A microfluidic sample testing method

Abstract

The application provides a microfluid sample detection method, first, opening a microfluid detection device and placing a microfluid detection device into a loading position, the microfluid detection device is signal connected with the microfluid detection device; second, an actuator keeps a first valve body open and a second valve body closed, a microfluid sample enters a microfluid channel through a fluid inlet, part of gas is discharged through a fluid outlet, until the front end of the microfluid sample contacts a first lock body and stops flowing; then, a main control board determines that sample adding is successful, the actuator closes the first valve body, opens the second valve body, and compresses a first pump body to drive the microfluid sample into a reaction area; finally, the front end of the microfluid sample contacts a reaction area terminal end, the actuator stops compressing the first pump body, waits for the countdown in a timer to end, the actuator resumes compressing the first pump body, drives the microfluid sample after reaction to continue flowing and far away from the reaction area into a liquid collection area, and a detection module detects a reaction result completion signal.

Description

Technical Field

[0001] This invention relates to the field of medical devices, and more particularly to a method for detecting microfluidic samples. 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 above problems, the present invention provides a microfluidic sample detection method, comprising the following steps: opening the microfluidic detection device and placing the microfluidic detection device in the load position, and connecting the microfluidic detection device to the microfluidic detection equipment via signal connection;

[0006] The actuator keeps the first valve body open and the second valve body closed. The microfluidic sample enters the microfluidic channel through the fluid inlet and some gas is discharged through the fluid outlet until the front end of the microfluidic sample contacts the first locking body and stops flowing.

[0007] Once the main control board confirms successful sample addition, the actuator closes the first valve, opens the second valve, and then compresses the first pump to drive the microfluidic sample into the reaction zone.

[0008] When the front end of the microfluidic sample contacts the end of the reaction zone, the actuator stops compressing the first pump body. After the countdown in the timer ends, the actuator resumes compressing the first pump body, driving the microfluidic sample after the reaction to continue flowing away from the reaction zone and enter the liquid collection zone. The detection module completes signal detection of the reaction result.

[0009] Specifically, the photodetector unit determines the travel region of the microfluidic sample by continuously detecting the light reflection state. The travel region includes the region where the sample is successfully added, the region where the microfluidic sample is processed, the region where the microfluidic sample reacts, and the region where the microfluidic sample is detected.

[0010] Specifically, the travel region of the microfluidic sample is determined by the pressure value applied to the first pump body by the actuator. The travel region includes the region where the sample is successfully added, the region where the microfluidic sample is processed, the region where the microfluidic sample reacts, and the region where the microfluidic sample is detected.

[0011] Furthermore, the microfluidic detection device is turned on and placed in the load position. The first electrode contact and the second electrode contact are paired with the electrical contacts, and the electrodes of the microfluidic detection device and the microfluidic detection equipment are electrically connected.

[0012] Specifically, the electrical module determines the travel region of the microfluidic sample through the first and second electrode sensing terminals. The travel region includes the region where the sample is successfully added, the region where the microfluidic sample is processed, the region where the microfluidic sample reacts, and the region where the microfluidic sample is detected.

[0013] Furthermore, after the countdown in the timer ends, the electrical module receives the reaction result signal through the first electrode sensing terminal and the second electrode sensing terminal, and transmits it to the microfluidic detection device through the first electrode contact, the second electrode contact, and the electrical contacts.

[0014] In another embodiment, the actuator compresses the first pump body to drive the microfluidic sample into the first region and then into the second region, with the countdown occurring before the microfluidic sample enters the second region.

[0015] Furthermore, the first electrode contact, the second electrode contact, and the third electrode contact are respectively paired with electrical contacts to achieve electrical connection of the electrodes of the microfluidic detection device and the microfluidic detection equipment.

[0016] Furthermore, the electrical module determines the travel region of the microfluidic sample through at least two of the first electrode sensing terminal, the second electrode sensing terminal, and the third electrode sensing terminal. The travel region includes the region where the sample is successfully added, the region where the microfluidic sample is processed, the region where the microfluidic sample reacts, and the region where the microfluidic sample is detected.

[0017] Specifically, the countdown also occurs after the microfluidic sample enters the second region. After the countdown ends after the microfluidic sample enters the second region, the electrical module receives the reaction result signal through the second electrode sensing end and the third electrode sensing end, and transmits it to the microfluidic detection device through the second electrode contact, the third electrode contact, and the electrical contacts.

[0018] In another embodiment, the microfluidic sample enters the first region, the second region, and the third region sequentially, with the countdown occurring before the microfluidic sample enters the third region.

[0019] Furthermore, the first electrode contact, the second electrode contact, the third electrode contact, and the fourth electrode contact are respectively paired with electrical contacts to achieve electrical connection of the electrodes of the microfluidic detection device and the microfluidic detection equipment.

[0020] Furthermore, the electrical module determines the travel region of the microfluidic sample through at least two of the first, second, third, and fourth electrode sensing terminals. The travel region includes the region where the sample is successfully added, the region where the microfluidic sample is processed, the region where the microfluidic sample reacts, and the region where the microfluidic sample is detected.

