is not between electrode sensing end one and electrode sensing end two, but is located downstream of valve body two.

[0069] like Figure 7 The diagram shows the structural design of a load microfluidic detection device. To achieve detection functions including electrochemical methods, it includes a main control board, display screen, load position, magnetic module, temperature module, electrical module, actuator, battery, power interface, and timer. Of course, if only a certain type of detection is required, the detection device can be simplified, for example, to only include the main control board, load position, actuator, electrical module, power interface, and timer.

[0070] The power interface provides power for the instrument's operation and can charge the battery. The battery allows the testing equipment to operate normally without a power source, ensuring portability. After the testing equipment is powered on, the testing device is placed inside. When the microfluidic sample testing device is accurately placed on the load position within the testing equipment, the load position generates a load signal and transmits it to the main control board. The testing device and the testing equipment then establish a signal connection, and the main control board commands the testing equipment to begin operation. In another implementation, the load position does not generate a load signal; instead, the load signal is transmitted to the main control board by manually clicking a button on the testing device.

[0071] The electrical module includes an electrical connector and an electrical detection unit. The electrical connector provides mating electrical contacts for the electrode contacts in the microfluidic detection device. The number of electrical contacts in the microfluidic detection device corresponds to the number of electrode contacts in the microfluidic detection device. The main control board commands the electrical detection unit to output a continuous, weak electrical signal (the output stops after the microfluidic sample to be tested is successfully added). This signal cannot be detected in the event of an open circuit. When the detection device is accurately placed on the load position of the detection device, the electrode contacts on the detection device pair with the electrical contacts on the electrical connector, indicating that the detection device and the detection device have completed the electrical connection. The electrical module determines that the microfluidic sample to be tested has been successfully added by detecting the electrical signal between the electrodes. Alternatively, since the time for a microfluidic sample of a certain volume to pass through a microfluidic channel of a certain size is fixed, a sample addition waiting time can be preset in a timer, with the time when the detection device and the detection device complete the signal connection as the starting point for the sample addition waiting time. When the timer expires, it is determined that the microfluidic sample to be tested has been successfully added.

[0072] After the microfluidic sample under test completes the reaction, the electrical detection unit of the electrical module detects the current generated by the electron transfer. The current signal passes through electrode one and electrode two, and is transmitted to the main control board of the detection device through the corresponding electrical contacts of electrode one and electrode two. After processing by the main control board, the detection result is displayed on the screen.

[0073] In some implementations, a countdown timer for microfluidic sample addition can be set in the timer, with the time when the detection device and detection equipment complete the electrical connection of the electrodes serving as the starting point for the sample addition waiting time. When the timer expires, it is determined that the microfluidic sample to be tested has been successfully added.

[0074] The actuator provides pressure to the valve body to control its opening and closing. The actuator also provides pressure to the pump body of the detection device after successful addition of the microfluidic sample, compressing the pump body to drive the sample flow to the downstream region of the microfluidic channel within the detection device. This downstream region refers to, for example, the processing zone, reaction zone, or detection zone. In some embodiments, the pressure applied to the pump body by the actuator can be preset and correlated with the position of the microfluidic sample within the microfluidic channel. That is, a preset pressure value applied to the pump body corresponds to a specific position of the microfluidic sample within the microfluidic channel. This design can be used to confirm successful microfluidic sample addition and to determine that the microfluidic sample has entered a specific region. In such embodiments, a timer is not needed to determine successful addition or that the microfluidic sample has reached a fixed region.

[0075] The magnetic module includes a magnet and a magnetic shield. The magnet is fixed in the downstream region of the microfluidic channel in the detection device, and the magnetic shield isolates the magnet from the downstream region of the microfluidic channel. When the microfluidic sample to be tested enters the downstream region of the microfluidic channel and the microfluidic sample to be tested needs to be separated by magnetic adsorption, the main control board commands the magnetic shield to move so that the magnetic field generated by the magnet acts on the microfluidic sample to be tested.

[0076] The temperature module includes a heating unit, which is fixed in the downstream region of the microfluidic channel in the detection device. The main control board commands the heating unit to provide the required processing temperature or the required reaction temperature to the microfluidic sample to be tested.

[0077] The microfluidic sample detection system composed of the aforementioned microfluidic sample detection device and detection equipment can also be applied to the separation of red blood cells and plasma in blood samples, as well as the extraction of plasma. The reagent pre-placed in the reaction zone is an antibody that binds to magnetic particles and is designed to fight red blood cells.

[0078] Compared to Example 1, the difference is that after the power of the detection device is turned on, the main control board commands the electrical module to apply a weak electrical signal to the two electrical contacts corresponding to electrode one and electrode two. After the blood sample enters the microfluidic channel, the front end of the blood sample contacts electrode sensing end two. A conductive path is formed between electrode sensing end one and electrode sensing end two through the blood sample to be tested. The electrical signal can be detected through electrode contact one and electrode contact two. The main control board commands the actuator to stop compressing the pump body, and at the same time commands the electrical module to stop applying the weak signal between the two electrical contacts corresponding to electrode contact one and electrode contact two. This can replace the countdown function of a timer.

