Microfluid sample detection device and microfluid sample detection system

Abstract

The invention provides a microfluid sample detection device and a microfluid sample detection system.The microfluid sample detection device comprises a fluid inlet, a microfluid channel, a fluid outlet, a reaction area arranged between the fluid inlet and the fluid outlet and partially coinciding with the microfluid channel, and a liquid collecting area arranged on the downstream of the reaction area, the first pump body is arranged at the upstream of the reaction area; the first valve body is arranged at the downstream of the fluid inlet and is used for controlling the connection and disconnection between the microfluid channel and the fluid inlet; the second valve body is arranged at the downstream of the first valve body and is used for controlling the connection and disconnection between the first pump body and the microfluid channel; a first lock body is disposed downstream of the second valve body and together with the second valve body defines a volume of the microfluidic sample. The pump body provides driving force for flowing of the microfluid sample when being communicated with the microfluid channel, and the working states of the first valve body and the second valve body are opposite. The microfluid sample detection device further comprises a conductive system composed of electrodes, and the conductive system at least comprises two electrodes.

Description

Technical Field

[0001] The present invention relates to the field of medical devices, and in particular to a microfluid sample detection device and a microfluid sample detection system. 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 microfluidic sample detection device, comprising a fluid inlet, a microfluidic channel and a fluid outlet, and:

[0006] a reaction zone, disposed between the fluid inlet and the fluid outlet and partially overlapping the microfluidic channel;

[0007] A liquid receiving zone is arranged downstream of the reaction zone, and the fluid outlet is the outlet of the liquid receiving zone;

[0008] A first pump body is disposed upstream of the reaction zone and provides a driving force for the flow of the microfluidic sample when communicating with the microfluidic channel;

[0009] A first valve body, disposed downstream of the fluid inlet and controlling the connection between the microfluidic channel and the fluid inlet;

[0010] A second valve body is disposed downstream of the first valve body and controls the on-off between the first pump body and the microfluidic channel. The working states of the first valve body and the second valve body are opposite.

[0011] The first lock body is disposed downstream of the second valve body and defines the volume of the microfluid sample together with the second valve body.

[0012] Furthermore, the microfluidic channel includes a first area and a second area, at least the first area or the second area is a reaction area, and the first pump body can drive the microfluidic sample to enter the first area and the second area in sequence.

[0013] Furthermore, the microfluidic channel includes a first area, a second area and a third area, at least the first area or the second area or the third area is a reaction area, and the first pump body can drive the microfluidic sample to enter the first area, the second area and the third area in sequence.

[0014] In another case, the microfluidic sample detection device also includes a conductive system composed of electrodes, the conductive system includes at least two electrodes, and the electrode sensing ends are respectively located at the two ends of the reaction zone. Preferably, the conductive system includes at least three electrodes, and the electrode sensing ends are distributed at the two ends of the first area and the second area, at least the first area or the second area is the reaction area, and the first pump body can drive the microfluidic sample to enter the first area and the second area in sequence. Furthermore, the conductive system includes at least four electrodes, and the electrode sensing ends are distributed at the two ends of the first area, the second area, and the third area, at least the first area or the second area or the third area is the reaction area, and the first pump body can drive the microfluidic sample to enter the first area, the second area, and the third area in sequence.

[0015] Furthermore, the microfluidic sample detection device also includes a starting electrode, whose sensing end is located upstream of the second valve body and together with the first electrode sensing end downstream thereof, forms a starting area in the microfluidic channel. In response to the microfluidic sample flowing into the starting area and contacting the starting signal generated by the first electrode sensing end, the first valve body is closed and the second valve body is opened.

[0016] Preferably, the first lock body is located in the microfluidic channel between the first electrode sensing end and the second electrode sensing end, and the first electrode sensing end and the second electrode sensing end are close to the second valve body and located downstream of the second valve body, thereby ensuring that the first area and the second area are downstream of the second valve body, ensuring that the first pump body realizes its driving function while maximizing the duration of the driving force of the first pump body.

[0017] When the microfluidic sample detection device includes at least one reaction zone, in particular, the reaction zone is pre-installed with magnetic particles combined with anti-erythrocyte antibodies, which are designed to specifically bind to erythrocytes in a blood sample.

[0018] When the microfluidic sample detection device comprises at least one reaction zone, in particular, the reaction zone is pre-installed with target antibodies labeled with magnetic particles and designed to specifically bind to the target in the sample.