[0021] Specifically, the countdown also occurs after the microfluidic sample enters the third region. After the countdown ends after the microfluidic sample enters the third region, the electrical module receives the reaction result signal through the third electrode sensing end and the fourth electrode sensing end, and transmits it to the microfluidic detection device through the third electrode contact, the fourth electrode contact, and the electrical contacts.

[0022] In another embodiment, the microfluidic sample enters the first region, the second region, the third region, and the fourth region sequentially, with the countdown occurring before the microfluidic sample enters the fourth region.

[0023] Furthermore, the first electrode contact, the second electrode contact, the third electrode contact, the fourth electrode contact, and the fifth electrode contact are respectively paired with electrical contacts to achieve electrical connection of the electrodes of the microfluidic detection device and the microfluidic detection equipment.

[0024] Furthermore, the electrical module determines the travel region of the microfluidic sample through at least two of the first, second, third, fourth, and fifth electrode sensing terminals. The travel region includes the region where the sample is successfully added, the region where the microfluidic sample is processed, the region where the microfluidic sample reacts, and the region where the microfluidic sample is detected.

[0025] Specifically, the countdown also occurs after the microfluidic sample enters the fourth region. After the countdown ends after the microfluidic sample enters the fourth region, the electrical module receives the reaction result signal through the fourth electrode sensing end and the fifth electrode sensing end, and transmits it to the microfluidic detection device through the fourth electrode contact, the fifth electrode contact, and the electrical contacts.

[0026] In some implementations, the optical module obtains the reaction result signal by detecting the downstream region.

[0027] In some implementations, the reaction zone includes a target antibody labeled with magnetic particles, which specifically binds to the target in the microfluidic sample.

[0028] In some embodiments, the reaction zone includes fluorescently labeled antibodies that specifically bind to a target in the microfluidic sample.

[0029] In a specific implementation method

[0030] The reaction zone includes a first reaction zone and a second reaction zone, in which a magnetically labeled target antibody and a fluorescently labeled target antibody are pre-placed, respectively, in the first and second reaction zones.

[0031] The actuator compresses the first pump body to drive the microfluidic sample into the first reaction zone and then into the second reaction zone, where the target analyte, the first antibody, and the second antibody together form a bi-antibody sandwich complex.

[0032] Before the actuator compresses the first pump body and drives the reacted microfluidic sample to continue flowing, the magnetic module generates a magnetic field to adsorb the dual-antibody sandwich composite onto the wall of the microfluidic channel.

[0033] The beneficial effects of this invention include: 1. It can be applied to in vitro diagnostic products using various methodologies, and can perform single-item detection, multi-item combined detection, and even combine electrochemical detection and photochemical detection methods into the same detection process, simultaneously detecting microfluidic samples entering from a single fluid inlet; 2. The microfluidic sample after reaction enters the downstream liquid collection zone, avoiding the problem of contaminating the detection equipment; 3. During the microfluidic sample detection process, sample quantification is completed by controlling the cooperation of the valve body and the lock body, and the accuracy and repeatability of the detection results of the in vitro diagnostic electrochemical product are superior to those of traditional in vitro diagnostic electrochemical products. Attached Figure Description

[0034] Figure 1-1 One embodiment of the microfluidic detection device of the present invention.

[0035] Figure 1-2 The second embodiment of the microfluidic detection device of the present invention.

[0036] Figure 1-3 The third embodiment of the microfluidic detection device of the present invention.

[0037] Figure 1-4 The fourth embodiment of the microfluidic detection device of the present invention.

[0038] Figure 2 The fifth embodiment of the microfluidic detection device of the present invention.

[0039] Figure 3-1 The sixth embodiment of the microfluidic detection device of the present invention.

[0040] Figure 3-2 The seventh embodiment of the microfluidic detection device of the present invention.

[0041] Figure 4-1 The eighth embodiment of the microfluidic detection device of the present invention.

[0042] Figure 4-2 The ninth embodiment of the microfluidic detection device of the present invention.

[0043] Figure 5 This invention provides a combined implementation of a microfluidic detection device.

[0044] Figure 6 One of the schematic diagrams of the microfluidic detection device module of the present invention.

[0045] Figure 7 The second schematic diagram of the microfluidic detection device module of the present invention.

[0046] Figure 8 The third schematic diagram of the microfluidic detection device module of the present invention.

[0047] Figure 9Example 6: Results of the accuracy study of the product of the present invention.

[0048] Figure 10 Example 6A: Results of a study on the accuracy of traditional products.

[0049] Figure 11 Example 6B: Results of a study on the accuracy of traditional products. Detailed Implementation

[0050] 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.

[0051] Microfluidic detection device

[0052] like Figure 1-1 As shown, one embodiment of the microfluidic detection device of the present invention includes 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 in the microfluidic channel. The volume of the microfluidic sample propelled forward in the microfluidic channel is the same as the volume of gas simultaneously discharged from the fluid outlet. The liquid collection zone 14 is downstream of the microfluidic channel and also downstream of the reaction zone, used to receive the microfluidic sample after the reaction. The fluid outlet is located at the outlet of the liquid collection zone and is in communication with the environment. The environment refers to the external environment of the microfluidic detection device of the present invention, such as air or vacuum. It can be the environment set in the system of the present invention or the same as the environment outside the system of the present invention. The fluid outlet allows either gas or liquid to flow out. In most embodiments, the fluid outlet is a gas outlet, allowing only gas to flow out and preventing liquid from flowing out. In a few embodiments, the fluid outlet is both a gas and liquid outlet, allowing both gas and liquid to flow out. 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. In this case, the reaction zone is the downstream region.

