Microfluid sample detection method

Abstract

The invention provides a microfluid sample detection method, which comprises the following steps of: firstly, opening microfluid detection equipment, placing the microfluid detection equipment into a load position, and connecting the microfluid detection equipment with the microfluid detection device through a signal; and secondly, the treatment, reaction and detection of the microfluid sample are completed by controlling the first pump body, the first valve body, the second valve body, the third valve body and the third valve body, the actuator executes various first pump body actions to drive the microfluid sample to flow from the fluid inlet to the fluid outlet in the microfluid channel, and the treatment, reaction and detection are carried out in the process. And finally, after the countdown in the timer is finished, the actuator compresses the first pump body, and the reacted microfluid sample is driven to continuously flow and enter the liquid collecting area away from the reaction area. According to the invention, single detection and all-in-one combined detection can be carried out, and even electrochemical detection and photochemical detection methods can be combined for detection.

Description

Technical Field

[0001] The present invention relates to the field of medical devices, and in particular to a microfluid sample detection method. Background Art

[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 the quality of people's lives and increased the efficiency of their study and work.

[0003] Microfluidic chips are a type of science and technology that is mainly characterized by the manipulation of fluids in micrometer-scale space. They have the ability to miniaturize the basic functions of biological and chemical laboratories onto a chip of a few square centimeters, so they are also called chip laboratories. At present, the mainstream form of microfluidic chips is mostly formed by a network of microchannels, with controllable fluids running through the entire system to achieve various functions of conventional chemical or biological laboratories. The basic characteristics and greatest advantage of microfluidic chips are the flexible combination and large-scale integration of multiple unit technologies on a tiny controllable platform. It is very suitable for the design of products in the bedside point-of-care diagnostic industry (POCT). However, the materials commonly used to make microfluidic chips are glass and organic polymers such as polymethyl methacrylate (PMMA), polydimethylsiloxane (PDMS), polycarbonate (PC) and hydrogel. Its processing and production environment includes air temperature, air humidity, and particle density in the air and various media used in the preparation process. The higher environmental requirements for chip production generally need to be achieved in a clean room. Clean room technology is closely related to the success or failure of the microfluidic chip manufacturing process. Depending on the processing and manufacturing method, the clean room standard must reach Class 10,000, Class 1,000 or even Class 100. For example, the manufacturing technology used in the production of polymer microfluidic chips mainly includes hot pressing, molding, injection molding, laser ablation, LIGA, etc. However, the equipment used in the prior art is expensive and the steps are complicated. Not only does it have strict requirements on the environmental quality during the process and high operating costs, but the output per batch is low, so the cost of manufactured products is generally high. There is still a certain gap between the production capacity demand and cost requirements of bedside instant diagnosis (POCT) products, which limits the popularity of microfluidic technology in the POCT industry.

[0004] To address the above-mentioned problems, no effective solution has been proposed yet. Summary of the invention

[0005] In order to solve the above problems, the present invention provides a microfluid sample detection method, comprising: opening a microfluid detection device and placing the microfluid detection device in a load position, the microfluid detection device is connected to the microfluid detection device by signal; an actuator opens a first valve body and a third valve body, closes a second valve body and a fourth valve body, and compresses a first pump body to drive at least part of the gas in the first pump body to be discharged from a fluid inlet through the third valve body, or opens the first valve body, the second valve body and the third valve body, closes the fourth valve body, compresses the first pump body to drive at least part of the gas in the first pump body to be discharged from the fluid inlet through the second valve body and the third valve body, and then closes the second valve body. body; in response to the signal that the microfluid sample contacts the fluid inlet, the main control board commands the release of the first pump body to drive the microfluid sample into the microfluid channel. After the main control board determines that the sample is added successfully, it commands to stop releasing the first pump body and close the first valve body; the actuator executes the action of the first pump body to drive the microfluid sample into the reaction area, and closes the third valve body, opens the second valve body, and opens the fourth valve body before or after executing the action; the actuator stops the action of the first pump body, waits for the countdown in the timer to end, and compresses the first pump body to drive the microfluid sample after the reaction to continue to flow downstream into the liquid collection area, and performs reaction result signal detection on the reaction area. At least part of the gas is discharged from the fluid inlet, which means that part or all of the gas is discharged from the fluid inlet, which depends on the design of the structure of the microfluid detection device. If the microfluid channel between the second valve body and the third valve body of the microfluid detection device is very long to complete the process of processing, reacting and detecting the microfluid sample, the gas in the first pump body can be completely discharged. If the microfluid channel between the second valve body and the third valve body of the microfluid detection device is very short, the gas in the first pump body can be only partially discharged.

[0006] In some embodiments, after the main control board determines that the sample loading is successful and commands to stop releasing the first pump body and close the first valve body, the actuator executes the action of releasing the first pump body to drive the microfluidic sample into the reaction area, and then the actuator closes the third valve body and opens the second valve body and the fourth valve body.

[0007] In other embodiments, after the actuator releases the first pump body, it opens the second valve body before the microfluid sample fills the reaction area, and compresses the first pump body to drive the microfluid sample to fill the reaction area.

[0008] In a preferred embodiment, after the main control board determines that the sample loading is successful, it commands to stop releasing the first pump body, close the first valve body and the third valve body, and open the second valve body and the fourth valve body. Then, the actuator performs the action of compressing the first pump body to drive the microfluidic sample into the reaction area.

[0009] In another embodiment, after the microfluidic detection device is connected to the microfluidic detection equipment signal, the actuator opens the first valve body, the second valve body and the third valve body, closes the fourth valve body, closes the second valve body before driving the microfluidic sample into the reaction area, and the actuator releases the first pump body to drive the microfluidic sample into the reaction area.

[0010] Furthermore, after the actuator releases the first pump body, the second valve body is opened before the microfluid sample fills the reaction area, and the first pump body is compressed to drive the microfluid sample to fill the reaction area. In another embodiment, the actuator compresses the first pump body to drive the microfluid sample from the first area into the second area, and the countdown occurs before the microfluid sample enters the second area.

[0011] Specifically, the light detection unit determines the travel area of ​​the microfluidic sample by continuously detecting the light reflection state. The travel area includes the area where the sample is successfully added, the area where the microfluidic sample is processed, the area where the microfluidic sample reacts, and the area where the microfluidic sample is detected.

[0012] Specifically, the travel area of ​​the microfluid sample is determined by the pressure value applied by the actuator to the first pump body. The travel area includes the area where the sample is successfully added, the area where the microfluid sample is processed, the area where the microfluid sample is reacted, and the area where the microfluid sample is detected.

[0013] In another embodiment, the microfluidic detection device is opened and the microfluidic detection device is placed in the load position, the first electrode contact and the second electrode contact are respectively paired with the electrical contact pins, and the microfluidic detection device and the microfluidic detection device electrodes are electrically connected.

[0014] Preferably, the electrical module determines the travel area of ​​the microfluidic sample through the first electrode sensing end and the second electrode sensing end. The travel area includes the area where the sample is successfully added, the area where the microfluidic sample is processed, the area where the microfluidic sample reacts, and the area where the microfluidic sample is detected.

[0015] Furthermore, 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 pin.

[0016] Or further, the actuator performs an action of compressing the first pump body to drive the microfluid sample from the first area into the second area, and the countdown occurs before the microfluid sample enters the second area.

[0017] In another embodiment, the microfluidic detection device is opened and the microfluidic detection device is placed in the load position, the first electrode contact, the second electrode contact and the third electrode contact are respectively paired with the electrical contact pins, and the microfluidic detection device and the microfluidic detection device electrodes are electrically connected.

[0018] Furthermore, the electrical module determines the travel area of ​​the microfluidic sample through at least two of the first electrode sensing end, the second electrode sensing end, and the third electrode sensing end. The travel area includes a region where the sample is successfully added, a region where the microfluidic sample is processed, a region where the microfluidic sample reacts, and a region where the microfluidic sample is detected.

[0019] Or further, the countdown also occurs after the microfluidic sample enters the second area. 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 and the third electrode contact, as well as the electrical contact pin.

[0020] In another embodiment, the microfluidic sample enters the first area, the second area, and the third area in sequence, and the countdown occurs before the microfluidic sample enters the third area.

[0021] Furthermore, it is characterized in that the first electrode contact, the second electrode contact, the third electrode contact and the fourth electrode contact are respectively paired with electrical contact pins to achieve electrical connection between the microfluidic detection device and the electrodes of the microfluidic detection equipment.

[0022] Or further, the electrical module determines the travel area of ​​the microfluidic sample through at least two of the first electrode sensing end, the second electrode sensing end, the third electrode sensing end and the fourth electrode sensing end. The travel area includes the area where the sample is successfully added, the area where the microfluidic sample is processed, the area where the microfluidic sample is reacted, and the area where the microfluidic sample is detected.

[0023] Or further, the countdown also occurs after the microfluidic sample enters the third area. 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 and the fourth electrode contact, as well as the electrical contact pin.

[0024] In another embodiment, the microfluidic sample enters the first area, the second area, the third area, and the fourth area in sequence, and the countdown occurs before the microfluidic sample enters the fourth area.

[0025] 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 contact pins to achieve electrical connection between the microfluidic detection device and the electrodes of the microfluidic detection equipment.

[0026] Or further, the electrical module determines the travel area of ​​the microfluidic sample through at least two of the first electrode sensing end, the second electrode sensing end, the third electrode sensing end, the fourth electrode sensing end and the fifth electrode sensing end. The travel area includes the area where the sample is successfully added, the area where the microfluidic sample is processed, the area where the microfluidic sample is reacted, and the area where the microfluidic sample is detected.

[0027] Or further, the countdown also occurs after the microfluidic sample enters the fourth area. 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 and the fifth electrode contact, as well as the electrical contact pin.