[0019] When the microfluidic sample detection device includes at least one reaction zone, in particular, the reaction zone is pre-installed with antibodies labeled with fluorescent particles, and the antibodies are designed to specifically bind to the target in the sample.

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

[0021] Furthermore, the microfluid sample is specifically processed before combining with the antibodies in the two reaction zones. The present invention also provides a microfluid sample detection system, comprising a detection device and the above-mentioned microfluid sample detection device, the microfluid sample detection device is loaded in the detection device, and the detection device is electrically / signally connected to the electrodes of the microfluid detection device, so as to complete the opening or closing control of the first valve body and the second valve body, and the compression or release control of the first pump body through the actuator.

[0022] Furthermore, the microfluidic sample detection device is electrically connected to the detection device electrode through the electrical contact pin. The beneficial effects of the present invention include: 1. The microfluidic sample detection device has a simple structure and is easy to manufacture. 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 microfluidic sample detection device; 2. The liquid collection area in the microfluidic sample detection device is located at the far downstream of the microfluidic channel and will not pollute the detection equipment; 3. The valve body and the lock body in the microfluidic sample detection device cooperate to complete the sample quantification. After being applied to in vitro diagnostic electrochemical products, the accuracy and repeatability are better than traditional in vitro diagnostic electrochemical products. BRIEF DESCRIPTION OF THE DRAWINGS

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

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

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

[0026] Figure 1-4 A fourth implementation mode of the microfluidic detection device of the present invention.

[0027] Figure 2 A fifth implementation of the microfluidic detection device of the present invention.

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

[0029] Figure 3-2 Embodiment 7 of the microfluidic detection device of the present invention.

[0030] Figure 4-1 Embodiment 8 of the microfluidic detection device of the present invention.

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

[0032] Figure 5 A combined implementation mode of the microfluidic detection device of the present invention.

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

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

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

[0036] Fig. 9 Example 6 Results of a study on the accuracy of the product of the present invention.

[0037] Fig.10 Example 6A Results of a study on the accuracy of conventional products.

[0038] Fig.11 Example 6B Results of a study on the accuracy of conventional products. DETAILED DESCRIPTION

[0039] 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.

[0040] Example 1

[0041] like Figure 1-1 As shown, one embodiment of the microfluid detection device of the present invention is shown, which includes a fluid inlet 11, a fluid outlet 12, a microfluid channel 13, a liquid collection area 14, a reaction area 131, a pump body 41, a valve body, and a lock body 21. The fluid inlet 11 is used to add a microfluid sample to be tested, the fluid outlet 12 is communicated with 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 the same as the volume of the gas discharged from the fluid outlet at the same time.

[0042] The liquid receiving area 14 is downstream of the microfluidic channel and also downstream of the reaction area, and is used to receive the microfluidic 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 microfluidic 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 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.

[0043] 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 a portion of the microfluidic channel. The microfluidic sample to be tested is designed to react with a fixed preset reagent in the reaction zone during the flow in the microfluidic channel.

[0044] The pump body 41 is located upstream of the reaction zone and is used to passively drive the microfluidic sample to flow in the microfluidic channel. In other words, by controlling the pressure applied to the pump body, a driving force can be provided for the flow of the microfluidic sample, and the driving force drives the microfluidic sample to be tested that enters the microfluidic channel to continue to flow downstream. The pressure value applied to the pump body can be preset and associated with the travel position of the microfluidic sample in the microfluidic channel after entering the microfluidic channel. In other words, when the pressure value applied to the pump body is preset 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.

[0045] The valve body includes two valve bodies, one of which 31 is arranged downstream of the fluid inlet, and is used to control the connection between the fluid inlet and the microfluidic channel. It is in an open state before the microfluidic sample enters the microfluidic channel. The second valve body 32 is arranged downstream of the first valve body and is located between the pump body and the microfluidic channel. It is used to control the connection between the pump body and the microfluidic channel. In other words, the second valve body controls the first pump body to determine whether to provide driving force for the microfluidic sample. The second valve body is also used to assist in determining the volume of the microfluidic sample that ultimately participates in the reaction. The working states of the two valve bodies are opposite, that is, when one valve body is opened, the second valve body is closed, and when the second valve body is opened, the first valve body is closed. When the second valve body is opened and the first 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.

[0046] 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.

[0047] 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.