[0053] 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.

[0054] 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.

[0055] 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.

[0056] 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.

[0057] 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.

[0058] like Figure 1-2 As shown, this is a second embodiment of the microfluidic detection device of the present invention. Based on the first microfluidic detection device, 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 the second valve body, thereby ensuring that the pump body can perform its driving function downstream of the second valve body. 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.

[0059] like Figure 1-3 As shown, to implement the third microfluidic detection device of the present invention, based on the first microfluidic detection device, the second microfluidic detection device, or other subsequent microfluidic detection devices, it further includes a microfluidic channel branch 132, a branch outlet 121, and electrodes 53, 54, 531, 532, 541, and 542, wherein the branch outlet is a gas outlet, allowing only gas to flow out.

[0060] like Figure 1-4 As shown, a fourth microfluidic detection device for implementing the present invention further includes a frame-shaped electrode 501, an electrode frame-shaped region 5011, and a frame-shaped contact 5012, based on the first, second, third, or other subsequent microfluidic detection devices. Alternatively, based on the first, second, third, or other subsequent microfluidic detection devices, only the electrode frame-shaped region 5011 is included.

[0061] like Figure 2 As shown, this is the fifth microfluidic detection device for implementing the present invention. Based on the second microfluidic detection device, the conductive system further includes electrode five 55, that is, the electrode system includes three electrodes, electrode five including electrode contact five 551 and electrode sensing end five 552. Region one 1311 and region two 1312 are formed in the microfluidic channel. At this time, region two is the downstream region. Region one is a reaction zone, region two is a reaction zone, or both regions one and two are reaction zones. Electrode sensing end one is located at the starting end of region one, electrode sensing end two is located at the ending end of region one and is also the starting end of region two, and electrode sensing end five is located at the ending end of region two. Electrode sensing end one, electrode sensing end two, and electrode sensing end five are all located downstream of valve body two, and electrode sensing end one is close to valve body two, thereby ensuring that region one and region two are downstream of valve body two, 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.

[0062] like Figure 3-1As shown, this is the sixth microfluidic detection device for implementing the present invention. Based on the fifth microfluidic detection device, the conductive system further includes an electrode six 56, meaning the electrode system includes four electrodes. Electrode six includes an electrode contact six 561 and an electrode sensing end six 562. Region one 1311, region two 1312, and region three 1313 are formed in the microfluidic channel, with region three being the downstream region. Region one is a reaction zone, or region two is a reaction zone, or region three is a reaction zone, or any two of regions one, two, and three are reaction zones, or all three are 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. Furthermore, it also includes electrode seven, which comprises electrode contact seven and electrode sensing end seven. Regions one, two, three, and four are formed within the microfluidic channel. That is, electrode sensing end five is located both at the end of region three and the beginning of region four, while electrode sensing end seven is located at the end of region four. In this case, region four is the most downstream region. Specifically, region one is a reaction zone, or region two is a reaction zone, or region three is a reaction zone, or region four is a reaction zone, or any two or three of regions one, two, three, and four are reaction zones, or all four regions are reaction zones.

[0063] like Figure 3-2 As shown, this is a seventh microfluidic detection device implementing the present invention. Based on the fifth microfluidic detection device, it further includes a frame electrode 501, an electrode frame region 5011, and a frame contact 5012, or only includes the electrode frame region 5011. The microfluidic detection device is formed by bonding a substrate and a coating. The frame electrode is printed on the substrate, and the electrode frame region is printed on the coating and protrudes from the coating. When the substrate and the coating are bonded together, the electrode frame region and the microfluidic channels in the substrate together form an area for the accumulation of pre-placed reagents. The two horizontally parallel sides of the electrode frame region can serve as the electrode sensing ends of the frame electrode.

[0064] like Figure 4-1 As shown, this is the eighth microfluidic detection device for implementing the present invention. Based on any one of the first to seventh microfluidic detection devices or other subsequent microfluidic detection devices, it includes an electrode 51, an electrode contact 511, an electrode sensing end 512, an electrode 58, an electrode contact 581, and an electrode sensing end 582. The electrode sensing end 58 is located upstream of the valve body 2, and the electrode sensing end 1 is located downstream of the electrode sensing end 58. The electrode sensing end 1, the electrode sensing end 58, and the microfluidic channel together form a starting area. The starting area is close to the microfluidic inlet. The electrode sensing end 8 is the starting end of the starting area, and the electrode sensing end 1 is the ending end of the starting area.

[0065] like Figure 4-2 As shown, this is the ninth microfluidic detection device for implementing 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 The lock body is not in the microfluidic channel between electrode sensing end one and electrode sensing end two, but 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.

[0066] Microfluidic detection devices one through eight can be arbitrarily copied and combined to form new microfluidic detection devices, with the combined microfluidic detection devices retaining only one fluid inlet. For example, microfluidic detection device one can be copied and combined to form a microfluidic detection device that includes one of the two microfluidic detection devices but retains only one fluid inlet, or microfluidic detection devices five and seven can both be copied and combined to form a microfluidic detection device that includes two units of microfluidic detection device five and two units of microfluidic detection device seven but retains only one fluid inlet.