[0028] In another embodiment, the optical module obtains a reaction result signal by detecting the most downstream area. In another embodiment, the reaction area includes a target antibody labeled with magnetic particles, and the antibody can specifically bind to the target in the microfluidic sample.

[0029] In other embodiments, the reaction zone includes antibodies labeled with fluorescent particles, and the antibodies specifically bind to targets in the microfluidic sample.

[0030] In another embodiment, the reaction area includes a first reaction area and a second reaction area, and the first target antibody labeled with magnetic particles and the second target antibody labeled with fluorescent particles are pre-placed in the first reaction area and the second reaction area respectively;

[0031] The actuator performs a first pump action to drive the microfluidic sample to first enter the first reaction zone and then enter the second reaction zone, and the target object, the first antibody and the second antibody together form a double 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 double antibody sandwich complex on the wall of the microfluidic channel.

[0033] The beneficial effects of the present invention include: 1. It can be applied to in vitro diagnostic products of various methodologies, and can perform single-item detection, all-in-one combination detection, and can even combine electrochemical detection and photochemical detection methods into the same detection process, and simultaneously detect microfluidic samples entering from a fluid inlet; 2. The microfluidic sample after reaction enters the liquid collection area at the most downstream, and there will be no problem of contaminating the detection equipment; 3. In the process of microfluidic sample detection, by designing a variety of execution actions of the pump body, more control methods can be provided for the flow of microfluidic samples, and the scope of application can be further broadened. BRIEF DESCRIPTION OF THE DRAWINGS

[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 implementation mode of the microfluidic detection device of the present invention.

[0037] Figure 1-4 A fourth implementation mode of the microfluidic detection device of the present invention.

[0038] Figure 2 A fifth implementation of the microfluidic detection device of the present invention.

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

[0040] Figure 3-2 Embodiment 7 of the microfluidic detection device of the present invention.

[0041] Figure 4-1 Embodiment 8 of the microfluidic detection device of the present invention.

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

[0043] Figure 5 A combined implementation mode of the microfluidic detection device of the present invention.

[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] Fig. 9 Example 6 Results of a study on the accuracy of the product of the present invention.

[0048] Fig.10 Example 6A Results of a study on the accuracy of conventional products.

[0049] Fig.11 Example 6B Results of a study on the accuracy of conventional products. DETAILED DESCRIPTION

[0050] In order to enable those skilled in the art to better understand the solutions of the present invention, the following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

[0051] Microfluidic detection device

[0052] like Figure 1-1 As shown, one of the microfluidic detection devices implementing the present invention includes a fluid inlet 11, a fluid outlet 12, a microfluidic channel 13, a liquid collection area 14, a reaction area 131, a pump body 41, and a valve body.

[0053] The fluid inlet 11 is used to add the microfluid sample to be tested, the fluid outlet 12 is connected to the environment, the microfluid channel 13 is between the fluid inlet and the fluid outlet, the microfluid sample to be tested enters from the fluid inlet and flows in the microfluid channel, and the volume of the microfluid sample pushed forward in the microfluid channel is greater than the volume of the gas discharged from the fluid outlet. The liquid receiving area 14 is downstream of the microfluid channel and also downstream of the reaction area, and is used to receive the microfluid sample after the reaction. The fluid outlet is located at the outlet of the liquid receiving area and is connected to the environment. The environment refers to the external environment of the microfluid detection device of the present invention, such as air and vacuum, which can be the environment set in the system of the present invention, or it can be the same as the environment outside the system of the present invention. The fluid outlet allows gas or liquid to flow out. In most embodiments, the fluid outlet is a gas outlet, which only allows gas to flow out, and liquid cannot flow out; in a small number of embodiments, the fluid outlet is an outlet for gas and liquid, which allows both gas and liquid to flow out.

[0054] The reaction zone 131 is in the microfluidic channel between the fluid inlet and the fluid outlet. In other words, the reaction zone overlaps with part of the microfluidic channel. The microfluidic sample to be tested is designed to react with the fixed preset reagent in the reaction zone during the flow in the microfluidic channel. At this time, the reaction zone is the most downstream area.

[0055] The pump body 41 is used to passively drive the microfluid sample to flow in the microfluid channel. In other words, by controlling the pressure applied to the pump body, a driving force can be provided for the flow of the microfluid sample, and the driving force drives the microfluid sample to be tested to enter the microfluid channel from the fluid inlet, and the driving force can further drive the microfluid sample to be tested to continue to flow downstream. The pressure value applied to the pump body can be preset and associated with the travel position of the microfluid sample in the microfluid channel after entering the microfluid channel. That is to say, when the pressure value applied to the pump body is preset to a certain value, the corresponding microfluid sample is at a certain position in the microfluid channel. This design can be used to confirm that the microfluid sample is successfully added and to determine that the microfluid sample has entered a certain area. The valve body includes valve body one 31, valve body two 32, valve body three 33 and valve body four 33. Valve body one 31 is arranged downstream of the fluid inlet, and is used to control the on-off between the fluid inlet and the microfluid channel. It is in an open state before the microfluid sample enters the microfluid channel. The second valve body 322 and the third valve body 33 are arranged downstream of the first valve body, and are both located between the pump body and the microfluidic channel, and are used to control the connection or disconnection between the pump body and the microfluidic channel. When the second valve body and / or the third valve body are opened, the pump body and the microfluidic channel fluid are connected. In other words, the second valve body and the third valve body control the pump body and determine whether the pump body provides driving force for the microfluidic sample. The second valve body and the third valve body can be in an open state for a period of time at the same time, but cannot be in an open state at the same time all the time. The third valve body is located downstream of the second valve body, the second valve body is upstream of the reaction zone, and the third valve body can be upstream of the reaction zone or downstream of the reaction zone, that is, the reaction zone can be located in the microfluidic channel between the second valve body and the third valve body, or in the microfluidic channel downstream of the third valve body. The first valve body and the third valve body jointly determine the volume of the microfluidic sample that finally participates in the reaction, or the second valve body and the third valve body jointly determine the volume of the microfluidic sample that finally participates in the reaction. The fourth valve body is arranged at the fluid outlet, and is used to control the connection and disconnection between the liquid receiving area and the external environment. The working states of the third valve body and the fourth valve body are opposite, that is, the fourth valve body is closed when the third valve body is opened, and the third valve body is closed when the fourth valve body is opened. When the third valve body is opened and the fourth valve body is closed, the pump body and the microfluidic channel are connected, and the pump body can passively drive the microfluidic sample to flow toward the fluid outlet. When the third valve body is opened, the second valve body can be opened at the same time to drive the microfluidic sample to flow toward the fluid outlet, but before the front end of the microfluidic sample approaches the second valve body, the second valve body must be closed to prevent the microfluidic sample from entering the pump body through the second valve body. Similarly, before the front end of the microfluidic sample approaches the third valve body, the third valve body must be closed to prevent the microfluidic sample from entering the pump body through the third valve body.

[0056] In some implementations, the valve body is composed of a plastic substrate and a flexible diaphragm, the plastic substrate includes an upper surface and a lower surface, a groove is formed on the upper surface by laser, etching or other methods, the flexible diaphragm 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 by an adhesive material, the upper surface of the plastic substrate and the lower surface of the flexible diaphragm are in contact and partially joined, a cavity is formed between the upper surface of the plastic substrate and the lower surface of the flexible diaphragm, the cavity is connected when the flexible diaphragm is in a relaxed state, that is, the valve body is in an open state, and the cavity is not connected under pressure when the flexible diaphragm is in a closed state.

[0057] In other embodiments, the valve body is composed of a plastic substrate, a double-sided adhesive and a flexible diaphragm, the plastic substrate includes an upper surface and a lower surface, the double-sided adhesive includes a hollow, 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, and a cavity is formed 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 connected, that is, the valve body is in an open state. When the flexible diaphragm is under pressure, the cavity is not connected, that is, the valve body is in a closed state.

[0058] like Figure 1-2 As shown, the second microfluidic detection device of the present invention is implemented, based on the first microfluidic detection device, further comprising a conductive system composed of electrodes. The conductive system comprises two electrodes, electrode 1 51 and electrode 2 52, wherein electrode 1 further comprises electrode contact 1 511 and electrode sensing end 1 512, and electrode 2 further comprises electrode contact 2 521 and electrode sensing end 2 522, and electrode sensing end 1 512 and electrode sensing end 2 522 are respectively located at the two ends of the reaction zone 131, that is, electrode sensing end 1 is located at the starting end of the reaction zone, and electrode sensing end 2 is located at the ending end of the reaction zone, as shown in FIG. Figure 1-2 As shown in (a), the electrode sensing end 1 and the electrode sensing end 2 are both located downstream of the valve body 2 and between the valve body 2 and the valve body 3. At this time, the reaction zone is between the valve body 2 and the valve body 3; Figure 1-2 As shown in (b), the electrode sensing end 1 and the electrode sensing end 2 are both located downstream of the valve body 3, and the reaction zone is downstream of the valve body 3; Figure 1-2 As shown in (c), the electrode sensing end 1 is downstream of the valve body 2 and upstream of the valve body 3, and the electrode sensing end 2 is downstream of the valve body 3. At this time, the valve body 3 is within the range of the reaction zone.

[0059] like Figure 1-3As shown, the third microfluidic detection device for implementing the present invention is based on one of the microfluidic detection devices or the second microfluidic detection device or other subsequent microfluidic detection devices, and also includes a microfluidic channel branch 132, a branch outlet 121, and an electrode three 53, an electrode four 54, an electrode contact three 531, an electrode sensing end three 532, an electrode contact four 541, and an electrode sensing end four 542, wherein the branch outlet is a gas outlet that only allows gas to flow out.