[0048] The lock body 21 is arranged downstream of the second valve body, and is used to limit the volume of the microfluid sample to be tested. When the microfluid sample to be tested enters the microfluid channel from the fluid inlet and the front end of the microfluid sample to be tested contacts the lock body, the flow stops. The volume of the microfluid sample to be tested between the fluid inlet and the lock body in the microfluid channel is the limited volume of the microfluid sample to be tested. In a further embodiment, the lock body and the second valve body cooperate to determine the volume of the microfluid sample to be tested. When the microfluid sample to be tested enters the microfluid channel from the fluid inlet and the front end of the microfluid sample to be tested contacts the lock body, the flow stops. After closing one of the valve bodies, the second valve body is opened, and the pump body drives the microfluid sample to be tested between the second valve body and the lock body to flow downstream in the microfluid channel. The part of the microfluid sample flowing downstream is the limited volume of the microfluid sample to be tested.

[0049] like Figure 6 As shown, it is a schematic diagram of the structure of the detection device loaded with a microfluidic detection device, which includes a main control board, a display screen, a load position, a magnetic module, a temperature module, an optical module, an actuator, a battery, a power interface, and a timer in order to realize the detection function of the photochemical method. Of course, if only a certain detection is set, the detection device can also be simplified, for example, only a main control board, a load position, an actuator, an optical module, a power interface, and a timer.

[0050] 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.

[0051] 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 then 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.

[0052] 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.

[0053] At least one reaction waiting time countdown is pre-set in the timer to determine a fixed time period for the reaction of the microfluid sample in the microfluid channel. In another embodiment, a countdown for the waiting time for adding the microfluid sample can also be set, and the time when the detection device and the detection device 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 microfluid sample to be tested has been successfully added. When the timer is used to determine that the sample addition is successful, it is not necessary to use the above-mentioned light detection unit to determine that the sample addition is successful. In another embodiment, a countdown for the time when the microfluid sample reaches a fixed area (such as a reaction area) can also be set, and the countdown is performed from the time when the microfluid sample starts to flow. When the countdown ends, the microfluid sample reaches the fixed area. The actuator provides pressure to the pump body of the detection device, and provides pressure to the pump body after the microfluid sample to be tested is successfully added, and compresses the pump body to drive the sample to flow to the downstream area of ​​the microfluid channel in the detection device. The downstream area here refers to, for example, a processing area, a reaction area, a detection area, etc. In some embodiments, the pressure value applied by the actuator to the pump body can be preset and associated with the position of the microfluidic sample in the microfluidic channel after it enters the microfluidic channel. That is, when the pressure value applied to the pump body is preset 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 countdown of the time when the microfluidic sample reaches a fixed area may not be set in the above timer. The actuator provides pressure to the valve body to close the valve body to close the microfluidic channel, or releases pressure to open the valve body to allow the microfluidic channel to pass through. 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.

[0054] 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.

[0055] The system consisting of the above-mentioned microfluidic sample detection device and detection equipment can be used as a microfluidic sample extraction system for separation of red blood cells and plasma in blood samples and extraction of plasma. Red blood cells are used as targets, and the reagents preset in the reaction area are magnetic particles combined with anti-red blood cell antibodies.

[0056] 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 and close the second valve body. The first countdown of the timer starts, and at the same time, the blood sample is added to the fluid inlet and enters the microfluidic channel. During the process of the blood sample entering the microfluidic channel, 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 entering. The front end of the blood sample contacts the lock body and stops entering.

[0057] After the first countdown of the timer ends, the main control board determines that the sample addition has been successful, and instructs the actuator to close one of the valve bodies, open the second valve body, and compress the pump body to drive the blood sample from the second valve body to the lock body into the reaction area. The second countdown of the timer begins. During 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 entering.

[0058] The front end of the blood sample contacts the end end of the reaction zone, that is, the blood 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, and the third countdown of the timer begins. The blood sample reacts with the anti-red blood cell antibodies combined with the magnetic particles in the reaction zone, and the red blood cells in the blood sample combine with the antibodies to form a magnetic particle-anti-red blood cell antibody-red blood cell complex. The third countdown of the timer ends, and the main control board commands the magnetic shielding body in the magnetic module to move so as to leave the area between the magnet and the microfluidic channel, so that the reaction zone is exposed to the magnetic field generated by the magnet. The red blood cells combined with the antibodies are adsorbed on the microfluidic channel wall of the reaction zone due to the action 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. Finally, the plasma that has lost the red blood cells can flow out of the fluid outlet for subsequent use.

[0059] 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.