[0067] like Figure 5 As shown, in order to implement the combined microfluidic detection device of the present invention, the fifth microfluidic detection device, the third microfluidic detection device based on the fifth microfluidic detection device, and the two seventh microfluidic detection devices are combined to form a microfluidic detection device that retains only one fluid inlet.

[0068] Microfluidic detection equipment

[0069] like Figure 6 , Figure 7 , Figure 8 The diagram shown illustrates the structure of a microfluidic detection device for implementing the present invention, including a main control board, a display screen, a load position, a magnetic module, a temperature module, an optical module, an electrical module, an actuator, a battery, a power interface, and a timer. In specific embodiments, at least one of the optical module and the electrical module is included; the magnetic module, temperature module, and display screen are optional designs and may or may not be included in the microfluidic detection device.

[0070] 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.

[0071] 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 in the timer, with the start point of the addition waiting time being the time when the detection device and the detection equipment complete the signal connection. When the timer expires, it is determined that the microfluidic sample to be tested has been successfully added. When the timer is used to determine successful addition, it is not necessary to use the aforementioned optical detection unit to determine successful addition. In another embodiment, a countdown can also be set in the timer to determine 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.

[0072] 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 it to drive the sample to flow to the downstream region of the microfluidic channel in the detection device. This downstream region refers to, for example, the processing zone, reaction zone, or detection zone. In some embodiments, the pressure value applied to the pump body by the actuator can be preset and correlated with the travel position of the microfluidic sample within the microfluidic channel. That is, a preset pressure value applied to the pump body corresponds to a certain position of the microfluidic sample in 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. For example, when the actuator applies pressure to a certain value, the microfluidic sample flows to the reaction zone. In such an embodiment, 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 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.

[0073] 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.

[0074] 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.

[0075] 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.

[0076] 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.

[0077] 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.

[0078] Microfluidic sample extraction system

[0079] The system formed by combining the above-mentioned microfluidic sample detection device and microfluidic sample detection equipment is applied to the separation and extraction of microfluidic samples.

[0080] Example 1

[0081] The present invention is implemented on a microfluidic sample detection system composed of one of the microfluidic detection devices and the microfluidic detection equipment. The microfluidic sample detection method of the present invention is used to detect urinary glucose, with glucose as the target. The reagent pre-placed in the reaction zone is prepared by disodium hydrogen phosphate, citric acid, polyvinylpyrrolidone, dimethyl maleate, o-toluidine, peroxidase, glucose oxidase, orange-yellow dye, enzyme stabilizer, and enzyme activator.

[0082] 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.

[0083] 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.

[0084] 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.

[0085] 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 pre-set 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 to continue flowing and enter the collection zone. As 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 then removed from the detection equipment and discarded.

[0086] In another embodiment, 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 tip of the sample is observed to contact the lock and stop flowing, the cover is closed, and the cover generates a successful sample addition signal, which is transmitted to the main control board to confirm successful addition. In another embodiment, there is no initial countdown; the optical module determines successful urine sample addition. In another embodiment, 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. In yet another embodiment, no initial countdown is required; 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 tip of the sample is observed to contact the lock and stop flowing, the cover is closed, and the cover generates a successful sample addition signal, which is transmitted to the main control board.

[0087] Example 2

[0088] The present invention is implemented on a microfluidic sample detection system composed of a second microfluidic detection device and a microfluidic detection equipment. Blood glucose is detected using the microfluidic sample detection method of the present invention, with glucose as the target. The reagent pre-placed in the reaction zone 131 is a preparation formed by glucose dehydrogenase, an electron mediator, a surfactant, and a buffer solution.

[0089] 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. Upon receiving the electrode electrical connection signal, the main control board commands the actuator to open valve body one and close valve body two. In another embodiment, 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 equipment.

[0090] When the fluid inlet is exposed to the environment, the main control board receives a signal indicating the electrical connection of the electrodes. This signal commands the actuator to open one valve body and close the other valve body. The electrical module then applies a weak electrical signal between the two electrical contacts corresponding to electrodes one and two. The timer begins its first countdown. 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.

[0091] 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.

[0092] 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.

[0093] Example 3

[0094] The present invention is implemented on a microfluidic sample detection system composed of a third microfluidic detection device and a microfluidic detection equipment.

[0095] 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.

[0096] 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.

[0097] Example 4

[0098] The present invention is implemented on a microfluidic sample detection system composed of a fourth microfluidic detection device and a microfluidic detection equipment.

[0099] The difference from other embodiments lies in that a pre-prepared liquid reagent is contained within a microfluidic channel within the electrode frame area. When a microfluidic sample flows into the electrode frame area, it reacts with the pre-prepared liquid reagent therein. Furthermore, the pre-prepared liquid reagent, contained within a microfluidic channel within the electrode frame area, reacts with the pre-prepared liquid reagent when a microfluidic sample flows into the electrode frame area, generating a signal change between the frame electrode and another electrode within the electrode frame area. This signal is then transmitted to the microfluidic detection instrument via the frame contact and the contact of the other electrode.

[0100] Example 5

[0101] The present invention is implemented on a microfluidic sample detection system composed of a microfluidic detection device and a microfluidic detection equipment.