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

[0061] like Figure 2 As shown, the fifth microfluidic detection device for implementing the present invention is based on the second microfluidic detection device, and the conductive system also includes an electrode five 55, that is, the electrode system includes three electrodes, and the electrode five includes an electrode contact five 551 and an electrode sensing end five 552. Region one 1311 and region two 1312 are formed in the microfluidic channel, and at this time, region two is the most downstream region. Among them, region one is a reaction zone or region two is a reaction zone or region one and region two are both reaction zones. When the pump body and the microfluidic channel are connected through valve body two and / or the third valve body, the pump body can drive the microfluidic sample to enter region one and region two in sequence. 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 also the starting end of region two, 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 the positional relationship between electrode sensing end one and electrode sensing end two relative to valve body two and valve body three and Figure 1-2 Same as in. Figure 1-2 As shown in (a), area 1 is located upstream of valve body 3; Figure 1-2 As shown in (b), area 1 is located downstream of valve body 3; Figure 1-2 As shown in (c), valve body three is within the range of area one.

[0062] like Figure 3-1As shown, the sixth microfluidic detection device of the present invention is implemented. On the basis of the fifth microfluidic detection device, the conductive system also includes an electrode six 56, that is, the electrode system includes four electrodes, and the 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. At this time, region three is the most downstream region. Among them, region one is a reaction area or region two is a reaction area or region three is a reaction area or any two of region one, region two and region three are reaction areas or region one, region two and region three are all reaction areas. When the pump body and the microfluidic channel are connected through the second valve body and / or the third valve body, the pump body can drive the microfluidic sample to enter region one, region two and region three in sequence. Electrode sensing end one is located at the starting end of region one, electrode sensing end two is located at the termination end of region one and is also the starting end of region two, electrode sensing end six is ​​located at the termination end of region two and is also the starting end of region three, and electrode sensing end five is located at the termination end of region three.

[0063] Furthermore, it also includes electrode seven, which includes electrode contact seven and electrode sensing end seven, and forms region one, region two, region three and region four in the microfluidic channel, that is, electrode sensing end five is located at both the terminal end of region three and the starting end of region four, and electrode sensing end seven is located at the terminal end of region four, and at this time, region four is the most downstream region. Among them, region one is a reaction area, region two is a reaction area, region three is a reaction area, or region four is a reaction area, or any two / three of region one, region two, region three and region four are reaction areas, or region one to region four are all reaction areas, and when the pump body and the microfluidic channel are connected through valve body two and / or the third valve body, the pump body can drive the microfluidic sample to enter region one, region two, region three and region four in sequence.

[0064] like Figure 3-2 As shown, the seventh microfluidic detection device of the present invention is implemented, based on the fifth microfluidic detection device, further comprising a frame electrode 501, an electrode frame area 5011, and a frame contact 5012, or only comprising the electrode frame area 5011. The microfluidic detection device is formed by bonding a substrate and a covering film, the frame electrode is printed on the substrate, the electrode frame area is printed on the covering film and protrudes from the covering film, when the substrate and the covering film are bonded together, the electrode frame area and the microfluidic channel in the substrate jointly form an area for the collection of pre-set reagents, and the two sides of the electrode frame area parallel to each other in the horizontal direction can be used as electrode sensing ends of the frame electrode.

[0065] like Figure 4-1As shown, the eighth microfluidic detection device of the present invention is implemented on the basis of any one of the first to seventh microfluidic detection devices or other subsequent microfluidic detection devices, including electrode one 51, electrode contact one 511, electrode sensing end one 512, electrode eight 58, electrode contact eight 581, electrode sensing end eight 582, electrode sensing end eight is located downstream of valve body one and upstream of valve body two, electrode sensing end one is located downstream of electrode sensing end eight, electrode sensing end one, electrode sensing end eight and microfluidic channel are jointly surrounded to form a start zone, the start zone is close to the microfluid inlet, electrode sensing end eight is the starting end of the start zone, and electrode sensing end one is the ending end of the start zone. Or, Figure 4-2 As shown, the electrode sensing terminal 8 may also be located upstream of one of the valve bodies and downstream of the fluid inlet.

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

[0068] Microfluidic testing equipment

[0069] like Figure 6 , Figure 7 , Figure 8 As shown, it is a schematic diagram of the structure of the microfluidic detection device 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 a specific implementation, the optical module and the electrical module include at least one of them; the magnetic module, the temperature module, and the display screen are optional designs, which may be included in the microfluidic detection device or may not be included in the microfluidic detection device.

[0070] The power interface provides power for the detection device and can charge the battery. The battery helps the detection device to work normally without power access, thus realizing the portability of the detection device. After the detection device is turned on, the detection device is taken out and placed in the detection device. When the microfluidic sample detection device is accurately placed on the load position in the detection device, the load position will generate a loaded signal and transmit it to the main control board. The detection device and the detection device complete the signal connection, and the main control board commands the detection device to start running. In another embodiment, the main control board can command the detection device to start running by clicking a button on the detection device.

[0071] At least one reaction waiting time countdown is pre-set in the timer to determine a fixed time period for the reaction of the microfluidic sample in the microfluidic channel. In another embodiment, a countdown for the waiting time for adding the microfluidic sample can also be set in the timer, and the time when the detection device and the detection equipment complete the signal connection is used as the starting point of the waiting time for adding the sample. When the time in the timer passes, it is determined that the microfluidic sample to be tested has been successfully added. When the timer is used to determine that the addition is successful, there is no need to use the above-mentioned light detection unit to determine that the addition is successful. In another embodiment, a countdown for the time when the microfluidic sample reaches a fixed area (such as a reaction area) can also be set in the timer. The countdown starts when the microfluidic sample starts to flow, and when the countdown ends, the microfluidic sample reaches the fixed area.

[0072] The actuator provides pressure to the pump body of the detection device, provides pressure to the pump body after the detection device is accurately placed on the load position of the detection device, compresses the pump body to drive part or all of the gas in the pump body to flow out of the detection device; maintains the pressure provided to the pump body, and releases the pump body to drive the microfluidic sample to be tested to flow and enter the microfluidic channel when the microfluidic sample to be tested contacts the fluid inlet; and provides pressure to the pump body after the microfluidic sample to be tested is successfully added to the microfluidic channel, compresses the pump body to drive the microfluidic sample to flow toward the downstream area of ​​the microfluidic channel, where the downstream area refers to, for example, the processing area, the reaction area, the detection area, etc. In some embodiments, the pressure value applied by the actuator to the pump body can be pre-set and associated with the travel position of the microfluidic sample in the microfluidic channel after entering the microfluidic channel, that is, when the pressure value applied to the pump body is pre-set to a certain value, the corresponding microfluidic sample is at a certain position in the microfluidic channel. This design can be used to confirm that the microfluidic sample is successfully added and to determine that the microfluidic sample has entered a certain area. For example, when the pressure value applied by the actuator reaches a certain value, the microfluidic sample flows to the reaction area. In such an embodiment, the timer may not need to set a countdown for the time when the microfluidic sample reaches the fixed area. The actuator provides pressure to the valve body to close the valve body so that the microfluidic channel is closed, or releases pressure to open the valve body so that the microfluidic channel is connected.

[0073] The optical module includes a light detection unit, which detects the reaction result of the microfluid sample to be tested, and then the main control board processes it and displays the detection result on the display screen. Alternatively, the optical module also includes a photoelectric conversion unit, which converts the photoelectric conversion unit into an electrical signal, which is then processed by the main control board and then displayed on the display screen.

[0074] On the other hand, the optical module is also used to determine whether the microfluid sample to be tested has been successfully added to the detection device. The light detection unit continuously detects the light reflection state of the same position in the microfluid channel at different time points and converts it into an electrical signal by the photoelectric conversion unit. The main control board determines that the microfluid sample to be tested has been successfully added.

[0075] The electrical module includes an electrical connector and an electrical detection unit. The electrical connector provides paired electrical contact pins for the electrode contacts in the microfluid detection device. The number of electrode contacts in the microfluid detection device corresponds to the number of electrical contact pins in the corresponding microfluid detection device. The main control board commands the electrical detection unit to output a continuous weak electrical signal (the output stops after the microfluid sample to be tested is successfully added). It cannot be detected in the case of a circuit break. When the detection device has been accurately placed on the load position of the detection device, the electrode contacts on the detection device are paired with the electrical contact pins on the electrical connector, indicating that the detection device and the detection device have completed the electrical connection of the electrodes. The electrical module determines that the microfluid sample to be tested has been successfully added by detecting the electrical signal between the electrodes. Alternatively, since the time for a microfluid sample of a certain volume to pass through a microfluid channel of a certain size is determined, the sample addition waiting time of the microfluid sample is set in the timer in advance, and the time when the detection device and the detection device complete the signal connection is used as the starting point of the sample addition waiting time timing. When the time in the timer passes, it is determined that the microfluid sample to be tested has been successfully added.

[0076] After the microfluid sample to be tested 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 electrode contact one and electrode contact two through the corresponding electrical contact pins. After processing by the main control board, the detection result is displayed on the display screen.

[0077] The magnetic module includes a magnet and a magnetic shielding body. The magnet is fixed near the downstream area of ​​the microfluidic channel in the detection device, and the magnet is isolated from the downstream area of ​​the microfluidic channel by the magnetic shielding body. When the microfluidic sample to be tested enters the downstream area of ​​the microfluidic channel and the microfluidic sample to be tested needs to be magnetically adsorbed and separated, the main control board commands the magnetic shielding body to move so that the magnetic field generated by the magnet acts on the microfluidic sample to be tested.

[0078] The temperature module includes a heating unit, which is fixed in a downstream area close to 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 for the microfluidic sample to be tested.