[0079] Example 4

[0080] In Example 3, the microfluidic sample detection system, consisting of a microfluidic sample detection device and a detection equipment, was applied to the detection of blood glucose. Glucose was used as the target substance, and the reagent pre-placed in the reaction zone 131 was a preparation formed by glucose dehydrogenase, an electron mediator, a surfactant, and a buffer solution.

[0081] Turn on the power to the testing equipment, place the microfluidic detection device into the load position of the microfluidic detection equipment, and pair the electrical contacts in the testing equipment with electrode contacts one and two. The load position generates a load signal and transmits it to the main control board, indicating that the detection device and the testing equipment have completed the electrode electrical connection. In another embodiment, the load position does not generate a load signal; instead, the main control board receives the load signal by manually clicking a button on the testing equipment.

[0082] When the fluid inlet is exposed to the environment, the main control board receives a signal indicating the electrical connection of the electrodes and commands the actuator to open one valve body and close the other. The timer begins its first countdown. The main control board commands the electrical module to apply a weak electrical signal between the two electrical contacts corresponding to electrodes one and two. Simultaneously, the blood sample to be tested is added to the fluid inlet and enters the microfluidic channel. During the process of the blood sample entering the microfluidic channel, some gas is discharged through the fluid outlet. The volume of the discharged gas is proportional to the volume of the blood sample entering. When the front end of the blood sample contacts the lock, the blood sample stops entering.

[0083] Once the timer finishes its first countdown, the main control board confirms that the sample addition has been successful and commands the actuator to close one valve body, open the other valve body, and compress the pump body to drive the blood sample to be tested from the other valve body to the lock body into the reaction zone. During the process of the blood sample to be tested entering the reaction zone, some gas is discharged through the fluid outlet, and the volume of the discharged gas is proportional to the volume of the blood sample to be tested entering.

[0084] The front end of the blood sample to be tested contacts the termination end of the reaction zone. A conductive path is formed between electrode sensing terminals one and two through the blood sample. An electrical signal can be detected through electrode contacts one and two, indicating that the microfluidic sample has traveled to the reaction zone. The main control board commands the actuator to stop compressing the pump body, and simultaneously commands the electrical module to stop applying a weak electrical signal between the two electrical contacts corresponding to electrode contacts one and two. The system waits for the blood sample to react with the pre-set reagent in the reaction zone and for electron transfer to occur. The current characterizing the reaction result is sensed by the electrode sensing. Terminal 1 and electrode sensing terminal 2 receive the signal and transmit it to the detection device through electrode contact 1, electrode contact 2, electrical contact 1, and electrode contact 2. The main control board receives and detects the current generated by electron transfer, processes and determines the glucose concentration in the blood sample, and commands the display screen to show the test results. At the same time, it commands the actuator to restore the compression pump body to drive the waste liquid after the reaction into the liquid collection zone. While the waste liquid after the reaction enters the liquid collection zone, some gas is discharged through the fluid outlet. The liquid collection zone absorbs the waste liquid after the reaction. The used microfluidic sample detection device is then removed from the detection device and discarded.

[0085] Example 5

[0086] like Figure 1-3 As shown, this is a third embodiment of the microfluidic detection device of the present invention. Based on embodiment 3 or any other embodiment (such as embodiment 1), it further includes a microfluidic channel branch 132, a branch outlet 121, and electrodes 53, 54, 531, 532, 541, and 542. The branch outlet is a gas outlet, which only allows gas to flow out.

[0087] The difference from other embodiments is that after the sample to be tested enters the microfluidic channel through the fluid inlet, on the one hand, the branch of the sample to be tested flows towards the lock body and stops flowing when the front end of the sample to be tested contacts the lock body; on the other hand, the branch of the sample to be tested flows towards the branch outlet and stops flowing when the front end of the sample to be tested approaches or reaches the branch outlet.

[0088] The difference from other embodiments lies in that, after the power to the detection device is turned on, the main control board commands the electrical module to apply a weak electrical signal between the two electrical contacts corresponding to electrodes three and four. The sample to be tested is connected to electrode sensing terminals three and four in the microfluidic channel branch, thus forming a circuit between electrodes three and four, confirming successful sample addition. In another embodiment, interference compensation calculation is performed by capturing the electrical signal between electrode sensing terminals three and four.

[0089] Example 6

[0090] like Figure 2As shown, this is the fifth embodiment of the microfluidic detection device of the present invention. Based on embodiment 3, the conductive system further includes electrode 55, that is, the electrode system includes three electrodes, electrode 5 including electrode contact 551 and electrode sensing end 552. Region 1 1311 and Region 2 1312 are formed in the microfluidic channel. At this time, Region 2 is the downstream region. Region 1 is a reaction zone, Region 2 is a reaction zone, or both Region 1 and Region 2 are reaction zones. Electrode sensing end 1 is located at the starting end of Region 1, electrode sensing end 2 is located at the ending end of Region 1 and is also the starting end of Region 2, and electrode sensing end 5 is located at the ending end of Region 2. Electrode sensing end 1, electrode sensing end 2, and electrode sensing end 5 are all located downstream of valve body 2, and electrode sensing end 1 is close to valve body 2, thereby ensuring that Region 1 and Region 2 are downstream of valve body 2, and maximizing the duration of pump driving force while ensuring that the pump body can perform its driving function. The lock body is located in region one and downstream of the starting end of region one, that is, the lock body is located between electrode sensing end one and electrode sensing end two and downstream of valve body two; or the lock body is located outside region one and upstream of the starting end of region one, that is, the lock body is located outside electrode sensing end one and electrode sensing end two and downstream of valve body two.