[0060] Example 2

[0061] The microfluidic sample detection system composed of the microfluidic sample detection device and the detection equipment in Example 1 is applied to the detection of urine sugar, with glucose as the target. The reagent pre-installed in the reaction area 131 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.

[0062] Turn on the power of the detection equipment, 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. The microfluidic detection device is connected to the microfluidic detection device signal. The main control board commands the actuator to open one of the valve bodies and close the second valve body. The first countdown of the timer starts. At the same time, the urine sample to be tested is added to the fluid inlet and enters the microfluidic channel. During the process of the urine sample to be tested entering the microfluidic channel, part of the gas is discharged through the fluid outlet. The volume of the discharged gas is proportional to the volume of the urine sample to be tested. The front end of the urine sample to be tested contacts the lock body, and the urine sample to be tested stops entering. The first countdown of the timer ends, and the main control board receives a signal that the countdown ends.

[0063] In another embodiment, the load position does not generate a loaded signal, but transmits a loaded signal to the main control board by manually clicking a button on the detection device.

[0064] The main control board determines that the sample addition has been successful, and instructs the actuator to close one of the valve bodies, open the second valve body, compress the pump body to drive the second valve body to the lock body so that the urine sample to be tested enters the reaction area, and the second countdown of the timer begins. During the process of the urine 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 urine sample to be tested that enters.

[0065] The front end of the urine sample to be tested contacts the end 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, and 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 glucose exists in the urine sample and commands the display screen to display the test 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 of the detection equipment and discarded.

[0066] In another embodiment, there is no first countdown, but successful sample addition is determined by observation, and a cover body that can generate a successful sample 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 front end of the urine sample to be tested contacts the lock body and stops flowing, the cover body is closed, and the cover body generates a successful sample addition signal and transmits it to the main control board to determine that the sample addition is successful; in another embodiment, there is no first countdown, and successful sample addition of the urine sample is determined by the optical module; in another embodiment, there is no second countdown, and the pressure value applied by the actuator to the pump body is used to determine that the urine sample has filled the reaction area.

[0067] Example 3

[0068] like Figure 1-2 As shown, it is the second embodiment of the microfluidic detection device of the present invention, based on Example 1, it also includes a conductive system composed of electrodes. The conductive system includes two electrodes, electrode one 51 and electrode two 52, wherein electrode one also includes electrode contact one 511 and electrode sensing end one 512, electrode two also includes electrode contact two 521 and electrode sensing end two 522, and electrode sensing end one 512 and electrode sensing end two 522 are respectively located at the two ends of the reaction zone 131, that is, electrode sensing end one is located at the starting end of the reaction zone, and electrode sensing end two is located at the ending end of the reaction zone, and electrode sensing end one and electrode sensing end two are both located downstream of valve body two and close to valve body two, thereby ensuring that the reaction zone is downstream of valve body two to ensure that the pump body realizes its driving function. The lock body can be located in the reaction zone or outside the reaction zone, but is located upstream of the ending end of the reaction zone. As shown Figure 1-2 As shown in (a), the lock body is in the reaction zone and is located downstream of the starting end of the reaction zone, that is, the lock body is located in the microfluidic channel between the electrode sensing end 1 and the electrode sensing end 2, and the electrode sensing end 1 and the electrode sensing end 2 are close to the valve body 2 and are also located downstream of the valve body 2. Figure 1-2 As shown in (b), the lock body is outside the reaction zone and close to the electrode sensing end 1, and is located in the microfluidic channel upstream of the electrode sensing end 1, that is, the lock body is not between the electrode sensing end 1 and the electrode sensing end 2, but is located downstream of the valve body 2.

[0069] like Figure 7 As shown, it is a schematic diagram of the structure of the detection equipment of the load microfluid detection device, in order to realize the detection function including the electrochemical method, it includes a main control board, a display screen, a load position, a magnetic module, a temperature module, an electrical module, an actuator, a battery, a power interface, and a timer. Of course, if it is set to perform only a certain detection, the detection equipment can also be simplified, for example, only a main control board, a load position, an actuator, an electrical module, a power interface, and a timer.

[0070] The power interface provides power for the instrument 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 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.

[0071] 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.

[0072] 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.

[0073] In some embodiments, a countdown for the waiting time for adding microfluid samples can be set in a timer, and the time when the detection device and the detection equipment complete the electrical connection of the electrodes is used as the starting point for the waiting time for adding samples. When the time in the timer is passed, it is determined that the microfluid sample to be tested has been added successfully.