[0079] Microfluidic sample extraction system

[0080] The system formed by combining the above-mentioned microfluidic sample detection device and the microfluidic sample detection equipment is applied to the separation and extraction of microfluidic samples.

[0081] Example 1

[0082] The present invention is implemented on a microfluidic sample detection system composed of one of the microfluidic detection devices and a microfluidic detection equipment, and urine sugar is detected using the microfluidic sample detection method of the present invention. Glucose is used as the target, and the reagent pre-installed in the reaction zone is prepared by disodium hydrogen phosphate, citric acid, polyvinyl pyrrolidone, dimethyl maleate, o-tolidine, peroxidase, glucose oxidase, orange G dye, enzyme stabilizer, and enzyme activator.

[0083] Turn on the power of the detection device, take the microfluidic detection device and put it on the load position of the microfluidic detection device, and expose the fluid inlet to the environment. The load position generates a loaded signal and transmits it to the main control board. In another embodiment, the load position does not generate a loaded signal, but transmits the loaded signal to the main control board by manually clicking a button on the detection device.

[0084] The microfluidic detection device is connected to the microfluidic detection equipment signal. The main control board commands the actuator to open valve body one, open valve body three, close (or open) valve body two, close valve body four, compress the pump body to drive the gas in the pump body through valve body three (or, and valve body two), and then discharge it through the fluid inlet. The main control board commands to stop compressing the pump body, keep valve body two closed (or closed), and contact the urine sample to be tested with the fluid inlet. The first countdown of the timer starts. At the same time, the main control board commands to release the pump body to drive the microfluidic sample into the microfluidic channel. During the process of the urine sample to be tested entering the microfluidic channel, part of the gas enters the pump body through valve body three. The volume of gas entering the pump body is proportional to the volume of the urine sample to be tested that enters.

[0085] When the first countdown of the timer ends, the urine sample to be tested stops entering. The main control board receives the countdown end signal to determine that the sample addition has been successful, and instructs the actuator to close valve body one, open valve body two, close valve body three, open valve body four, and compress the pump body to drive the urine sample to be tested between valve body two and valve body three into the reaction zone. The second countdown of the timer begins. During the process of the urine sample to be tested entering the reaction zone, part of the gas is discharged through the fluid outlet, and the volume of the discharged gas is proportional to the volume of the urine sample to be tested traveling downstream.

[0086] The front end of the urine sample to be tested contacts the termination end of the reaction zone, that is, the urine sample has filled the reaction zone, the second countdown of the timer ends, the main control board commands the actuator to stop compressing the pump body, the third countdown of the timer begins, the urine sample to be tested reacts with the preset reagent in the reaction zone, the third countdown of the timer ends, the main control board commands the optical detection unit in the optical detection module to perform optical signal detection of the reaction result on the reaction zone and 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 there is glucose in the urine sample and commands the display screen to display the test result, and at the same time commands the actuator to resume compressing the pump body to drive the waste liquid after the reaction to continue to flow and enter the liquid collection area. When the waste liquid after the reaction enters the liquid collection area, part of the gas is discharged through the fluid outlet, the liquid collection area absorbs the waste liquid after the reaction, and the used microfluidic sample detection device is taken out of the detection equipment and discarded.

[0087] In another embodiment, after the gas in the pump body is discharged, the main control board commands to stop compressing the pump body, opens (or keeps open) the second valve body, and allows the urine sample to be tested to contact the fluid inlet. The first countdown of the timer starts, and at the same time, the main control board commands to release the pump body to drive the urine sample to be tested into the microfluidic channel. During this process, part of the gas enters the pump body through the second valve body and the third valve body. The volume of gas entering the pump body is proportional to the volume of the urine sample to be tested that enters. When the front end of the urine sample to be tested approaches the second valve body, the main control board commands to close the second valve body, but continues to keep the third valve body open.

[0088] In another embodiment, there is no first countdown, but the successful addition of the sample is determined by observation, and a cover body that can generate a successful addition signal is configured for the detection device. When the urine sample to be tested is added to the fluid inlet and the front end of the urine sample to be tested is observed to contact the lock body and stop flowing, the cover body is closed, and the cover body generates a successful addition signal and transmits it to the main control board to determine the successful addition of the sample; in another embodiment, there is no first countdown, and the urine sample is successfully added by the optical module; in another embodiment, there is no second countdown, and the pressure value applied by the actuator on the pump body is used to determine that the urine sample has filled the reaction area. In another embodiment, the first countdown is not required, but the successful addition of the sample is determined by observation, and a cover body that can generate a successful addition signal is configured for the detection device. When the urine sample to be tested is added to the fluid inlet and it is observed that the amount of the urine sample to be tested is sufficient, the cover body is closed, and the cover body generates a successful addition signal and transmits it to the main control board.

[0089] Example 2

[0090] The present invention is implemented on a microfluidic sample detection system composed of a second microfluidic detection device and a microfluidic detection equipment, and blood ketone detection is performed using the microfluidic sample detection method of the present invention. β-hydroxybutyric acid is used as the target, and the reagent pre-installed in the reaction area 131 is a preparation formed by β-hydroxybutyrate dehydrogenase, an electron mediator, a surfactant, and a buffer solution.

[0091] Turn on the power of the detection device, take the microfluidic detection device and put it into the load position of the microfluidic detection device, the electrical contact pins in the detection device are matched with the electrode contact 1 and the electrode contact 2, the load position generates a loaded signal and transmits it to the main control board, indicating that the detection device and the detection device have completed the electrode electrical connection. In another embodiment, the load position does not generate a loaded signal, but manually clicks the button on the detection device to transmit the loaded signal to the main control board, indicating that the detection device and the detection device have completed the electrode electrical connection. The main control board receives the signal of the electrode electrical connection, and commands the actuator to open one of the valve bodies, open the third valve body, close (or open) the second valve body, close the fourth valve body, compress the pump body to drive the gas in the pump body through the third valve body (or, and the second valve body), and then discharge it through the fluid inlet.

[0092] The main control board commands to stop compressing the pump body, keeps valve body two closed (or closed), and commands the electrical module to apply a weak electrical signal between the two electrical contacts corresponding to electrode one and electrode two. The fluid inlet is exposed to the environment, and the blood sample to be tested is brought into contact with the fluid inlet. The first countdown of the timer starts, and at the same time, the main control board commands to release the pump body to drive the blood sample into the microfluidic channel. During the process of the blood sample to be tested entering the microfluidic channel, part of the gas enters the pump body through valve body three, and the volume of the gas entering the pump body is proportional to the volume of the blood sample to be tested that enters.

[0093] When the first countdown of the timer ends, the blood sample stops entering, the main control board determines that the sample addition has been successful, and commands the actuator to close valve body one, open valve body two, close valve body three, open valve body four, and compress the pump body to drive the blood sample to be tested between valve body two and valve body three into the reaction area. In the process of the blood sample to be tested entering the reaction area, part of the 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 that enters.

[0094] The front end of the blood sample to be tested contacts the termination end of the reaction zone, and a conductive path is formed between the electrode sensing end 1 and the electrode sensing end 2 through the blood sample to be tested. The electrical signal can be detected through the electrode contact 1 and the electrode contact 2, indicating that the microfluidic sample has moved to the reaction zone. 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 a weak electrical signal between the two electrical contact pins corresponding to the electrode contact 1 and the electrode contact 2, waiting for the blood sample to be tested to complete the reaction with the preset reagent in the reaction zone and electron transfer to occur. The current representing the reaction result signal is generated by the electrode sensing end. The electrons are received at the electrode sensing end 1 and the electrode sensing end 2, and are transmitted to the detection device through the electrode contact 1, the electrode contact 2, the electrical contact pin 1, and the electrode contact pin 2. The main control board receives and detects the current generated by the electron transfer, processes and determines the concentration of β-hydroxybutyric acid in the blood sample and commands the display screen to display the detection result. At the same time, the actuator is commanded to restore the compression pump body to drive the waste liquid after the reaction to enter the liquid collecting area. When the waste liquid after the reaction enters the liquid collecting area, part of the gas is discharged through the fluid outlet. The liquid collecting area absorbs the waste liquid after the reaction. The used microfluidic sample detection device is taken out from the detection device and discarded.

[0095] Example 3

[0096] The present invention is implemented on a microfluidic sample detection system composed of the third microfluidic detection device and the microfluidic detection equipment.

[0097] The difference from other embodiments is that when the sample to be tested contacts the fluid inlet, on the one hand, it first flows toward the branch outlet, and on the other hand, as the pump body is released, branch one of the sample to be tested flows toward valve body two and valve body three, and then stops flowing when the pump body stops releasing; or, as the pump body is released, branch one of the sample to be tested first flows toward valve body two and valve body three, and then as the pump body is compressed, one of the branches of the sample to be tested continues to flow downstream, and then stops flowing when the pump body stops compressing, while branch two of the sample to be tested that first flows toward the branch outlet stops flowing when the front end of the sample to be tested approaches or reaches the branch outlet.

[0098] The difference from other embodiments is that the main control board commands the release of the pump body to drive the blood sample into the microfluidic channel and commands the electrical module to apply a weak electrical signal between the two electrical contact pins corresponding to the electrode three and the electrode four, and the sample to be tested is connected with the electrode sensing end three and the electrode sensing end four in the microfluidic channel branch, so that a loop is formed between the electrode three and the electrode four to determine that the sample addition is successful. In another embodiment, the interference compensation operation is performed by capturing the electrical signal between the electrode sensing end three and the electrode sensing end four.

[0099] Example 4

[0100] The present invention is implemented on a microfluidic sample detection system composed of the fourth microfluidic detection device and the microfluidic detection equipment.