[0091] For example, red blood cells are the first target, and the reagent pre-set in region one is an antibody that binds to red blood cells with magnetic particles. Glucose is the second target, and the reagent pre-set in region two is a formulation containing glucose oxidoreductase and an electron mediator. In this case, both region one and region two are reaction zones.

[0092] Turn on the power to the detection equipment, place the microfluidic detection device into the load position of the microfluidic detection equipment, and pair the electrical contacts in the detection equipment with electrode contacts one, two, and five. The load position generates a load signal and transmits it to the main control board, indicating that the detection device and the detection equipment have completed the electrode electrical connection. In another embodiment, the load position does not generate a load signal; instead, the main control board receives the electrode electrical connection signal by manually clicking a button on the detection equipment. Upon receiving the electrode electrical connection signal, the main control board commands the actuator to open valve one and close valve two. The fluid inlet is exposed to the environment, the timer begins its first countdown, and the blood sample to be tested is added to the fluid inlet and enters the microfluidic channel. During the process of the blood sample entering the microfluidic channel, some gas is discharged through the fluid outlet. The volume of the discharged gas is proportional to the volume of the blood sample entering. The front end of the blood sample contacts the lock, and the blood sample stops entering.

[0093] Once the timer's first countdown ends, the main control board confirms that the sample addition has been successful and commands the electrical module to apply a weak electrical signal to the two electrical contacts corresponding to electrode one and electrode two. This commands the actuator to close valve one, open valve two, and compress the pump to drive the blood sample to be tested from valve two to the lock body into region one. During the process of the blood sample to be tested entering region one, some gas is discharged through the fluid outlet, and the volume of the discharged gas is proportional to the volume of the blood sample to be tested entering.

[0094] The blood sample to be tested contacts the termination point of area one, i.e., the second contact electrode sensing point. A conductive path is formed between electrode sensing points one and two through the blood sample. An electrical signal can be detected through electrode contacts one and two. The main control board commands the actuator to stop compressing the pump body, and simultaneously commands the electrical module to stop applying a weak electrical signal between the two electrical contacts corresponding to electrode contacts one and two, and to start applying a weak electrical signal between the two electrical contacts corresponding to electrode contacts two and five. The timer begins its second countdown, waiting for the blood sample to react with the anti-erythrocyte antibody reaction by binding with magnetic particles in area one. Red blood cells in the blood sample bind to antibodies, forming a magnetic particle-anti-red blood cell antibody-red blood cell complex. When the timer's second countdown ends, the main control board commands the magnetic shield in the magnetic module to move away from the space between the magnet and the microfluidic channel, exposing region one to the magnetic field generated by the magnet. Due to the magnetic field, the red blood cells bound to the antibodies are adsorbed onto the wall of the microfluidic channel in region one. The main control board commands the actuator to continue compressing the pump to drive the blood plasma that has lost red blood cells from region one into region two. During this process, some gas is discharged through the fluid outlet, and the volume of the discharged gas is equivalent to the volume of the blood plasma sample that has lost red blood cells entering region two.

[0095] The plasma front-end contact area two terminates at the terminal. A conductive path is formed between electrode sensing terminals two and five through the blood sample to be tested. An electrical signal can be detected through electrode contacts two and five. The main control board commands the actuator to stop the pump body compression and simultaneously commands the electrical module to stop applying a weak electrical signal between the two electrical contacts corresponding to electrode contacts two and five. It waits for the plasma to react with the preparation containing glucose oxidoreductase and electron mediator and for electron transfer to occur. The current is transmitted to the detection device through electrode sensing terminals two, five, two, five, two electrical contacts, and five electrode contacts. The main control board receives the detected current, processes and determines the glucose concentration in the plasma and commands the display screen to show the test result. At the same time, it commands the actuator to resume the pump body compression to drive the waste liquid after the reaction into the collection zone. While the waste liquid after the reaction enters the collection zone, some gas is discharged through the fluid outlet. The collection zone absorbs the waste liquid after the reaction. The used microfluidic sample detection device is removed from the detection device and discarded.

[0096] The above tests were repeated 10 times each using blood samples with glucose concentrations of 50 mg / dL, 100 mg / dL, 250 mg / dL, 400 mg / dL, and 550 mg / dL, respectively, and the results were obtained. A biochemical standard instrument was used as a control. The same blood samples were also tested 10 times each on existing conventional electrochemical blood glucose detection products: Brand A (high performance) and Brand B (mediocre performance), and the results are shown in Table 1. Table 1 shows that, compared with both Brand B and Brand A, the standard deviation and coefficient of variation are significantly reduced, indicating that the test results of this invention have lower data dispersion and lower repeatability among the glucose concentration measurements in the blood samples. The accuracy of the data in Table 1 was studied and plotted. Figure 9 , Figure 10 and Figure 11 As shown, the R of the present invention 2 Significantly outperformed traditional brand B products with average performance, and outperformed traditional brand R products with excellent performance. 2 This indicates that the glucose content measured in blood samples by the present invention is closer to the true value, and its accuracy is higher than that of existing products that perform well.