[0074] The actuator provides pressure to the valve body to control the opening and closing of the valve body. The actuator provides pressure to the pump body of the detection device. After the microfluid sample to be tested is successfully added, the pump body is compressed to drive the sample to flow to the downstream area of ​​the microfluidic channel in the detection device. The downstream area here refers to, for example, a processing area, a reaction area, a detection area, etc. In some embodiments, the pressure value applied by the actuator to the pump body can be preset and associated with the travel position of the microfluidic sample in the microfluidic channel after entering it. That is, when the pressure value applied to the pump body is preset 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. In such an embodiment, there is no need to use a timer to determine that the addition is successful and that the microfluidic sample has reached a fixed area.

[0075] 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.

[0076] 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.

[0077] The microfluidic sample detection system composed of the above-mentioned microfluidic sample detection device and detection equipment can also be used for separation of red blood cells and plasma in blood samples and extraction of plasma. The reagent preset in the reaction area is magnetic particles combined with anti-red blood cell antibodies.

[0078] Compared with Example 1, the difference is that after turning on the power of the detection equipment, the main control board commands the electrical module to apply a weak electrical signal to the two electrical contact pins corresponding to electrode one and electrode two. After the blood sample enters the microfluidic channel, the front end of the blood sample contacts the electrode sensing end two, 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 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 contact pins corresponding to electrode contact one and electrode contact two, which can replace the countdown function of a timer.

[0079] Example 4

[0080] The microfluidic sample detection system composed of the microfluidic sample detection device and the detection equipment in Example 3 is applied to the detection of blood glucose, with glucose as the target. The reagent preset in the reaction area 131 is a preparation formed by glucose dehydrogenase, an electron mediator, a surfactant, and a buffer solution.

[0081] 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, 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 the loaded signal to the main control board by manually clicking a button on the detection device.

[0082] The fluid inlet is exposed to the environment, and the main control board receives a signal from the electrical connection of the electrodes, and commands the actuator to open one of the valve bodies and close the second valve body. The first countdown of the timer starts, and the main control board commands the electrical module to apply a weak electrical signal between the two electrical contacts corresponding to electrode one and electrode two. At the same time, the blood sample to be tested is added to the fluid inlet and enters the microfluidic channel. During the process of the blood sample to be tested entering the microfluidic channel, 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. The front end of the blood sample to be tested contacts the lock body, and the blood sample to be tested stops entering.

[0083] After the first countdown of the timer ends, the main control board determines that the sample addition has been successful, and instructs the actuator to close one of the valve bodies, open the second valve body, and compress the pump body to drive the blood sample to be tested from the second valve body to the lock body into the reaction area. During 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.

[0084] 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 react with the preset reagent in the reaction zone and electron transfer to occur. The current representing the reaction result signal is sensed by the electrode. The electrons are received at terminal one and electrode sensing terminal two, and transmitted to the detection device through electrode contact one, electrode contact two, electrical contact pin one, and electrode contact pin two. The main control board receives and detects the current generated by the electron transfer, processes and determines the glucose concentration 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, and the used microfluidic sample detection device is taken out from the detection device and discarded.

[0085] Example 5

[0086] like Figure 1-3 As shown, the third embodiment of the microfluidic detection device of the present invention is based on Example 3 or any other embodiment (such as Example 1), 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.

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

[0088] The difference from other embodiments is that after turning on the power of the detection device, the main control board 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 to the electrode sensing end three and the electrode sensing end four in the microfluidic channel branch, so that after a loop is formed between the electrode three and the electrode four, it can be determined 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.

[0089] Example 6

[0090] like Figure 2As shown, it is the fifth embodiment of the microfluid detection device of the present invention. On the basis of Example 3, 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. 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. Electrode sensing end one is located at the starting end of region one, electrode sensing end two is located at the ending end of region one and is also the starting end of region two, electrode sensing end five is located at the ending end of region two, electrode sensing end one, electrode sensing end two and electrode sensing end five are all located downstream of valve body two, and electrode sensing end one is close to valve body two, so as to ensure that region one and region two are downstream of valve body two, while ensuring that the pump body realizes its driving function, the duration of the pump body driving force is maximized. The lock body is in area one and downstream of the starting end of area one, that is, the lock body is between electrode sensing end one and electrode sensing end two and downstream of valve body two; or the lock body is outside area one and upstream of the starting end of area one, that is, the lock body is outside electrode sensing end one and electrode sensing end two and downstream of valve body two.