[0101] The difference from other embodiments is that the pre-set reagent in liquid form is framed in the microfluidic channel within the electrode frame area, and when the microfluid sample flows into the electrode frame area, it reacts with the pre-set reagent in liquid form. Furthermore, the pre-set reagent in liquid form is framed in the microfluidic channel within the electrode frame area, and when the microfluid sample flows into the electrode frame area, it reacts with the pre-set reagent in liquid form, and a signal change is generated between the frame electrode and another electrode within the electrode frame area, and is transmitted to the microfluid detection instrument through the frame contact and the contact of the other electrode.

[0102] Example 5

[0103] The present invention is implemented on a microfluidic sample detection system composed of the microfluidic detection device No. 5 and the microfluidic detection equipment.

[0104] Red blood cells are used as the first target, and the reagents pre-installed in area one are magnetic particles combined with anti-red blood cell antibodies. β-hydroxybutyrate is used as the second target, and the reagents pre-installed in area two are preparations containing β-hydroxybutyrate oxidoreductase and an electron mediator. In this case, both area one and area two are reaction areas.

[0105] Turn on the power of the detection device, take the microfluidic detection device and put it into the load position of the microfluidic detection device, the electrical contact pins in the detection device are paired with electrode contact 1, electrode contact 2, and electrode contact 5, and the load position generates a loaded signal and transmits it to the main control board, indicating that the detection device and the detection device have completed the electrode electrical connection. In another embodiment, the load position does not generate a loaded signal, but transmits a signal that the electrodes are electrically connected to the main control board by manually clicking a button on the detection device.

[0106] The main control board receives the signal of the electrical connection of the electrodes, and commands the actuator to open valve body one, valve body three, valve body two, close valve body four, compress the pump body to drive the gas in the pump body through valve body three and valve body two, and then discharge it through the fluid inlet.

[0107] The main control board commands to stop compressing the pump body, and the fluid inlet is exposed to the environment. The blood sample to be tested is brought into contact with the fluid inlet, and the first countdown of the timer starts. At the same time, the main control board commands to release the pump body to drive the blood sample to be tested into the microfluidic channel. During the process of the blood sample to be tested entering the microfluidic channel, part of the gas enters the pump body through valve body two and valve body three. The volume of gas entering the pump body is proportional to the volume of the blood sample to be tested that enters. When the front end of the blood sample to be tested approaches valve body two, the main control board commands to close valve body two, but continues to keep valve body three open, and the actuator continues to release the pump body, and the blood sample to be tested continues to flow downstream of valve body two.

[0108] When the first countdown of the timer ends, the blood sample to be tested stops entering, the main control board determines that the sample addition has been successful, and commands the electrical module to apply a weak electrical signal to the two electrical contacts corresponding to electrode one and electrode two, and commands the actuator to close valve body one, keep valve body two closed, and continue to release the pump body to drive the blood sample to be tested between valve body one and valve body three to enter area one. However, when the blood sample does not fill area one and the front end of the blood sample approaches valve body three, the main control board commands the actuator to close valve body three, open valve body two, open valve body four, and compress the pump body to drive the blood sample to be tested between valve body two and valve body three to continue to flow to fill area one.

[0109] The front end of the blood sample to be tested contacts the termination end of area 1, that is, contacts the electrode sensing end 2. A conductive path is formed between the electrode sensing end 1 and the electrode sensing end 2 through the blood sample to be tested. The electrical signal can be detected through the electrode contact 1 and the electrode contact 2. 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 a weak electrical signal between the two electrical contact pins corresponding to the electrode contact 1 and the electrode contact 2. The second countdown of the timer starts, waiting for the blood sample to be tested to react with the magnetic particles in area 1 and the anti-red blood cell antibodies. The red blood cells in the blood sample to be tested combine with the antibodies to form a magnetic particle-anti-red blood cell antibody-red blood cell complex. When the second countdown of the timer ends, the main control board commands the magnetic shielding body in the magnetic module to move and leave the space between the magnet and the microfluidic channel, so that area one is exposed to the magnetic field generated by the magnet. The red blood cells bound to the antibody are adsorbed on the wall of the microfluidic channel in area one due to the effect of the magnetic field. The main control board commands the actuator to continue compressing the pump body to drive the plasma without red blood cells to flow from area one into area 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 five. During the process, part of the gas is discharged through the fluid outlet, and the volume of the discharged gas is equivalent to the volume of the plasma sample without red blood cells entering area two.

[0110] The front end of the plasma contacts the second termination end of the area, and a conductive path is formed between the electrode sensing end 2 and the electrode sensing end 5 through the blood sample to be tested. The electrical signal can be detected through the electrode contact 2 and the electrode contact 5. 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 a weak electrical signal between the two electrical contact pins corresponding to the electrode contact 2 and the electrode contact 5, waiting for the plasma to react with the preparation containing β-hydroxybutyrate oxidoreductase and the electron mediator to complete and electron transfer to occur. The current is transmitted to the detection device through the electrode sensing end 2, the electrode sensing end 5, the electrode contact 2, the electrode contact 5, the electrical contact pin 2, and the electrode contact pin 5. The main control board receives the detected current, processes and determines the β-hydroxybutyric acid concentration in the plasma and commands the display screen to display the detection result. At the same time, the actuator is commanded to resume compressing the pump body to drive the waste liquid after the reaction into the liquid collection area. When the waste liquid after the reaction enters the liquid collection area, part of the gas is discharged through the fluid outlet. The liquid collection area absorbs the waste liquid after the reaction, and the used microfluidic sample detection device is taken out from the detection device and discarded.

[0111] The above test was repeated 10 times with blood samples with β-hydroxybutyric acid content of 1mmol / L, 2mmol / L, 4mmol / L, 6mmol / L, and 7mmol / L, and the test results were obtained. A biochemical standard instrument was used as a control, and the same blood samples were tested 10 times on the existing brand A traditional electrochemical blood glucose detection products with excellent performance and brand B traditional electrochemical blood glucose detection products with average performance, as shown in Table 1. As can be seen from Table 1, the standard deviation and coefficient of variation are significantly reduced compared with both the traditional brand B products and the traditional brand A products, indicating that the test result data of the present invention has a lower degree of discreteness, and the results of measuring the β-hydroxybutyric acid content in the blood samples have less repeatability. The data in Table 1 were studied for accuracy and plotted, as shown in Table 1. Fig. 9 , Fig.10 and Fig.11 As shown, R 2 Significantly greater than the B traditional brand product with average performance, and greater than the R traditional brand product with excellent performance 2 This indicates that the results of the present invention for measuring the content of β-hydroxybutyric acid in blood samples are closer to the true value, and the accuracy is even higher than that of existing products with excellent performance.

[0112] Table 1 Comparative experimental results of Example 6

[0113]

[0114] Example 6

[0115] The present invention is implemented on a microfluidic sample detection system composed of the microfluidic detection device six and the microfluidic detection equipment.

[0116] C-reactive protein is used as the target. The reagent pre-installed in area 1 is C-reactive protein antibody combined with magnetic particles, that is, C-reactive protein antibody labeled with magnetic particles. The reagent pre-installed in area 2 is C-reactive protein antibody labeled with fluorescent particles. There is no pre-installed reagent in area 3. In this case, area 1 and area 2 are reaction areas, and area 3 is the detection area.

[0117] Turn on the power of the detection device, take the microfluidic detection device and put it into the load position of the microfluidic detection device, the electrical contact pins in the detection device are paired with electrode contact one, electrode contact two, electrode contact five, and electrode contact six, and the load position generates a signal of already loaded and transmits it to the main control board, indicating that the detection device and the detection device have completed the electrical connection of the electrodes. In another embodiment, the load position does not generate a signal of already loaded, but transmits a signal of already electrically connected electrodes to the main control board by manually clicking a button on the detection device. The main control board receives the signal of electrically connected electrodes, and commands the actuator to open one of the valve bodies, open the third valve body, close the second valve body, close the fourth valve body, compress the pump body to drive the gas in the pump body through the third valve body, and then discharge it through the fluid inlet.

[0118] The main control board commands to stop compressing the pump body, open the second valve body, expose the fluid inlet to the environment, and bring the blood sample to be tested into contact with the fluid inlet. The first countdown of the timer starts, and at the same time, the main control board commands to release the pump body to drive the blood sample to be tested into the microfluidic channel. The blood sample to be tested enters the microfluidic channel. When the front end of the blood sample to be tested approaches the second valve body, the main control board commands to close the second valve body, but continues to keep the third valve body open, and the actuator continues to release the pump body, and the blood sample to be tested continues to flow downstream of the second valve body.

[0119] After the first countdown of the timer ends, the main control board determines that the sample addition has been successful, and instructs the electrical module to apply a weak electrical signal to the two electrical contacts corresponding to electrode one and electrode two, and instructs the actuator to close valve body one and continue to release the pump body to drive the blood sample to be tested between valve body one and valve body three to enter area one. However, when the blood sample does not fill area one and the front end of the blood sample approaches valve body three, the main control board instructs the actuator to close valve body three, open valve body two, open valve body four, and compress the pump body to drive the blood sample to be tested between valve body two and valve body three to continue to flow to fill area one.