[0097] Table 1 Comparative Experiment Results of Example 6

[0098]

[0099] Of course, if the electrode is not present, Region 1 and Region 2 can also be formed in the microfluidic channel. The pre-placed reagent in Region 2 is replaced with anthrone reagent. The anthrone reagent reacts with glucose and changes color. The reaction result is detected by the optical module.

[0100] Example 7

[0101] like Figure 1-4 As shown, this is a fourth embodiment of the microfluidic detection device of the present invention. Based on embodiment 1 or any other embodiment, it further includes a frame electrode 501, an electrode frame area 5011, and a frame contact 5012, or only includes the electrode frame area 5011.

[0102] like Figure 3-2 As shown, this is the seventh embodiment of the microfluidic detection device of the present invention. Based on embodiment 6, it further includes a frame electrode 501, an electrode frame area 5011, and a frame contact 5012, or only includes the electrode frame area 5011.

[0103] The microfluidic detection device is formed by bonding a substrate and a coating. A frame-shaped electrode is printed on the substrate, and the electrode frame area is printed on the coating and protrudes from the coating. When the substrate and the coating are bonded together, the electrode frame area and the microfluidic channel in the substrate together form an area for the accumulation of pre-placed reagents. The two horizontally parallel sides of the electrode frame area can be used as the electrode sensing ends of the frame electrode.

[0104] Example 8

[0105] like Figure 3-1 As shown, this is a sixth embodiment of the microfluidic detection device of the present invention. Based on embodiment 6, the conductive system further includes electrode six 56, that is, the electrode system includes four electrodes, and electrode six includes electrode contact six 561 and electrode sensing end six 562. Region one 1311, region two 1312 and region three 1313 are formed in the microfluidic channel. At this time, region three is the downstream region. Among them, region one is a reaction zone, region two is a reaction zone, region three is a reaction zone, or any two of regions one, two and three are reaction zones, or regions one, two and three are all reaction zones. Electrode sensing end one is located at the beginning of region one, electrode sensing end two is located at the end of region one and is also the beginning of region two, electrode sensing end six is ​​located at the end of region two and is also the beginning of region three, and electrode sensing end five is located at the end of region three. The detection device also includes the optical module in embodiment 1.

[0106] For example, with C-reactive protein as the target, the reagent pre-placed in region one is a C-reactive protein antibody bound to magnetic particles, i.e., a C-reactive protein antibody labeled with magnetic particles. The reagent pre-placed in region two is a C-reactive protein antibody labeled with fluorescent particles. There is no pre-placed reagent in region three. In this case, regions one and two are the reaction regions, and region three is the detection region.

[0107] Turn on the power to the detection equipment, place the microfluidic detection device into the load position of the microfluidic detection equipment, and pair the electrical contacts in the detection equipment with electrode contacts one, two, five, and six. The load position generates a load signal and transmits it to the main control board, indicating that the detection device and the detection equipment have completed the electrode electrical connection. In another embodiment, the load position does not generate a load signal; instead, the main control board receives the electrode electrical connection signal by manually clicking a button on the detection equipment. Upon receiving the electrode electrical connection signal, the main control board commands the actuator to open valve one and close valve two. The fluid inlet is exposed to the environment, the timer begins its first countdown, and the blood sample to be tested is added to the fluid inlet and enters the microfluidic channel. The blood sample stops entering when its tip contacts the lock. When the timer's first countdown ends, the main control board confirms successful sample addition and commands the electrical module to apply a weak electrical signal to the two electrical contacts corresponding to electrodes one and two. This commands the actuator to close valve one, open valve two, and compress the pump to drive the blood sample from valve two to the lock into region one. The front end of the blood sample contacts the end of region one, forming a conductive path between electrode sensing terminals one and two. An electrical signal can be detected through electrode contacts one and two. The main control board commands the actuator to stop compressing the pump and simultaneously commands the electrical module to stop applying the weak electrical signal between the two electrical contacts corresponding to electrode contacts one and two, and between the two electrical contacts corresponding to electrode contacts two and six. The timer's second countdown begins, waiting for the blood sample to react with the magnetic particle-bound C-reactive protein antibody in region one. The C-reactive protein in the blood sample binds to the antibody, forming a magnetic particle-C-reactive protein antibody-C-reactive protein complex. When the second countdown of the timer ends, the main control board commands the actuator to continue compressing the pump body to drive the micro-blood sample containing the aforementioned complex from region one into region two and contact the termination end of region two. That is, a conductive path is formed between electrode sensing terminals two and six through the blood sample containing the aforementioned complex. An electrical signal can be detected through electrode contacts two and six. The main control board commands the actuator to stop compressing the pump body and simultaneously commands the electrical module to stop applying a weak electrical signal between the two electrical contacts corresponding to electrode contacts two and six, and to start applying a weak electrical signal between the two electrical contacts corresponding to electrode contacts six and five. The third countdown of the timer begins, waiting for the blood sample containing the aforementioned complex to bind with the fluorescently labeled C-reactive protein antibody to form a magnetic particle C-reactive protein antibody-C-reactive protein-fluorescently labeled C-reactive protein antibody complex, i.e., a double antibody sandwich complex.When the timer's third countdown ends, the main control board commands the actuator to resume the compression pump, driving the micro-blood sample containing the aforementioned complex to flow from region two into region three and contact the termination end of region three. A conductive path is formed between electrode sensing terminals two and six through the blood sample containing the aforementioned complex. An electrical signal can be detected through electrode contacts six and five. The main control board commands the actuator to stop the compression pump and simultaneously commands the electrical module to stop applying a weak electrical signal between the two electrical contacts corresponding to electrode contacts six and five. The main control board commands the magnetic shield in the magnetic module to move away from the space between the magnet and the microfluidic channel, exposing region three to the magnetic field generated by the magnet. Within the magnetic field range, the aforementioned magnetic particle C-reactive protein antibody-C-reactive protein-fluorescent particle-labeled C-reactive protein antibody complex is adsorbed onto the wall of the microfluidic channel in region three due to the magnetic field. The main control board commands the optical detection unit in the optical detection module to detect the fluorescence signal of the complex in region three. The main control board receives the detected fluorescence signal intensity, and the photoelectric conversion unit converts the fluorescence signal intensity into an electrical signal value. The main control board receives the detected electrical signal value, calculates the C-reactive protein content in the blood sample, and commands the display screen to display the result. At the same time, the main control board commands the actuator to resume the compression pump body to drive the waste liquid that has lost the aforementioned complex from region three into the liquid collection area.