[0102] Red blood cells are the primary target, and the reagent pre-set in region one is an antibody that binds to red blood cells with magnetic particles. Glucose is the secondary target, and the reagent pre-set in region two is a formulation containing glucose oxidoreductase and an electron mediator. In this case, both regions one and two are reaction zones.

[0103] 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, 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 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 electrode electrical connection signal by manually clicking a button on the testing equipment.

[0104] Upon receiving the signal indicating the electrical connection of the electrodes, the main control board commands the actuator to open one valve body and close the other. With the fluid inlet exposed to the environment, the main control board commands the electrical module to apply a weak electrical signal to the two electrical contacts corresponding to electrodes one and two. The timer begins its first countdown, and 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. The blood sample stops entering when its tip contacts the locking mechanism.

[0105] 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 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 area one. During the process of the blood sample to be tested entering area one, some gas is discharged through the fluid outlet. The volume of the discharged gas is proportional to the volume of the blood sample to be tested entering.

[0106] 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. The timer begins its second countdown, waiting for the blood sample to react with the anti-erythrocyte antibody in area one. The erythrocytes in the blood sample bind with the antibody, forming a magnetic particle-anti-erythrocyte antibody-erythrocyte complex. When the 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. Red blood cells bound to antibodies are adsorbed onto the wall of the microfluidic channel in region one due to the magnetic field. The main control board commands the actuator to continue compressing the pump to drive the plasma that has lost red blood cells from region one into region two. It also commands the electrical module to apply a weak electrical signal between the two electrical contacts corresponding to electrode contacts two and five. During the process, some gas is discharged through the fluid outlet, and the volume of the discharged gas is equivalent to the volume of the plasma sample that has lost red blood cells and entered region two.

[0107] 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.

[0108] 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.

[0109] Table 1 Comparative Experiment Results of Example 6

[0110]

[0111] Example 6

[0112] The present invention is implemented on a microfluidic sample detection system composed of a microfluidic detection device and a microfluidic detection equipment.

[0113] Using 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 areas, and region three is the detection area.

[0114] 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 main control board commands the electrical module to apply a weak electrical signal to the two electrical contacts corresponding to electrodes one and two, starting the first countdown of the timer. Simultaneously, the blood sample to be tested is added to the fluid inlet and enters the microfluidic channel. The front end of the blood sample contacts the lock, stopping the blood sample from entering. 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 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 area one.

[0115] The blood sample to be tested contacts the termination end of the first contact area. A conductive path is formed between electrode sensing end one and electrode sensing end two through the blood sample. An 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. At the same time, it commands the electrical module to stop applying a weak electrical signal between the two electrical contacts corresponding to electrode contact one and electrode contact two, and between the two electrical contacts corresponding to electrode contact two and electrode contact six. The timer starts its second countdown, waiting for the blood sample to be tested to react with the magnetic particles and C-reactive protein antibody in area 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. 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 to drive the micro-blood sample containing the aforementioned complex from region two into region three and contact the termination end of region three. Simultaneously, the main control board 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 then formed between electrode sensing terminals two and six through the blood sample containing the aforementioned complex. The 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 the weak electrical signal between the two electrical contacts corresponding to electrode contacts six and five. The main control board then commands the magnetic shield in the magnetic module to move away from the magnet. Between the microfluidic channels, region three is exposed to the magnetic field generated by the magnet. 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 restore the compression pump body to drive the waste liquid that has lost the aforementioned complex from region three into the liquid 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 Zone 1 and Zone 2 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 7

[0119] The present invention is implemented on a microfluidic sample detection system composed of a further form of the microfluidic detection device and a microfluidic detection equipment.

[0120] In this case, Zone 1 is the pretreatment zone, Zones 2 and 3 are the reaction zones, and Zone 4 is the detection zone. For example, using total vitamin D (total 25-hydroxyvitamin D) as the target, Zone 1 contains a pre-set reagent of VD dissociation solution; Zone 2 contains a pre-set reagent of an antibody that binds to the first epitope via magnetic particles (i.e., a first epitope antibody labeled with magnetic particles); Zone 3 contains a pre-set reagent of a second epitope antibody that can be labeled with fluorescein or fluorescent particles; and Zone 4 contains no pre-set reagent. Electrode sensing terminal 5 is located at the beginning of Zone 4, and electrode sensing terminal 7 is located at the end of Zone 4.

[0121] Turn on the power to the testing equipment, place the microfluidic testing device into the load position of the microfluidic testing equipment, and pair the electrical contacts in the testing equipment 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 testing 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 electrode electrical connection signal by manually clicking a button on the testing equipment.

[0122] Upon receiving the signal indicating the electrical connection of the electrodes, the main control board commands the actuator to open valve one and close valve two. With the fluid inlet exposed to the environment, the main control board commands the electrical module to apply a weak electrical signal to the two electrical contacts corresponding to electrodes one and two. The timer begins its first countdown, and simultaneously, the blood sample to be tested is added to the fluid inlet and enters the microfluidic channel. The front end of the blood sample contacts the locking body, stopping the sample from entering. After the first countdown of the timer ends, the main control board confirms successful sample addition and commands the actuator to close valve one, open valve two, and compress the pump to drive the blood sample from valve two to the locking body into region one.