[0120] The front end of the blood sample to be tested contacts the termination end of area one, and a conductive path is formed between the electrode sensing end one and the electrode sensing end two through the blood sample to be tested. The electrical signal can be detected through the electrode contact one and the electrode contact two. The main control board commands the actuator to stop compressing the pump body, and at the same time commands the electrical module to stop applying a weak electrical signal between the two electrical contact pins corresponding to the electrode contact one and the electrode contact two, and between the two electrical contact pins corresponding to the electrode contact two and the electrode contact six. The second countdown of the timer starts, waiting for the blood sample to be tested to react with the magnetic particles combined with the C-reactive protein antibody in area one, and the C-reactive protein in the blood sample to be tested combines with the antibody to form a magnetic particle C-reactive protein antibody-C-reactive protein complex. After 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 to flow from area one into area two and contact the terminal end of area two, that is, a conductive path is formed between the electrode sensing end two and the electrode sensing end six through the blood sample containing the aforementioned complex, and the electrical signal can be detected through the electrode contact two and the 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 a weak electrical signal between the two electrical contact pins corresponding to the electrode contact two and the electrode contact six. The third countdown of the timer begins, waiting for the blood sample containing the aforementioned complex to combine with the C-reactive protein antibody labeled with fluorescent particles to form a magnetic particle C-reactive protein antibody-C-reactive protein-fluorescent particle labeled C-reactive protein antibody complex, that is, a double antibody sandwich complex. When the third countdown of the timer ends, the main control board commands the actuator to resume compressing the pump body to drive the micro blood sample containing the aforementioned complex to flow from area two into area three and contact the terminal end of area three, and commands the electrical module to apply a weak electrical signal between the two electrical contact pins corresponding to electrode contact six and electrode contact five. After that, a conductive path is formed between electrode sensing end two and electrode sensing end six through the blood sample containing the aforementioned complex, and the electrical signal can be detected through electrode contact six and electrode contact five. 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 contact pins corresponding to electrode contact six and electrode contact five. The main control board commands the magnetic shield in the magnetic module to move so as to leave the magnet and The microfluidic channels are connected so that area 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 on the microfluidic channel wall of area three due to the effect of the magnetic field. The main control board commands the light detection unit in the optical detection module to perform fluorescent signal detection on the complex in area three. The main control board receives the detected fluorescent signal intensity, and the photoelectric conversion unit converts the fluorescent signal intensity into an electrical signal value. The main control board receives the detected electrical signal value, converts the C-reactive protein content in the blood sample and commands the display screen to display the result. At the same time, the main control board commands the actuator to resume the compression pump body to drive the waste liquid that has lost the aforementioned complex from area three into the liquid collection area.

[0121] The liquid collecting area absorbs the waste liquid, takes out the used microfluidic sample detection device from the detection device and discards it. In this embodiment, the reagents preset in area 1 and area 2 can be exchanged with each other.

[0122] In another embodiment, determining the travel area of ​​the microfluid sample by the pressure value applied by the actuator on the pump body can replace the role of applying a weak electrical signal between the two electrode sensing ends to determine whether the microfluid sample reaches the area corresponding to the two electrode sensing ends.

[0123] Example 7

[0124] 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.

[0125] In this case, the electrode sensing end is a reference Figure 1-2 As shown in (a), in the middle of valve body two and valve body three, area one is the pretreatment area, area two and area three are the reaction areas, and area four is the detection area.

[0126] Total vitamin D (total 25-hydroxyvitamin D) is the target, the reagent preset in area 1 is VD dissociation solution, the reagent preset in area 2 is the antibody of magnetic particle binding first epitope, that is, the first epitope antibody labeled by magnetic particle, the reagent preset in area 3 is the second epitope antibody that can be labeled by fluorescein or fluorescent particle, and there is no preset reagent in area 4. Electrode sensing end 5 is located at the starting end of area 4, and electrode sensing end 7 is located at the ending end of area 4.

[0127] Turn on the power of the detection device, take the microfluidic detection device and put it into the load position of the microfluidic detection device, the electrical contact pins in the detection device are paired with electrode contact 1, electrode contact 2, electrode contact 5, electrode contact 6, and electrode contact 7, and the load position generates a loaded signal and transmits it to the main control board, indicating that the detection device and the detection device have completed the electrode electrical connection. In another embodiment, the load position does not generate a loaded signal, but transmits a signal that the electrodes are electrically connected to the main control board by manually clicking a button on the detection device.

[0128] The main control board receives the signal of the electrical connection of the electrodes, and instructs the actuator to open valve body one, valve body three, valve body two, close valve body four, compress the pump body to drive the gas in the pump body through valve body three, and then discharge it through the fluid inlet.

[0129] The main control board commands to stop compressing the pump body, and the fluid inlet is exposed to the environment. The blood sample to be tested is brought into contact with the fluid inlet, and the first countdown of the timer starts. At the same time, the main control board commands to release the pump body to drive the blood sample to be tested into the microfluidic channel. The blood sample to be tested enters the microfluidic channel. When the front end of the blood sample to be tested approaches the second valve body, the main control board commands to close the second valve body, but continues to keep the third valve body open, and the actuator continues to release the pump body, and the blood sample to be tested continues to flow downstream of the second valve body.

[0130] After the first countdown of the timer ends, the main control board determines that the sample addition has been successful, and instructs the electrical module to apply a weak electrical signal to the two electrical contacts corresponding to electrode one and electrode two, instructing the actuator to close one of the valve bodies and continue to release the pump body to drive the blood sample into area one.

[0131] The front end of the blood sample to be tested contacts the termination end of area 1, and a conductive path is formed between the electrode sensing end 1 and the electrode sensing end 2 through the blood sample to be tested. The electrical signal can be detected through the electrode contact 1 and the electrode contact 2. The main control board commands the actuator to stop releasing the pump body, and at the same time commands the electrical module to stop applying a weak electrical signal between the two electrical contact pins corresponding to the electrode contact 1 and the electrode contact 2. The second countdown of the timer starts, waiting for the blood sample to be tested to be fully incubated and dissociated by the VD dissociation solution in area 1. In order to fully release the bound VD in the blood sample to be tested, the main control board can also command the temperature module to heat area 1 before the second countdown of the timer starts. Dissociating VD from the blood sample is a specific treatment of the bound VD. Only by specifically dissociating it from the blood can the subsequent binding reaction be realized to determine the total VD content in the blood. After the second countdown of the timer ends, the main control board commands the electrical module to start applying a weak electrical signal between the two electrical contact pins corresponding to electrode contact 2 and electrode contact 6, and commands the actuator to close valve body 3, open valve body 1, and compress the pump body to drive the blood sample to be tested containing the dissociated state VD to flow from area 1 to area 2 and contact the terminal end of area 2, that is, a conductive path is formed between electrode sensing end 2 and electrode sensing end 6 through the blood sample to be tested containing the dissociated state VD, and the electrical signal can be detected through electrode contact 2 and electrode contact 6. 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 a weak electrical signal between the two electrical contact pins corresponding to electrode contact 2 and electrode contact 6. The third countdown of the timer starts, waiting for the blood sample to be tested containing the dissociated state VD to react with the first epitope antibody labeled with magnetic particles, and the dissociated state VD in the blood sample to be tested combines with the antibody to form a magnetic particle VD antibody-VD antigen complex. After the third 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 to flow from area two into area three and contact the terminal end of area three, and commands the electrical module to apply a weak electrical signal between the two electrical contact pins corresponding to electrode contact six and electrode contact five. A conductive path is formed between electrode sensing end six and electrode sensing end five through the blood sample containing the aforementioned complex, and the electrical signal can be detected through electrode contact six and electrode contact five. 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 a weak electrical signal between the two electrical contact pins corresponding to electrode contact six and electrode contact five. The fourth countdown of the timer begins, waiting for the blood sample containing the aforementioned complex to bind to the second epitope antibody labeled with the fluorescent particles to form a magnetic particle VD antibody-VD antigen-fluorescent particle VD antibody complex, that is, a double antibody sandwich complex.After the fourth countdown of the timer ends, the main control board commands the actuator to resume compressing the pump body to drive the micro blood sample containing the aforementioned complex to flow from area three into area four and contact the terminal end of area four, and commands the electrical module to apply a weak electrical signal between the two electrical contact pins corresponding to electrode contact five and electrode contact seven. A conductive path is formed between electrode sensing end five and electrode sensing end seven through the blood sample containing the aforementioned complex, and the electrical signal can be detected through electrode contact five and electrode contact seven. 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 contact pins corresponding to electrode contact five and electrode contact seven. The main control board commands the magnetic shield in the magnetic module to move so that Leave the area between the magnet and the microfluidic channel so that area 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 on the microfluidic channel wall of area four due to the effect of the magnetic field. The main control board commands the light detection unit in the optical detection module to detect the fluorescent signal of the complex in area four. The main control board receives the detected fluorescent signal intensity, and the photoelectric conversion unit converts the fluorescent signal intensity into an electrical signal value. The main control board receives the detected electrical signal value, converts 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 resume the compression pump body to drive the waste liquid that has lost the aforementioned complex from area three into the liquid collection area.

[0132] The liquid collecting area absorbs the waste liquid, takes out the used microfluidic sample detection device from the detection device and discards it. In this embodiment, the reagents preset in area 2 and area 3 can be exchanged with each other.

[0133] In another embodiment, determining the travel area of ​​the microfluid sample by the pressure value applied by the actuator on the pump body can replace the role of applying a weak electrical signal between the two electrode sensing ends to determine whether the microfluid sample reaches the area corresponding to the two electrode sensing ends.

[0134] Example 8

[0135] The present invention is implemented in a microfluidic sample detection system composed of the microfluidic detection device No. 7 and the microfluidic detection equipment.

[0136] The difference from other embodiments is that the pre-set reagent in liquid form is framed in the microfluidic channel within the electrode frame area, and when the microfluid sample flows into the electrode frame area, it reacts with the pre-set reagent in liquid form. Furthermore, the pre-set reagent in liquid form is framed in the microfluidic channel within the electrode frame area, and when the microfluid sample flows into the electrode frame area, it reacts with the pre-set reagent in liquid form, and a signal change is generated between the frame electrode and the second electrode, and is transmitted to the microfluid detection instrument through the frame contact and the second electrode contact.