[0108] The waste liquid is absorbed in the collection zone, and the used microfluidic sample detection device is removed from the detection equipment and discarded. In this embodiment, the reagents pre-placed in Zone 1 and Zone 2 can be exchanged.

[0109] In another embodiment, the travel area of ​​the microfluidic sample can be determined by the pressure value applied to the pump body by the actuator, which can replace the function of determining whether the microfluidic sample has reached the area corresponding to the two electrode sensing ends by applying a weak electrical signal between the two electrode sensing ends.

[0110] Example 9

[0111] Based on Example 8, it also includes an electrode seven, which includes an electrode contact seven and an electrode sensing end seven. Regions one, two, three and four are formed in the microfluidic channel. At this time, region three is the downstream region.

[0112] In other words, electrode sensing terminal five is located at both the end of region three and the beginning of region four, while electrode sensing terminal seven is located at the end of region four. In this case, region one is the pretreatment zone, regions two and three are reaction zones, and region four is the detection zone.

[0113] For example, using total vitamin D (total 25-hydroxyvitamin D) as the target, the reagent pre-set in region one is a VD dissociation solution; the reagent pre-set in region two is an antibody that binds to the first epitope via magnetic particles, i.e., a first epitope antibody labeled with magnetic particles; the reagent pre-set in region three is a second epitope antibody that can be labeled with fluorescein or fluorescent particles; and no reagent is pre-set in region four. Electrode sensing terminal five is located at the beginning of region four, and electrode sensing terminal seven is located at the end of region four.

[0114] Turn on the power to the detection equipment. Place the microfluidic detection device into the load position of the microfluidic detection equipment. The electrical contacts in the detection equipment pair with electrode contacts one, two, five, six, and seven. The load position generates a load signal and transmits it to the main control board, indicating that the detection device and the detection equipment have completed the electrode electrical connection. In another embodiment, the load position does not generate a load signal; instead, the main control board receives the electrode electrical connection signal by manually clicking a button on the detection equipment. Upon receiving the electrode electrical connection signal, the main control board commands the actuator to open valve one and close valve two. The fluid inlet is exposed to the environment, the timer begins its first countdown, and the blood sample to be tested is added to the fluid inlet and enters the microfluidic channel. The blood sample stops entering when its tip contacts the lock.