[0123] The blood sample is in contact with the termination point of the first contact area. 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. 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 second countdown of the timer begins, waiting for the blood sample to be fully incubated and dissociated by the VD dissociation solution in area one. In order to fully release the bound VD in the blood sample, the main control board can also command the temperature module to heat area one before the second countdown of the timer begins. Dissociating VD from the blood sample is a specific treatment of bound VD. Only by specifically dissociating it from the blood can subsequent binding reactions be achieved to determine the total VD content in the blood. When the second countdown of the timer ends, the main control board commands the actuator to continue compressing the pump body to drive the blood sample containing the dissociated VD from region one to region two, and commands the electrical module to apply a weak electrical signal between the two electrical contacts corresponding to electrode contact two and electrode contact six. Then, the blood sample containing the dissociated VD contacts the end of region two, that is, between electrode sensing end two and electrode sensing end six, forming a conductive path through the blood sample containing the dissociated VD. The electrical signal can be detected through electrode contact two and electrode contact 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 contact two and electrode contact six. The third countdown of the timer begins, waiting for the blood sample containing the dissociated VD to react with the first epitope antibody labeled by magnetic particles. The dissociated 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. A conductive path is formed between electrode sensing terminals six and five 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 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 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 to form 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 electrical module to apply a weak electrical signal between the two electrical contacts corresponding to electrode contacts five and seven, and commands the actuator to resume the compression pump to drive the micro-blood sample containing the aforementioned complex from region three into region four and contact the termination end of region four. A conductive path is formed between electrode sensing terminals five and seven through the blood sample containing the aforementioned complex, and the electrical signal can be detected through electrode contacts five and seven. 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 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.

[0124] 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.

[0125] 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.

[0126] Example 8

[0127] The present invention is implemented on a microfluidic sample detection system composed of a microfluidic detection device and a microfluidic detection equipment.

[0128] The difference from other embodiments lies in that a pre-prepared liquid reagent is confined within a microfluidic channel within the electrode frame area. When a microfluidic sample flows into the electrode frame area, it reacts with the pre-prepared liquid reagent therein. Furthermore, the pre-prepared liquid reagent is confined within a microfluidic channel within the electrode frame area. When a microfluidic sample flows into the electrode frame area, it reacts with the pre-prepared liquid reagent therein, generating a signal change between the frame electrode and electrode two, which is then transmitted to the microfluidic detection instrument through the frame contact and electrode contact two.

[0129] Example 9

[0130] The present invention is implemented on a microfluidic sample detection system composed of microfluidic detection device 8 and microfluidic detection equipment.

[0131] The difference from other embodiments is that, before the microfluidic detection device is loaded onto the microfluidic detection equipment, after the microfluidic detection equipment is powered on, the main control board commands the electrical detection unit to continuously apply a weak electrical signal between the electrical contacts corresponding to electrode one and electrode eight. After the microfluidic detection device is loaded onto the microfluidic detection equipment, the load point does not generate a signal indicating that it is loaded. The microfluidic sample is added to the microfluidic channel, and the front end of the microfluidic sample contacts electrode sensing end one. A conductive path is formed between electrode sensing end eight and electrode sensing end one through the microfluidic sample to be tested. An electrical signal can be detected through electrode contact eight and electrode contact one, that is, a signal indicating that the microfluidic detection device is loaded is generated. Then, the main control board commands the electrical module to stop applying the weak signal between the two electrical contacts corresponding to electrode contact eight and electrode contact one.

[0132] Alternatively, the difference from other embodiments is that, while the main control board commands the actuator to open valve body one and close valve body two, it commands the electrical module to apply a weak electrical signal to electrode one and electrode eight through contacts. When the front end 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.

[0133] Example 10

[0134] The present invention is implemented on a microfluidic sample detection system composed of a combined microfluidic detection device and a microfluidic detection equipment.

[0135] An apparatus for simultaneously detecting glucose, β-hydroxybutyrate, and hemoglobin in blood, wherein apparatus unit 101 is used to detect glucose, apparatus unit 2 102 is used to detect β-hydroxybutyrate, and apparatus units 3 104 and 4 105 are used to detect hemoglobin.

[0136] 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.

[0137] 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.

[0138] 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.

[0139] Example 11

[0140] The present invention is implemented on a system consisting of one of the microfluidic detection devices and the microfluidic detection equipment. The microfluidic sample detection method of the present invention is used to separate red blood cells and plasma in blood samples and to extract plasma. Red blood cells are used as the target, and the reagent pre-placed in the reaction zone is an antibody that binds to magnetic particles and is designed to fight red blood cells.

[0141] Step 1: 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.

[0142] Step 2: Add the blood sample to the fluid inlet and into 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 front end of the blood sample contacts the lock and stops flowing.

[0143] Step 3: After the first countdown of the timer ends, 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 second valve body to the lock body into the reaction zone. The second countdown of the timer then begins. During the process of the blood sample 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 entering.

[0144] Step 4: When the front end of the blood sample contacts the end of the reaction zone, the second countdown of the timer ends, the main control board commands the actuator to stop the compression pump, and the third countdown of the timer begins.

[0145] Step 5: The blood sample reacts with the antibodies that bind to the magnetic particles in the reaction zone, forming a magnetic particle-anti-erythrocyte antibody-erythrocyte complex.