[0137] Example 9

[0138] The present invention is implemented on a microfluidic sample detection system composed of the microfluidic detection device No. 8 and the microfluidic detection equipment.

[0139] The difference from other embodiments is that the detection device includes an electrical contact pin paired with electrode contact one and electrode contact eight, and the main control board commands the release of the pump body to drive the microfluid sample to be tested into the microfluid channel, and at the same time commands the electrical module to apply a weak electrical signal to electrode one and electrode eight through the contact pin. When the front end of the microfluid sample contacts the electrode sensing end one, a conductive path is formed between the electrode sensing end one and the electrode sensing end eight through the microfluidic sample, and the electrical signal can be detected through electrode contact one and electrode contact eight. The main control board determines that the microfluidic sample is successfully added, and the microfluidic sample in the microfluidic channel from valve body two to electrode sensing end one is a determined microfluidic sample volume.

[0140] Example 10

[0141] The present invention is implemented on a microfluid sample detection system composed of a combined microfluid detection device and a microfluid detection equipment.

[0142] A device for simultaneously detecting glucose, β-hydroxybutyrate and hemoglobin in blood, wherein device unit one 101 is used for detecting glucose, device unit two 102 is used for detecting β-hydroxybutyrate, device unit three 104 and device unit four are used for detecting hemoglobin 105.

[0143] In order to expand the signal response of β-hydroxybutyric acid, the second electrode sensing end 1021 of unit 2 is thickened. In order to make the area where the sample to be tested stays and the area that reacts with the preset reagent overlap, the horizontal edge of the electrode frame area 1 1032 in the device unit 3 overlaps with the electrode sensing end 1031 of unit 3, and the horizontal edge of the electrode frame area 2 1042 in the device unit 4 overlaps with the electrode sensing end 1041 of unit 4.

[0144] Each of the device units 1 to 4 is composed of two regions of electrode sensing ends of three electrodes. The reagents pre-installed in the two regions of the device unit 1 are, from bottom to top, one, magnetic particles combined with anti-erythrocyte antibodies, and the other, a preparation containing glucose oxidoreductase and an electron mediator; the reagents pre-installed in the two regions of the device unit 2 are, from bottom to top, one, magnetic particles combined with anti-erythrocyte antibodies, and the other, a preparation containing β-hydroxybutyrate oxidoreductase and an electron mediator; the reagents pre-installed in the two regions of the device unit 3 and the device unit 4 are, from bottom to top, one, hemoglobin antibodies combined with magnetic particles, and the other, hemoglobin antibodies labeled with fluorescent particles; the device unit 4 and the device unit 3 are a control channel and a detection channel, so that errors can be eliminated through the control channel while detecting the hemoglobin content.

[0145] The detection process of device unit 1 and device unit 2 is the same as that of Example 6. The detection process of device unit 3 and device unit 4 is similar to that of Example 8, except that after the formation of the double antibody sandwich complex and the end of the third countdown of the timer, the main control board commands the magnetic shielding body in the magnetic module to move so as to leave between the magnet and the microfluidic channel, so that area 2 is exposed to the magnetic field generated by the magnet, and the aforementioned double antibody sandwich complex is adsorbed on the microfluidic channel wall of area 2 due to the action of the magnetic field, and the main control board commands the light detection unit in the optical detection module to perform fluorescence signal detection on the complex in area 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, converts the hemoglobin content in the blood sample and commands the display screen to display the result, and 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 area 2 into the liquid collection area.

[0146] Embodiment 11

[0147] The present invention is implemented on a system composed of one of the microfluidic detection devices and a microfluidic detection equipment, and the microfluidic sample detection method of the present invention is used to separate red blood cells and plasma in a blood sample and extract plasma. Red blood cells are used as targets, and the reagents pre-set in the reaction area are magnetic particles combined with anti-red blood cell antibodies.

[0148] Step 1: Turn on the power of the detection device, take the microfluidic detection device and put it on the load position of the microfluidic detection device, and the fluid inlet is exposed to the environment. The load position generates a loaded signal and transmits it to the main control board. In another embodiment, the load position does not generate a loaded signal, but transmits the loaded signal to the main control board by manually clicking the button on the detection device. The microfluidic detection device is connected to the microfluidic detection device signal, and the main control board commands the actuator to open one of the valve bodies, open the third valve body, close (or open) the second valve body, close the fourth valve body, compress the pump body to drive the gas in the pump body through the third valve body (or, and the second valve body), and then discharge it through the fluid inlet. The main control board commands to stop compressing the pump body, keep the second valve body closed (or closed), contact the blood sample with the fluid inlet, and the first countdown of the timer begins.

[0149] Step 2: The main control board commands the release of the pump body to drive the blood sample into the microfluidic channel. During the process of the blood sample entering the microfluidic channel, part of the gas enters the pump body through the third valve body. The volume of gas entering the pump body is proportional to the volume of the blood sample entering.

[0150] Step 3: At the same time, the main control board commands to stop releasing the pump body, and the blood sample stops entering. The main control board determines that the sample addition has been successful, and then commands the actuator to close valve body one, open (or keep open) valve body two, close valve body three, open valve body four, compress the pump body to drive the blood sample between valve body two and valve body three into the reaction area, and the second countdown of the timer begins. In the process of the blood sample entering the reaction area, part of the gas is discharged through the fluid outlet, and the volume of the discharged gas is proportional to the volume of the blood sample traveling downstream.

[0151] Step 4: 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 compressing the pump body, and the third countdown of the timer begins.

[0152] Step 5: The blood sample reacts with the anti-erythrocyte antibody combined with the magnetic particles in the reaction area. The red blood cells in the blood sample combine with the antibody to form a magnetic particle-anti-erythrocyte antibody-erythrocyte complex.

[0153] Step 6: After the third countdown of the timer ends, the main control board commands the magnetic shield in the magnetic module to move away from between the magnet and the microfluidic channel, so that the reaction area is exposed to the magnetic field generated by the magnet.

[0154] Step 7: The red blood cells bound to the antibodies are adsorbed on the wall of the microfluidic channel in the reaction zone due to the effect of the magnetic field. The main control board commands the actuator to continue compressing the pump body to drive the plasma that has lost the red blood cells to leave the reaction zone and flow into the liquid collection area and out of the fluid outlet for subsequent use.

[0155] The operation sequence between the above steps does not have to be completely followed. For example, the operation at the end of the first step and the operation at the beginning of the second step are performed at the same time.

[0156] In another embodiment, in the first step, after the gas in the pump body is discharged, the main control board commands to stop compressing the pump body, open (or keep open) the second valve body, and contact the blood sample with the fluid inlet, and the first countdown of the timer begins. In the second step, the main control board commands to release the pump body to drive the blood sample into the microfluidic channel. During this process, part of the gas enters the pump body through the second valve body and the third valve body. The volume of the gas entering the pump body is proportional to the volume of the blood sample entering. When the front end of the blood sample approaches the second valve body, the main control board commands to close the second valve body, but continues to keep the third valve body open.

[0157] In another embodiment, there is no first countdown, and the optical module determines that the blood sample is successfully added; in another embodiment, there is no second countdown, and the pressure value applied by the actuator on the pump body determines that the blood sample has filled the reaction area.

[0158] In a further embodiment, one of the above microfluidic detection devices is replaced by a second microfluidic detection device, the difference being that:

[0159] Detection device such as Figure 1-2 In the case shown in (b), in the third step, after the sample is added successfully, the main control board commands the electrical module to apply a weak electrical signal to the two electrical contacts corresponding to electrode one and electrode two. In the fourth step, the front end of the blood sample contacts electrode sensing end two, and a conductive path is formed between electrode sensing end one and electrode sensing end two through the blood sample to be tested. The electrical signal can be detected by electrode contact one and electrode contact two. The main control board commands the actuator to stop compressing the pump body, and at the same time commands the electrical module to stop applying a weak signal between the two electrical contacts corresponding to electrode contact one and electrode contact two. There is no need for a second countdown function in the timer. If the detection device is as follows Figure 1-2 (c) and the electrode sensing end 1 is closer to the valve body 3, the implementation is the same.

[0160] Detection device such as Figure 1-2In the case shown in (a), in the third step, after the sample is added successfully, the main control board commands the electrical module to apply a weak electrical signal to the two electrical contacts corresponding to electrode one and electrode two, and commands the actuator to close one of the valve bodies, keep valve two closed (or close), and continue to release the pump body to drive the blood sample into the reaction area. In the fourth step, the front end of the blood sample contacts electrode sensing end two, and a conductive path is formed between electrode sensing end one and electrode sensing end two through the blood sample to be tested. The electrical signal can be detected through electrode contact one and electrode contact two. The main control board commands the actuator to stop releasing the pump body, and at the same time commands the electrical module to stop applying a weak signal between the two electrical contacts corresponding to electrode contact one and electrode contact two. Similarly, there is no need for a second countdown function in the timer. If the detection device is as follows: Figure 1-2 (c) and the electrode sensing end 1 is closer to the valve body 2, the implementation is the same.

[0161] When the detection device is Figure 1-2 (c) and when the electrode sensing end 1 is between valve body 2 and valve body 3, in the third step, after the sample is added successfully, the main control board commands the electrical module to apply a weak electrical signal to the two electrical contacts corresponding to electrode 1 and electrode 2, and commands the actuator to close valve body 1, keep valve body 2 closed (or closed), and continue to release the pump body to drive the blood sample into the reaction area. However, when the blood sample does not fill the reaction area and the front end of the blood sample is close to valve body 3, the main control board commands the actuator to close valve body 3, open valve body 2, open valve body 4, and compress the pump body to drive the blood sample between valve body 2 and valve body 3 to continue to flow to fill the reaction area. In the fourth step, the front end of the blood sample contacts the electrode sensing terminal 2, and the blood sample fills the reaction area. A conductive path is formed between the electrode sensing terminal 1 and the electrode sensing terminal 2 through the blood sample to be tested. The electrical signal can be detected through the electrode contact 1 and the electrode contact 2. 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 a weak signal between the two electrical contact pins corresponding to the electrode contact 1 and the electrode contact 2. Similarly, there is no need for a second countdown function in the timer.