[0091] For example, red blood cells are used as the first target, and the reagents pre-set in area 1 are magnetic particles combined with anti-red blood cell antibodies. Glucose is used as the second target, and the reagents pre-set in area 2 are preparations containing glucose oxidoreductase and electron mediators. In this case, both area 1 and area 2 are reaction areas.

[0092] 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 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 and close the second valve body. The fluid inlet is exposed to the environment, and the first countdown of the timer starts. At the same time, the blood sample to be tested is added to the fluid inlet and enters the microfluidic channel. During the process of the blood sample to be tested entering the microfluidic channel, 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 entering. The front end of the blood sample to be tested contacts the lock body, and the blood sample to be tested stops entering.

[0093] 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, open valve body two, and compress the pump body to drive the blood sample to be tested from valve body two to the lock body into area one. During the process of the blood sample to be tested entering area one, 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 entering.

[0094] 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, and starts applying a weak electrical signal between the two electrical contact pins corresponding to the electrode contact 2 and the electrode contact 5. The second countdown of the timer starts, waiting for the blood sample to be tested to react with the anti-red blood cell antibody in area 1 with the magnetic particles. The red blood cells in the blood sample bind to 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 so as to leave the area 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 antibodies are adsorbed on the wall of the microfluidic channel in area one due to the action 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 flow from area one into area two. During this 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 that has lost the red blood cells and entered area two.

[0095] 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 glucose 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 glucose concentration in the plasma and commands the display screen to display the detection result. At the same time, it commands the actuator 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.

[0096] The above test was repeated 10 times with blood samples with glucose contents of 50 mg / dL, 100 mg / dL, 250 mg / dL, 400 mg / dL, and 550 mg / dL, and the test results were obtained. A biochemical standard instrument was used as a control. The same blood samples were tested 10 times on the existing excellent brand A traditional electrochemical and average brand B traditional electrochemical blood glucose detection products, and the test results were obtained, 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 glucose 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 glucose content in the blood sample measured by the present invention are closer to the true value and have higher accuracy than existing products with excellent performance.

[0097] Table 1 Comparative experimental results of Example 6

[0098]

[0099] Of course, if the electrodes do not exist, region 1 and region 2 can also be formed in the microfluidic channel, and the preset reagent in region 2 is replaced with anthrone reagent, which changes color when reacting with glucose, and the reaction result is detected through the optical module.

[0100] Example 7

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

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

[0103] The microfluidic detection device is formed by bonding a substrate and a covering film, a frame-shaped electrode is printed on the substrate, and an electrode frame-shaped 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-shaped area and the microfluidic channel in the substrate jointly form an area for gathering pre-set reagents, and the two sides of the electrode frame-shaped area parallel to each other in the horizontal direction can be used as the electrode sensing ends of the frame-shaped electrode.

[0104] Example 8

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

[0106] For example, C-reactive protein is used as the target. The reagent pre-installed in area 1 is a C-reactive protein antibody bound to magnetic particles, that is, a C-reactive protein antibody labeled with magnetic particles. The reagent pre-installed in area 2 is a 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.

[0107] 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 and close the second valve body. The fluid inlet is exposed to the environment, the first countdown of the timer starts, and at the same time, the blood sample to be tested is added to the fluid inlet and enters the microfluidic channel. The front end of the blood sample to be tested contacts the lock body, and the blood sample to be tested stops entering. 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 contact pins corresponding to the electrode 1 and the electrode 2, and instructs the actuator to close the valve body 1, open the valve body 2, and compress the pump body to drive the blood sample to be tested from the valve body 2 to the lock body into the area 1. The front end of the blood sample to be tested contacts the termination end of the 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 instructs the actuator to stop compressing the pump body, and at the same time instructs 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, and between the two electrical contact pins corresponding to the electrode contact 2 and the electrode contact 6. The second countdown of the timer begins, waiting for the blood sample to be tested to react with the magnetic particle-bound C-reactive protein antibody in the area 1, and the C-reactive protein in the blood sample to be tested binds to 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 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 two and electrode contact six. The main control board commands the actuator to stop compressing the pump body, and at the same time commands the electrical module to stop applying a weak electrical signal between the two electrical contact pins corresponding to electrode contact two and electrode contact six, and start applying a weak electrical signal between the two electrical contact pins corresponding to electrode contact six and electrode contact five. 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. A conductive path is formed between electrode sensing end two and electrode sensing end six through the blood sample containing the aforementioned complex, and an 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 main control board commands the magnetic shield in the magnetic module to move so as to leave the space between the magnet and the microfluidic channel, so that area three is exposed to the magnetic field generated by the magnet. Within the magnetic field, 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 action of the magnetic field, and 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, and 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.