[0115] Once the timer's first countdown ends, the main control board confirms successful sample addition and commands the electrical module to apply a weak electrical signal to the two electrical contacts corresponding to electrodes one and two. This commands the actuator to close valve one, open valve two, and compress the pump to drive the blood sample from valve two to the lock into region one. The front end of the blood sample contacts the end of region one, forming a conductive path between electrode sensing terminals one and two. An electrical signal can be detected through electrode contacts one and two. The main control board commands the actuator to stop compressing the pump and simultaneously commands the electrical module to stop applying the weak electrical signal between the two electrical contacts corresponding to electrode contacts one and two. The timer's second countdown begins, waiting for the blood sample to be fully incubated and dissociated in region one by the VD dissociation solution. To fully release the bound VD in the blood sample, the main control board can also command the temperature module to heat region one before the second countdown begins. Dissociating VD from a blood sample is a specific treatment of bound VD. Only by specifically dissociating it from the blood can subsequent binding reactions be carried out to determine the total VD content in the blood. When the second countdown of the timer ends, the main control board commands the electrical module to begin applying a weak electrical signal between the two electrical contacts corresponding to electrode contact two and electrode contact six, and commands the actuator to continue compressing the pump body to drive the blood sample containing the dissociated state VD from region one to region two and contact the termination end of region two, i.e., the conductive path is formed between electrode sensing terminals two and six through the blood sample containing the dissociated state VD. The electrical signal can be detected through electrode contacts two and six. The main control board commands the actuator to stop compressing the pump body, and at the same time commands the electrical module to stop applying the weak electrical signal between the two electrical contacts corresponding to electrode contacts two and six. The third countdown of the timer begins, waiting for the blood sample containing the dissociated state VD to react with the first epitope antibody labeled by magnetic particles. The dissociated state VD in the blood sample binds to the antibody to form a magnetic particle VD antibody-VD antigen complex. When the timer's third countdown ends, the main control board commands the actuator to continue compressing the pump body to drive the micro-blood sample containing the aforementioned complex from region two into region three and contact the termination end of region three. It also commands the electrical module to apply a weak electrical signal between the two electrical contacts corresponding to electrode contacts six and five. A conductive path is formed between electrode contacts six and five through the blood sample containing the aforementioned complex, and the electrical signal can be detected through electrode contacts six and five. The main control board commands the actuator to stop compressing the pump body and simultaneously commands the electrical module to stop applying the weak electrical signal between the two electrical contacts corresponding to electrode contacts six and five. The timer's fourth countdown begins, waiting for the blood sample containing the aforementioned complex to bind with the fluorescently labeled second epitope antibody, forming a magnetic particle VD antibody-VD antigen-fluorescent particle VD antibody complex, i.e., a double-antibody sandwich complex.When the timer's fourth countdown ends, the main control board commands the actuator to resume the compression pump to drive the micro-blood sample containing the aforementioned complex from region three into region four, contacting the termination end of region four. The main control board then commands the electrical module to apply a weak electrical signal between the two electrical contacts corresponding to electrode contacts five and seven. A conductive path is formed between electrode sensing terminals five and seven through the blood sample containing the aforementioned complex, allowing the electrical signal to be detected. The main control board commands the actuator to stop the compression pump and simultaneously commands the electrical module to stop applying the weak electrical signal between the two electrical contacts corresponding to electrode contacts five and seven. The main control board then commands the magnetic shield in the magnetic module to move, thereby... Leaving the space between the magnet and the microfluidic channel, region three is exposed to the magnetic field generated by the magnet. The aforementioned magnetic particle VD antibody-VD antigen-fluorescent particle VD antibody complex is adsorbed onto the wall of the microfluidic channel in region four due to the magnetic field. The main control board commands the optical detection unit in the optical detection module to detect the fluorescence signal of the complex in region four. The main control board receives the detected fluorescence signal intensity, and the photoelectric conversion unit converts the fluorescence signal intensity into an electrical signal value. The main control board receives the detected electrical signal value, calculates the total VD content in the blood sample, and commands the display screen to display the result. At the same time, the main control board commands the actuator to restore the compression pump body to drive the waste liquid that has lost the aforementioned complex from region three into the collection area.

[0116] The waste liquid is absorbed in the collection zone, and the used microfluidic sample detection device is removed from the detection equipment and discarded. In this embodiment, the reagents pre-placed in Zones 2 and 3 can be exchanged.

[0117] In another embodiment, the travel area of ​​the microfluidic sample can be determined by the pressure value applied to the pump body by the actuator, which can replace the function of determining whether the microfluidic sample has reached the area corresponding to the two electrode sensing ends by applying a weak electrical signal between the two electrode sensing ends.

[0118] Example 10

[0119] like Figure 4-1 As shown, this is the eighth embodiment of the microfluidic detection device of the present invention. In embodiment 1 or any other embodiment, it includes electrode 1 51, electrode contact 1 511, electrode sensing end 1 512, electrode 8 58, electrode contact 8 581, and electrode sensing end 8 582. Electrode sensing end 8 is located upstream of valve body 2, and electrode sensing end 1 is located downstream of electrode sensing end 8. Electrode sensing end 1, electrode sensing end 8, and the microfluidic channel together form a start-up area. The start-up area is close to the microfluidic inlet. Electrode sensing end 8 is the starting end of the start-up area, and electrode sensing end 1 is the ending end of the start-up area. The existence of the start-up area can replace the function of the timer's first countdown before successful sample addition.

[0120] like Figure 4-2As shown, this is the ninth embodiment of the microfluidic detection device of the present invention. Figure 4-1 Based on this, it also includes electrode two, electrode contact two, and electrode sensing end two. The lock body is located in the microfluidic channel between electrode sensing end one and electrode sensing end two, and is downstream of electrode sensing end one. Electrode sensing end one and electrode sensing end two are also located downstream of valve body two, close to valve body two. In another embodiment, refer to... Figure 1-2 (b), distinct from Figure 4-2 In this configuration, the lock body is not located in the microfluidic channel between electrode sensing end one and electrode sensing end two, but rather in the microfluidic channel between electrode sensing end one and electrode sensing end eight. The lock body is located upstream of electrode sensing end one, while electrode sensing end one and electrode sensing end two are still close to valve body two and are also located downstream of valve body two.

[0121] The difference from other embodiments is that the detection device includes electrical contacts that pair with electrode contact one and electrode contact eight. While the main control board commands the actuator to open valve body one and close valve body two, it also commands the electrical module to apply a weak electrical signal to electrode body one and electrode body eight through the contacts. When the tip of the microfluidic sample contacts electrode sensing end one, a conductive path is formed between electrode sensing end one and electrode sensing end eight through the microfluidic sample. The electrical signal can be detected through electrode contact one and electrode contact eight. The main control board determines that the microfluidic sample has been successfully added and commands valve body one to close and valve body two to open. The microfluidic sample in the microfluidic channel between valve body two and electrode sensing end one is a determined microfluidic sample volume, thus providing a double guarantee for successful sample addition together with the lock body.