[0162] The above embodiments are only used to more clearly illustrate the technical solutions of the present invention, and are therefore only examples, and cannot be used to limit the protection scope of the present invention.

[0163] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by technicians in the technical field to which the present invention belongs; the terms used herein are only for the purpose of describing specific embodiments and are not intended to limit the present invention; the terms "including" and "having" in the description and claims of the present invention and any variations thereof are intended to cover non-exclusive inclusions.

[0164] In the description of the present invention, the technical terms "first", "second", "third", "fourth", etc. are only used to distinguish different objects, and cannot be understood as indicating or implying relative importance or implicitly indicating the number, specific order or primary and secondary relationship of the indicated technical features. In the description of the embodiments of the present invention, the meaning of "multiple" is more than two, unless otherwise clearly and specifically defined.

[0165] References to "embodiments" and "implementations" herein mean that a particular feature, structure, or characteristic described in conjunction with the embodiments may be included in at least one embodiment of the present invention. The appearance of the phrase in various locations in the specification does not necessarily refer to the same embodiment, nor is it an independent or alternative embodiment that is mutually exclusive with other embodiments.

[0166] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit them; although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein by equivalents; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention, and they should all be included in 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 various embodiments can be combined in any way. The present invention is not limited to the specific embodiments disclosed herein, but includes all technical solutions that fall within the scope of the claims.

Claims

1. A microfluid sample detection method, characterized in that: include: Open the microfluidic detection device and place the microfluidic detection device in the load position, wherein the microfluidic detection device is signal-connected to the microfluidic detection device; The actuator opens the first valve body and the third valve body, closes the second valve body and the fourth valve body, and compresses the first pump body to drive at least part of the gas in the first pump body to be discharged from the fluid inlet through the third valve body, or opens the first valve body, the second valve body and the third valve body, closes the fourth valve body, compresses the first pump body to drive at least part of the gas in the first pump body to be discharged from the fluid inlet through the second valve body and the third valve body, and then closes the second valve body; In response to a signal that the microfluid sample contacts the fluid inlet, the main control board commands the release of the first pump body to drive the microfluid sample into the microfluid channel. After determining that the sample addition is successful, the main control board commands to stop releasing the first pump body and close the first valve body. The actuator executes the action of the first pump body to drive the microfluid sample into the reaction area, and closes the third valve body, opens the second valve body, and opens the fourth valve body before or after executing the action. The actuator stops the action of the first pump body, waits for the countdown to end, and compresses the first pump body to drive the microfluid sample after the reaction to continue to flow downstream into the liquid collection area, and detects the reaction result signal of the reaction area.

2. A microfluidic sample detection method according to claim 1, characterized in that: After the main control board determines that the sample loading is successful, it commands to stop releasing the first pump body and close the first valve body. Then, the actuator releases the first pump body to drive the microfluid sample into the reaction area. Then, the actuator closes the third valve body and opens the second valve body and the fourth valve body.

3. A microfluidic sample detection method according to claim 2, characterized in that: After the actuator releases the first pump body, it opens the second valve body before the microfluid sample fills the reaction area, and compresses the first pump body to drive the microfluid sample to fill the reaction area.

4. A microfluidic sample detection method according to claim 1, characterized in that: After the main control board determines that the sample loading is successful, it commands to stop releasing the first pump body, close the first valve body and the third valve body, and open the second valve body and the fourth valve body. Then, the actuator compresses the first pump body to drive the microfluid sample into the reaction area.

5. A microfluidic sample detection method according to claim 1, characterized in that: After the microfluidic detection device is connected to the microfluidic detection equipment by signal, the actuator opens the first valve body, the second valve body and the third valve body, closes the fourth valve body, closes the second valve body before driving the microfluidic sample into the reaction area, and the actuator releases the first pump body to drive the microfluidic sample into the reaction area.

6. A microfluidic sample detection method according to claim 5, characterized in that: After the actuator releases the first pump body, it opens the second valve body before the microfluid sample fills the reaction area, and compresses the first pump body to drive the microfluid sample to fill the reaction area.

7. A microfluidic sample detection method according to claims 1-6, characterized in that: The actuator performs an action of compressing the first pump body to drive the microfluid sample from the first area into the second area, and the countdown occurs before the microfluid sample enters the second area.

8. A microfluidic sample detection method according to any one of claims 1 to 6, characterized in that: The light detection unit determines the travel area of ​​the microfluid sample by continuously detecting the light reflection state.

9. A microfluidic sample detection method according to any one of claims 1 to 6, characterized in that: The travel area of ​​the microfluid sample is determined by the pressure value applied by the actuator to the first pump body.

10. A microfluidic sample detection method according to any one of claims 1 to 6, characterized in that: The microfluidic detection device is opened and the microfluidic detection device is placed in the load position, the first electrode contact and the second electrode contact are matched with the electrical contact pins respectively, and the microfluidic detection device and the microfluidic detection device electrodes are electrically connected.

11. A microfluid sample detection method according to claim 10, characterized in that: The electrical module determines the travel area of ​​the microfluid sample through the first electrode sensing end and the second electrode sensing end.

12. A microfluid sample detection method according to claim 10, 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 the reaction result signal to the microfluidic detection device through the first electrode contact, the second electrode contact, and the electrical contact pin.

13. A microfluidic sample detection method according to any one of claims 1 to 6, characterized in that: The actuator performs an action of compressing the first pump body to drive the microfluid sample from the first area into the second area, and the countdown occurs before the microfluid sample enters the second area.

14. A microfluidic sample detection method according to claim 13, characterized in that: The microfluidic detection device is opened and placed in the load position, the first electrode contact, the second electrode contact and the third electrode contact are matched with the electrical contact pins respectively, and the microfluidic detection device and the microfluidic detection device electrodes are electrically connected.

15. A microfluidic sample detection method according to claim 14, characterized in that: The electrical module determines a travel area of ​​the microfluid sample through at least two of the first electrode sensing end, the second electrode sensing end, and the third electrode sensing end.

16. A microfluidic sample detection method according to claim 14, characterized in that: The countdown also occurs after the microfluidic sample enters the second area. 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 and the third electrode contact, as well as the electrical contact pin.

17. A microfluidic sample detection method according to any one of claims 1 to 6, characterized in that: The microfluid sample enters the first area, the second area and the third area in sequence, and the countdown occurs before the microfluid sample enters the third area.

18. A microfluidic sample detection method according to claim 17, characterized in that The first electrode contact, the second electrode contact, the third electrode contact and the fourth electrode contact are matched with the electrical contact pins respectively to realize the electrical connection between the microfluidic detection device and the electrodes of the microfluidic detection equipment.

19. A microfluidic sample detection method according to claim 18, characterized in that: The electrical module determines a travel area of ​​the microfluidic sample through at least two of the first electrode sensing end, the second electrode sensing end, the third electrode sensing end, and the fourth electrode sensing end.

20. The microfluid sample detection method according to claim 18, characterized in that: The countdown also occurs after the microfluidic sample enters the third area. 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 and the fourth electrode contact, as well as the electrical contact pin.

21. A microfluidic sample detection method according to any one of claims 1 to 6, characterized in that: The microfluid sample enters the first area, the second area, the third area and the fourth area in sequence, and the countdown occurs before the microfluid sample enters the fourth area.

22. A microfluidic sample detection method according to claim 21, characterized in that: 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 contact pins to achieve electrical connection between the microfluidic detection device and the electrodes of the microfluidic detection equipment.

23. A microfluid sample detection method according to claim 22, characterized in that: The electrical module determines a travel area of ​​the microfluidic sample through at least two of the first electrode sensing end, the second electrode sensing end, the third electrode sensing end, the fourth electrode sensing end, and the fifth electrode sensing end.

24. A microfluidic sample detection method according to claim 22, characterized in that: The countdown also occurs after the microfluidic sample enters the fourth area. 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 and the fifth electrode contact, as well as the electrical contact pin.

25. A microfluidic sample detection method according to any one of claims 1-6 or 11-12 or 14-16 or 18-20 or 22-24, characterized in that: The optical module obtains a reaction result signal by detecting the most downstream area.

26. A microfluid sample detection method according to any one of claims 1-6 or 11-12 or 14-16 or 18-20 or 22-24, characterized in that: The reaction area includes a target antibody labeled with magnetic particles, and the antibody can specifically bind to the target in the microfluidic sample.

27. A microfluidic sample detection method according to any one of claims 1-6 or 11-12 or 14-16 or 18-20 or 22-24, characterized in that: The reaction area includes antibodies labeled with fluorescent particles, and the antibodies specifically bind to targets in the microfluidic sample.

28. A microfluidic sample detection method according to any one of claims 1-6 or 11-12 or 14-16 or 18-20 or 22-24, characterized in that: The reaction area includes a first reaction area and a second reaction area, and a first target antibody labeled with magnetic particles and a second target antibody labeled with fluorescent particles are pre-set in the first reaction area and the second reaction area respectively; The actuator performs a first pump action to drive the microfluid sample to first enter the first reaction zone and then enter the second reaction zone, and the target object, the first antibody and the second antibody together form a double antibody sandwich complex; Before the actuator compresses the first pump body and drives the reacted microfluidic sample to continue to flow, the magnetic module generates a magnetic field to adsorb the double antibody sandwich complex on the wall of the microfluidic channel.

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