[0108] 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.

[0109] 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.

[0110] Example 9

[0111] On the basis of Example 8, it also includes electrode seven, which includes electrode contact seven and electrode sensing end seven, forming region one, region two, region three and region four in the microfluidic channel. At this time, region three is the most downstream region.

[0112] That is, electrode sensing end 5 is located at both the end of region 3 and the start of region 4, and electrode sensing end 7 is located at the end of region 4. In this case, region 1 is the pretreatment region, region 2 and region 3 are the reaction regions, and region 4 is the detection region.

[0113] For example, 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.

[0114] 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, electrode contact six, and electrode contact seven, 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 and close the second valve body. The fluid inlet is exposed to the environment, the first countdown of the timer starts, and at the same time, the blood sample to be tested is added to the fluid inlet and enters the microfluidic channel. The front end of the blood sample to be tested contacts the lock body, and the blood sample to be tested stops entering.

[0115] 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 contact pins corresponding to the electrode 1 and the electrode 2, and instructs the actuator to close the valve body 1, open the valve body 2, and compress the pump body to drive the blood sample to be tested from the valve body 2 to the lock body into the area 1. The front end of the blood sample to be tested contacts the terminal end of the 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 instructs the actuator to stop compressing the pump body, and at the same time instructs 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 begins, waiting for the blood sample to be tested to be fully incubated and dissociated by the VD dissociation solution in the area 1. In order to fully release the bound VD in the blood sample to be tested, the main control board can also instruct the temperature module to heat the area 1 before the second countdown of the timer begins. 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 achieved 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 continue compressing 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.

[0116] 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.

[0117] 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.

[0118] Example 10

[0119] like Figure 4-1 As shown, the eighth embodiment of the microfluidic detection device of the present invention, in Example 1 or any other embodiment, includes 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 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 enclosed to form a starting area, the starting area is close to the microfluid inlet, electrode sensing end eight is the starting end of the starting area, electrode sensing end one is the ending end of the starting area, and the existence of the starting area can replace the function of the first countdown of the timer before the successful addition of the sample.

[0120] like Figure 4-2As shown, this is a ninth embodiment of the microfluidic detection device of the present invention. Figure 4-1 In another embodiment, the lock body includes an electrode 2, an electrode contact 2, and an electrode sensing end 2. The lock body is in the microfluidic channel between the electrode sensing end 1 and the electrode sensing end 2, and is located downstream of the electrode sensing end 1. The electrode sensing end 1 and the electrode sensing end 2 are close to the valve body 2 and are also located downstream of the valve body 2. In another embodiment, reference Figure 1-2 (b) is different from Figure 4-2 In the embodiment, the lock body is not in the microfluidic channel between the electrode sensing end 1 and the electrode sensing end 2, but in the microfluidic channel between the electrode sensing end 1 and the electrode sensing end 8. The lock body is located upstream of the electrode sensing end 1, and the electrode sensing end 1 and the electrode sensing end 2 are still close to the valve body 2 and are also located downstream of the valve body 2.

[0121] 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 actuator to open valve body one and close valve body two, while commanding 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 microfluidic sample contacts electrode sensing end one, a conductive path is formed between electrode sensing end one and electrode sensing end eight through the microfluidic sample, 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 loaded, and commands to close valve body one and open valve body two. The microfluidic sample in the microfluidic channel between valve body two and electrode sensing end one is the determined microfluidic sample volume, thereby serving as a double guarantee for confirming the successful loading together with the lock body.

[0122] Embodiment 11

[0123] The microfluidic detection device of the present invention can be combined on the basis of Examples 1 to 10 and applied to the combined detection of multiple indicators. Pregnant women are at risk of gestational diabetes and anemia, so glucose, beta-hydroxybutyric acid and hemoglobin in the blood should be monitored during pregnancy.

[0124] like Figure 5 As shown, a device for simultaneously detecting glucose, β-hydroxybutyrate and hemoglobin in blood, device unit one 101 is used to detect glucose, device unit two 102 is used to detect β-hydroxybutyrate, device unit three 104 and device unit four are used to detect hemoglobin 105.

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

[0126] 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.