[0122] Example 11

[0123] The microfluidic detection device of the present invention can be combined with the devices in Examples 1 to 10 and applied to the combined detection of multiple indicators. Pregnant women are at risk of gestational diabetes and anemia, so it is necessary to monitor blood glucose, beta-hydroxybutyrate and hemoglobin during pregnancy.

[0124] like Figure 5 As shown, an apparatus for simultaneously detecting glucose, β-hydroxybutyric acid and hemoglobin in blood is provided. Apparatus unit 101 is used to detect glucose, apparatus unit 2 102 is used to detect β-hydroxybutyric acid, and apparatus units 3 104 and 4 105 are used to detect hemoglobin.

[0125] like Figure 8 As shown, the microfluidic detection device includes both optical and electrical modules.

[0126] To amplify the signal response of β-hydroxybutyric acid, the sensing end 1021 of the second electrode in unit two is thickened. To ensure that the area where the sample remains and the area that reacts with the pre-placed reagent coincide, the horizontal side of the first electrode frame area 1032 in unit three is aligned with the sensing end 1031 of the third electrode, and the horizontal side of the second electrode frame area 1042 in unit four is aligned with the sensing end 1041 of the fourth electrode.

[0127] Each of the four device units consists of two regions formed by the electrode sensing ends of three electrodes. The reagents pre-placed in the two regions of device unit one, from bottom to top, are: first, an antibody that binds magnetic particles to anti-erythrocytes; second, a formulation containing glucose oxidoreductase and an electron mediator. Similarly, the reagents pre-placed in the two regions of device unit two, from bottom to top, are: first, an antibody that binds magnetic particles to anti-erythrocytes; second, a formulation containing β-hydroxybutyrate oxidoreductase and an electron mediator. The reagents pre-placed in the two regions of device units three and four, from bottom to top, are: first, a hemoglobin antibody bound to magnetic particles; second, a hemoglobin antibody labeled with fluorescent particles. Device unit four serves as a control channel, and device unit three serves as a detection channel, thus eliminating errors through the control channel while detecting hemoglobin levels.

[0128] The detection processes of Unit 1 and Unit 2 are the same as in Example 6. The detection processes of Unit 3 and Unit 4 are similar to those in Example 8, except that after the formation of the dual-antibody sandwich complex and the third countdown of the timer, the main control board commands the magnetic shield in the magnetic module to move away from the space between the magnet and the microfluidic channel, exposing Region 2 to the magnetic field generated by the magnet. The aforementioned dual-antibody sandwich complex is adsorbed onto the wall of the microfluidic channel in Region 2 due to the magnetic field. The main control board commands the optical detection unit in the optical detection module to detect the fluorescence signal of the complex in Region 2. The main control board receives the detected fluorescence signal intensity, and the photoelectric conversion unit converts the fluorescence signal intensity into an electrical signal value. The main control board receives the detected electrical signal value, calculates the hemoglobin content in the blood sample, and commands the display screen to display the result. At the same time, the main control board commands the actuator to restore the compression pump body to drive the waste liquid that has lost the aforementioned complex from Region 2 into the collection area.

[0129] The above embodiments are only used to illustrate the technical solutions of the present invention more clearly, and are therefore only examples and should not be used to limit the scope of protection of the present invention.

[0130] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains; the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the invention; the terms “comprising” and “having”, and any variations thereof, in the specification and claims of this invention are intended to cover non-exclusive inclusion.

[0131] In the description of this invention, technical terms such as "first," "second," "third," and "fourth" are used only to distinguish different objects and should not be construed as indicating or implying relative importance or implicitly specifying the number, specific order, or primary and secondary relationship of the indicated technical features. In the description of the embodiments of this invention, "a plurality of" means two or more, unless otherwise explicitly defined.

[0132] In this document, the terms "embodiment" and "implementation" mean that a particular feature, structure, or characteristic described in connection with an embodiment may be included in at least one embodiment of the invention. The appearance of this phrase in various places in the specification does not necessarily refer to the same embodiment, nor is it a separate or alternative embodiment that is mutually exclusive with other embodiments.

[0133] The above embodiments are merely illustrative of the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features therein. These modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention, and they should all be covered within the scope of the claims and specification of the present invention. In particular, as long as there is no structural conflict, the various technical features mentioned in the embodiments can be combined in any way. The present invention is not limited to the specific embodiments disclosed herein, but includes all technical solutions falling within the scope of the claims.