[0146] Step 6: When the timer finishes its third countdown, 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, thus exposing the reaction zone to the magnetic field generated by the magnet.

[0147] Step 7: Red blood cells bound to antibodies are adsorbed onto the walls of the microfluidic channels in the reaction zone by the magnetic field. The main control board commands the actuator to continue compressing the pump to drive the plasma that has lost red blood cells out of the reaction zone and into the collection zone, and out of the fluid outlet for subsequent use.

[0148] The order of operations between the above steps is not strictly sequential. For example, the operation at the end of the first step and the operation at the beginning of the second step can be performed simultaneously.

[0149] In a further embodiment, one of the aforementioned microfluidic detection devices is replaced with a second microfluidic detection device, the difference being:

[0150] In the first step, when the timer starts its first countdown, the main control board also commands the electrical module to apply a weak current to the two electrical contacts corresponding to electrode one and electrode two.

[0151] In steps three and four, the timer no longer starts its second countdown. The front end of the blood sample contacts electrode sensing terminal two, and 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. The main control board determines that the front end of the blood sample has contacted the termination end of the reaction zone and commands the actuator to stop compressing the pump body. At the same time, it commands the electrical module to stop applying a weak current between the two electrical contacts corresponding to electrode contacts one and two.

[0152] 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.

[0153] 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.

[0154] 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.

[0155] 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.

[0156] 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 method for detecting microfluidic samples, characterized in that, The method is implemented on a microfluidic sample detection system composed of a microfluidic detection device and a microfluidic detection equipment. The microfluidic detection device includes a fluid inlet, a fluid outlet, a microfluidic channel, a liquid collection zone, a reaction zone, a first pump body, a first valve body, a second valve body, and a first lock body; the reaction zone is located in the microfluidic channel between the fluid inlet and the fluid outlet, and the liquid collection zone is located downstream of the reaction zone; the first pump body is located upstream of the reaction zone and is used to passively drive the microfluidic sample to flow in the microfluidic channel; The first valve body is located downstream of the fluid inlet and is used to control the opening and closing between the fluid inlet and the microfluidic channel; the second valve body is located downstream of the first valve body and between the first pump body and the microfluidic channel, and is used to control the opening and closing between the first pump body and the microfluidic channel; the first valve body and the second valve body have opposite working states; the first lock body is located downstream of the second valve body and upstream of the reaction zone termination end; The microfluidic detection device also includes a conductive system composed of electrodes. The conductive system includes at least two electrodes, namely a first electrode and a second electrode. The first electrode includes a first electrode contact and a first electrode sensing end. The second electrode includes a second electrode contact and a second electrode sensing end. The first electrode sensing end and the second electrode sensing end are located at opposite ends of the reaction zone. The microfluidic detection device includes a main control board, an actuator, a timer, an optical module, and an electrical module. 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 detection method is for non-disease diagnosis purposes and includes: Turn on the microfluidic detection device and place the microfluidic detection device in the load position; the microfluidic detection device is signal-connected to the microfluidic detection device. The actuator keeps the first valve body open and the second valve body closed. The microfluidic sample enters the microfluidic detection device through the fluid inlet, and some gas is discharged through the fluid outlet until the front end of the microfluidic sample contacts the first lock body and stops flowing. Once the main control board confirms successful sample addition, the actuator closes the first valve, opens the second valve, and compresses the first pump to drive the microfluidic sample into the reaction zone. When the front end of the microfluidic sample contacts the end of the reaction zone, the actuator stops compressing the first pump body. After the countdown in the timer ends, the actuator resumes compressing the first pump body, driving the microfluidic sample after the reaction to continue flowing away from the reaction zone and enter the liquid collection zone. The optical detection unit in the optical module or the electrical detection unit in the electrical module completes signal detection of the reaction result. The volume of the microfluidic sample that propels forward after entering the microfluidic channel is proportional to the volume of gas discharged from the fluid outlet at the same time.

2. The microfluidic sample detection method according to claim 1, characterized in that, The optical detection unit determines the travel area of ​​the microfluidic sample by continuously detecting the light reflection state.

3. The microfluidic sample detection method according to claim 1, characterized in that, The travel area of ​​the microfluidic sample is determined by the pressure value applied to the first pump body by the actuator.

4. The microfluidic sample detection method according to claim 1, characterized in that, Turn on the microfluidic detection device and place the microfluidic detection device in the load position. The first electrode contact and the second electrode contact are respectively paired with the electrical contacts, and the electrodes of the microfluidic detection device and the microfluidic detection equipment are electrically connected.

5. The microfluidic sample detection method according to claim 4, characterized in that, The electrical module determines the travel area of ​​the microfluidic sample through the first electrode sensing terminal and the second electrode sensing terminal.

6. The microfluidic sample detection method according to claim 4, characterized in that, After the countdown ends, the electrical module receives the reaction result signal through the first electrode sensing end and the second electrode sensing end, and transmits it to the microfluidic detection device through the first electrode contact, the second electrode contact, and the electrical contact.

7. The microfluidic sample detection method according to claim 1, characterized in that, A first region and a second region are formed in the microfluidic channel, the second region being the downstream region, and the first region and / or the second region being the reaction zone. The actuator compresses the first pump body to drive the microfluidic sample to first enter the first region and then the second region. The countdown occurs before the microfluidic sample enters the second region.