[0127] 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.

[0128] 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, 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.

[0129] 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.

[0130] 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.

[0131] 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.

[0132] 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 mutually exclusive of other embodiments.

[0133] 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 microfluidic sample detection device, characterized in that: It includes a fluid inlet, a microfluidic channel and a fluid outlet, and: A reaction zone is provided between the fluid inlet and the fluid outlet and partially overlaps with the microfluidic channel; a liquid collection zone is provided downstream of the reaction zone, and the fluid outlet is the outlet of the liquid collection zone; A first pump body, disposed upstream of the reaction zone, and providing a driving force for the flow of the microfluidic sample when communicating with the microfluidic channel; A first valve body, disposed downstream of the fluid inlet and controlling the connection and disconnection between the microfluidic channel and the fluid inlet; A second valve body, which is disposed downstream of the first valve body and controls the connection and disconnection between the first pump body and the microfluidic channel, and the working states of the first valve body and the second valve body are opposite; The first lock body is disposed downstream of the second valve body and defines the volume of the microfluid sample together with the second valve body.

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

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

4. The microfluidic sample detection device according to claim 1, characterized in that: It also includes a conductive system composed of electrodes, wherein the conductive system includes at least two electrodes, and the electrode sensing ends are respectively located at two ends of the reaction zone.

5. A microfluidic sample detection device according to claim 4, characterized in that: The conductive system includes at least three electrodes, and the electrode sensing ends are distributed at both ends of the first area and the second area. At least the first area or the second area is a reaction area, and the first pump body can drive the microfluid sample to enter the first area and the second area in sequence.

6. A microfluidic sample detection device according to claim 5, characterized in that: The conductive system includes at least four electrodes, and the electrode sensing ends are distributed at both ends of the first area, the second area and the third area. At least the first area or the second area or the third area is a reaction area. The first pump body can drive the microfluid sample to enter the first area, the second area and the third area in sequence.

7. A microfluidic sample detection device according to any one of claims 4 to 6, characterized in that: It also includes a starting electrode, wherein the starting electrode sensing end is located upstream of the second valve body and together with the first electrode sensing end downstream of the second valve body, forms a starting area in the microfluidic channel. In response to the microfluidic sample flowing into the starting area and contacting the starting signal generated by the first electrode sensing end, the first valve body is closed and the second valve body is opened.

8. A microfluidic sample detection device according to any one of claims 4 to 6, characterized in that: The first lock body is located in the microfluidic channel between the first electrode sensing end and the second electrode sensing end. The first electrode sensing end and the second electrode sensing end are close to the second valve body and are located downstream of the second valve body.

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

10. A microfluidic sample detection device according to claim 1 or 2 or 4 or 5, characterized in that: The reaction area is pre-installed with magnetic particles combined with anti-erythrocyte antibodies, and the antibodies are designed to specifically bind to erythrocytes in a blood sample.

11. A microfluidic sample detection device according to any one of claims 1 to 6, characterized in that: The reaction area is pre-installed with target antibodies labeled with magnetic particles, and the antibodies are designed to specifically bind to the target in the sample.

12. A microfluidic sample detection device according to any one of claims 1 to 6, characterized in that: The reaction area is pre-installed with antibodies labeled with fluorescent particles, and the antibodies are designed to specifically bind to the target in the sample.

13. A microfluidic sample detection device according to claim 2 or 3 or 5 or 6, characterized in that: The method comprises two reaction zones, wherein a first antibody of a target object labeled with magnetic particles and a second antibody labeled with fluorescent particles are respectively pre-installed in the two reaction zones. The first antibody and the second antibody are designed to specifically bind to the target object in the sample. After the target object in the microfluidic sample binds to the first antibody and the second antibody, a double antibody sandwich complex is formed.

14. A microfluidic sample detection device according to claim 13, characterized in that: The microfluidic sample is specifically treated before being combined with the first antibody and the second antibody.

15. A microfluidic sample detection system, comprising a detection device and a microfluidic sample detection apparatus as described in any one of claims 1 to 14, wherein the microfluidic sample detection apparatus is loaded in the detection device, and the detection device is electrically / signally connected to electrodes of the microfluidic detection apparatus, thereby completing the control of opening or closing of the first valve body and the second valve body, and the control of compression or release of the first pump body through an actuator.

16. A microfluidic sample detection system according to claim 15, characterized in that: The microfluid sample detection device is electrically connected to the detection equipment electrode via an electrical contact pin.

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