Claims

1. A microfluidic sample detection device, characterized in that, This includes fluid inlet, microfluidic channels, and gas outlet, as well as: The reaction zone is located in the microfluidic channel between the fluid inlet and the gas outlet; the liquid collection zone is located downstream of the reaction zone, and the gas outlet is the outlet of the liquid collection zone. The first pump body, located upstream of the reaction zone, is compressed when communicating with the microfluidic channel to provide driving force for the flow of the microfluidic sample. The volume of the microfluidic sample that propels forward after entering the microfluidic channel is proportional to the volume of gas discharged from the gas outlet at the same time. The pressure value applied to the first pump body is preset and associated with the travel position of the microfluidic sample in the microfluidic channel after entering it. A first valve body is located downstream of the fluid inlet and controls the opening and closing of the microfluidic channel and the fluid inlet; The second valve body is located downstream of the first valve body and independently controls the opening and closing of the first pump body and the microfluidic channel. The first valve body and the second valve body have opposite working states. The first locking body is located downstream of the second valve body and upstream of the reaction zone termination end. Together with the second valve body, it defines the volume of the microfluidic sample. When the microfluidic sample to be tested enters the microfluidic channel from the fluid inlet and the front end of the microfluidic sample to be tested contacts the first locking body, the flow stops. After the first valve body is closed, the second valve body is opened, and the first pump body drives the microfluidic sample to be tested between the second valve body and the first locking body to flow downstream in the microfluidic channel. It also includes a conductive system composed of electrodes, the conductive system comprising at least two electrodes, the electrode sensing ends being located at the two ends of the reaction zone respectively, and the electrode contacts on the microfluidic sample detection device being paired with the electrical contacts on the microfluidic detection device.

2. The microfluidic sample detection device according to claim 1, characterized in that, The microfluidic channel includes a first region and a second region, at least the first region or the second region is a reaction zone, and the first pump can drive the microfluidic sample to enter the first region and the second region in sequence.

3. The microfluidic sample detection device according to claim 2, characterized in that, The microfluidic channel includes a first region, a second region, and a third region, at least the first region, the second region, or the third region is a reaction zone, and the first pump can drive the microfluidic sample to enter the first region, the second region, and the third region in sequence.

4. The microfluidic sample detection device according to claim 1, characterized in that, The conductive system includes at least three electrodes. The first electrode sensing end and the second electrode sensing end are distributed at the beginning and end of the first region, and the second electrode sensing end and the third electrode sensing end are distributed at the beginning and end of the second region. At least the first region or the second region is a reaction zone. The first pump can drive the microfluidic sample to enter the first region and the second region in sequence.

5. A microfluidic sample detection device according to claim 4, characterized in that, The conductive system includes at least four electrodes. The first electrode sensing end and the second electrode sensing end are distributed at the start and end of the first region. The second electrode sensing end and the third electrode sensing end are distributed at the start and end of the second region. The third electrode sensing end and the fourth electrode sensing end are distributed at the start and end of the third region. At least the first region, the second region, or the third region is a reaction zone. The first pump can drive the microfluidic sample to enter the first region, the second region, and the third region in sequence.

6. A microfluidic sample detection device according to claim 4 or 5, characterized in that, It also includes a start electrode, the sensing end of which is located upstream of the second valve body and together with the sensing end of the first electrode downstream of the second valve body, forms a start area in the microfluidic channel. In response to the start signal generated when the microfluidic sample flows into the start area and contacts the sensing end of the first electrode, the first valve body closes and the second valve body opens.

7. A microfluidic sample detection device according to claim 4 or 5, characterized in that, The first lock body is located in the microfluidic channel between the first electrode sensing end and the second electrode sensing end, and the first electrode sensing end and the second electrode sensing end are close to the second valve body and located downstream of the second valve body.

8. A microfluidic sample detection device according to claim 6, characterized in that, The first lock body is located between the first electrode sensing end and the second electrode sensing end, and the first electrode sensing end and the second electrode sensing end are close to the second valve body and located downstream of the second valve body.

9. A microfluidic sample detection device according to claim 1, 2, or 4, characterized in that, The reaction zone is pre-positioned with an antibody that binds to magnetic particles and is designed to specifically bind to red blood cells in a blood sample.

10. A microfluidic sample detection device according to any one of claims 1-5, characterized in that, The reaction zone is pre-positioned with target antibody labeled with magnetic particles, the antibody being designed to specifically bind to the target in the sample.

11. A microfluidic sample detection device according to any one of claims 1-5, characterized in that, The reaction zone is pre-contained with fluorescently labeled antibodies, which are designed to specifically bind to the target analyte in the sample.

12. A microfluidic sample detection device according to claim 2, 3, 4, or 5, characterized in that, It includes two reaction zones, each pre-positioned with a first antibody labeled with magnetic particles and a second antibody labeled with fluorescent particles. The first and second antibodies are designed to specifically bind to the target in the sample. After the target in the microfluidic sample binds to the first and second antibodies, a double-antibody sandwich complex is formed.

13. A microfluidic sample detection device according to claim 12, characterized in that, The microfluidic sample is specifically treated before it binds to the first antibody and the second antibody.

14. A microfluidic sample detection system, comprising a detection device and a microfluidic sample detection apparatus as described in any one of claims 1-13, wherein the microfluidic sample detection apparatus is mounted in the detection device, and the detection device is electrically or signalally connected to the electrodes of the microfluidic sample detection apparatus, thereby controlling the opening or closing of the first valve body and the second valve body, and controlling the compression or release of the first pump body via an actuator.

15. A microfluidic sample detection system according to claim 14, characterized in that, The microfluidic sample detection device is electrically connected to the electrodes of the detection equipment via electrical contacts.

Images