8. The microfluidic sample detection method according to claim 7, characterized in that, It also includes a third electrode, which includes a third electrode contact and a third electrode sensing end; the first electrode contact, the second electrode contact and the third electrode contact are respectively paired with electrical contacts to realize the electrical connection of the electrodes of the microfluidic detection device and the microfluidic detection equipment.

9. A microfluidic sample detection method according to claim 8, characterized in that, The electrical module determines the travel region of the microfluidic sample through at least two of the first electrode sensing terminal, the second electrode sensing terminal, and the third electrode sensing terminal.

10. A microfluidic sample detection method according to claim 8, characterized in that, The countdown also occurs after the microfluidic sample enters the second region. After the countdown ends, the electrical module receives the reaction result signal through the second electrode sensing end and the third electrode sensing end, and transmits it to the microfluidic detection device through the second electrode contact, the third electrode contact and the electrical contact.

11. The microfluidic sample detection method according to claim 1, characterized in that, A first region, a second region, and a third region are formed in the microfluidic channel, with the third region being the most downstream region. The first, second, and / or third regions are reaction zones. The actuator compresses the first pump body to drive the microfluidic sample into the first, second, and third regions sequentially. The countdown occurs before the microfluidic sample enters the third region.

12. The microfluidic sample detection method according to claim 11, characterized in that, It also includes a third electrode and a fourth electrode. The third electrode includes a third electrode contact and a third electrode sensing end, and the fourth electrode includes a fourth electrode contact and a fourth electrode sensing end. The first electrode contact, the second electrode contact, the third electrode contact, and the fourth electrode contact are respectively paired with electrical contacts to realize the electrical connection of the electrodes of the microfluidic detection device and the microfluidic detection equipment.

13. The microfluidic sample detection method according to claim 12, characterized in that, The electrical module determines the travel region of the microfluidic sample through at least two of the first electrode sensing terminal, the second electrode sensing terminal, the third electrode sensing terminal, and the fourth electrode sensing terminal.

14. The microfluidic sample detection method according to claim 12, characterized in that, The countdown also occurs after the microfluidic sample enters the third region. After the countdown ends, the electrical module receives the reaction result signal through the third electrode sensing end and the fourth electrode sensing end, and transmits it to the microfluidic detection device through the third electrode contact, the fourth electrode contact, and the electrical contact.

15. A microfluidic sample detection method according to claim 1, characterized in that, A first region, a second region, a third region, and a fourth region are formed in the microfluidic channel, with the fourth region being the most downstream region. The first, second, third, and / or fourth regions are reaction zones. The actuator compresses the first pump body to drive the microfluidic sample into the first, second, third, and fourth regions sequentially. The countdown occurs before the microfluidic sample enters the fourth region.

16. A microfluidic sample detection method according to claim 15, characterized in that, It also includes a third electrode, a fourth electrode, and a fifth electrode. The third electrode includes a third electrode contact and a third electrode sensing end. The fourth electrode includes a fourth electrode contact and a fourth electrode sensing end. The fifth electrode includes a fifth electrode contact and a fifth electrode sensing end. The first electrode contact, the second electrode contact, the third electrode contact, the fourth electrode contact, and the fifth electrode contact are respectively paired with electrical contacts to realize the electrical connection of the electrodes of the microfluidic detection device and the microfluidic detection equipment.

17. A microfluidic sample detection method according to claim 16, characterized in that, The electrical module determines the travel region of the microfluidic sample through at least two of the first electrode sensing terminal, the second electrode sensing terminal, the third electrode sensing terminal, the fourth electrode sensing terminal, and the fifth electrode sensing terminal.

18. A microfluidic sample detection method according to claim 16, characterized in that, The countdown also occurs after the microfluidic sample enters the fourth region. After the countdown ends, the electrical module receives the reaction result signal through the fourth electrode sensing end and the fifth electrode sensing end, and transmits it to the microfluidic detection device through the fourth electrode contact, the fifth electrode contact, and the electrical contact.

19. A microfluidic sample detection method according to any one of claims 7-18, characterized in that, The optical module obtains the reaction result signal by detecting the downstream region.

20. A microfluidic sample detection method according to any one of claims 1-18, characterized in that, The reaction zone includes a target antibody labeled with magnetic particles, which specifically binds to the target in the microfluidic sample.

21. A microfluidic sample detection method according to any one of claims 1-18, characterized in that, The reaction zone includes fluorescently labeled antibodies that specifically bind to targets in the microfluidic sample.

22. A microfluidic sample detection method according to any one of claims 7-18, characterized in that: Microfluidic detection equipment also includes a magnetic module; The reaction zone includes a first reaction zone and a second reaction zone, in which a magnetically labeled target antibody and a fluorescently labeled target antibody are pre-placed, respectively, in the first and second reaction zones. The actuator compresses the first pump body to drive the microfluidic sample into the first reaction zone and then into the second reaction zone, where the target analyte, the first antibody, and the second antibody together form a bi-antibody sandwich complex. Before the actuator compresses the first pump body and drives the reacted microfluidic sample to continue flowing, the magnetic module generates a magnetic field to adsorb the dual-antibody sandwich composite onto the wall of the microfluidic channel.

Images