A split microfluidic chip and its working method

Abstract

The invention discloses a split microfluidic chip and a working method thereof; in the chip, a reagent storage module is connected to a reaction module through a connection module, and the reaction module is slidably arranged in a slide groove of the connection module; a plurality of solution containing cavities are arranged on the reagent storage module, a sealing film is attached to the top of the reaction module, and a plurality of solution flow cavities are arranged on the reaction module; when working, the reaction module and the reagent storage module abut against each other, and when the solution containing cavities are respectively connected to the atmosphere, negative pressure is drawn to respectively realize the directional flow and release of the solution in the solution containing cavity in the microfluidic chip to the solution flow cavity; the invention adopts an integrated structural design and is easy to operate, can realize sample sampling, cleaning, nucleic acid adsorption, elution and reaction in the microfluidic chip, and realizes the instant detection of pathogenic microorganisms.

Description

Technical Field

[0001] The present invention belongs to the technical field of microfluidic chips, and in particular, relates to a split microfluidic chip and a working method thereof. Background Art

[0002] Microfluidics refers to the manipulation of liquids at a submillimeter scale, where the submillimeter scale is generally a few microns to hundreds of microns. Microfluidics integrates the basic operating units involved in the biological and chemical fields, and even the functions of the entire laboratory, including sampling, dilution, reaction, separation, and detection, on a small chip, so microfluidic chips are also called chip laboratories (Lab-on-a-Chip). This chip is generally composed of various liquid reservoirs and interconnected microchannel networks, which can greatly shorten the sample processing time and achieve maximum utilization efficiency of reagents and consumables by precisely controlling the flow of liquids.

[0003] In recent years, the application of microfluidic chips in fully automated, integrated biological testing POCT (full name: point-of-care testing, Chinese name: instant testing) has emerged, which has broad application prospects and important application value.

[0004] The above-mentioned microfluidic chip is applied to fully automated and biologically integrated detection, which usually requires a closed reaction system, that is, the reaction system is isolated from the outside world, and various reagents need to be stored in isolation from the outside world in space, and released according to the process when needed to form contact and mixing to react. The existing technology usually adopts the design of valves and pipelines to achieve the addition of reaction materials in sealed body reactions, but the valve and pipeline method is very difficult in the assembly and design of microfluidic chip products, which leads to a high price of microfluidic chip products, which is not conducive to the promotion and application of POTC.

[0005] The existing microfluidic chips, valves and pipelines are very difficult to assemble and design in microfluidic chip products, resulting in high prices for microfluidic chip products, which is not conducive to the promotion and application of microfluidic chip instant detection and has high cost of use. Summary of the invention

[0006] In view of the deficiencies of the above-mentioned prior art, the present invention provides a split microfluidic chip and a working method thereof, which has a reasonable structure, low cost, non-interference and non-interconnection of the functional areas, and easy operation, thereby solving the technical problems of solution storage and transfer of the microfluidic chip in the above-mentioned prior art.

[0007] The technical solution of the present invention is specifically described as follows.

[0008] The present invention provides a split microfluidic chip, which includes a reagent storage module, a reaction module and a connection module; the reagent storage module is connected to the reaction module through the connection module, the connection module is provided with a slide groove, the reaction module is slidably arranged in the slide groove, and when working, the reaction module is pushed to achieve mutual abutment between the reaction module and the connection module; wherein:

[0009] The reagent storage module is provided with a plurality of solution containing chambers, the top openings of the solution containing chambers are isolated from the outside by a sealing film, the bottom openings of some of the solution containing chambers are connected to the first spike portion, the first spike portion is open at both ends and is hollow, the bottom of another part of the solution containing chambers is sealed with a sealing film, a connecting channel is provided near the bottom, the top of the connecting channel is a spike portion containing channel, a second spike portion is slidably provided in the spike portion containing channel, the second spike portion is bounded by a through hole, the upper part is solid, the lower part is hollow and a through hole is provided on the side wall, when an external force is applied, the second spike portion slides downward in the spike portion containing channel so that the through hole on the second spike portion and the connecting channel provided near the bottom of the solution containing chamber are connected to each other;

[0010] A plurality of solution flow chambers are arranged on the reaction module, and a sealing film is arranged on the top opening of the solution flow chamber; the number and position of the solution flow chambers correspond to the solution containing chambers in the reagent storage module; when working, the first spike portion and the second spike portion pierce the sealing film at the top of the solution flow chamber, and under the action of negative pressure, the solution in the solution containing chamber is driven to flow to the corresponding solution flow chamber, thereby realizing the directional flow of the solution.

[0011] In the present invention, a first buckle and a second buckle are provided on the connecting module, a first limiting groove and a second limiting groove are provided on the side wall of the reaction module, a buckle connection is achieved between the reagent storage module and the connecting module through the first buckle, and a fixed connection is achieved between the reaction module and the connecting module in an initial state and a working state respectively through the cooperation between the second buckle and the first limiting groove and the second limiting groove.

[0012] In the present invention, on the reagent storage module, a third spike portion is used to apply an external force to a second spike portion arranged in the spike portion accommodating channel so that the second spike portion slides downward. The third spike portion has an opening at the top, a hollow structure at the top and a through hole on the side wall, and the sharp portion at the bottom is a closed structure.

[0013] In the present invention, the bottom of the first spike portion is sealed by an elastic rubber cap.

[0014] In the present invention, a connecting channel is arranged on the upper part of the solution containing chamber with the spike portion accommodating channel arranged inside, and the connecting channel, the spike portion accommodating channel and the solution containing chamber are all connected, so that during operation, the solution containing chamber and the atmosphere are connected through the connection between the through hole on the third spike portion and the connecting channel.

[0015] In the present invention, a premixing chamber and a reaction chamber are further provided on the reaction module, the solution flow chamber is interconnected with the premixing chamber through the flow channel, and the premixing chamber is connected with the reaction chamber through the flow channel.

[0016] In the present invention, the solution containing chamber is used to store sampling liquid, cleaning liquid, elution liquid and reaction liquid respectively; a float is arranged in the solution containing chamber for storing the sampling liquid, and a filter membrane is arranged in the solution flow chamber connected to the solution containing chamber for storing the sampling liquid to filter large particles of impurities in the sampling liquid; the flow channel connecting the solution flow chamber and the premixing chamber is a nucleic acid adsorption flow channel, in which a filler for adsorbing nucleic acids is arranged; there are a plurality of reaction chambers connected in series, and each reaction chamber has the same volume.

[0017] In the present invention, the solution containing chamber for filling and storing the reaction liquid includes a plurality of reaction liquid containing chambers interconnected by S-bends, and a paraffin block is arranged inside the S-bend to isolate and separate different reaction liquids; a side plate is arranged inside the reaction liquid containing chamber to receive the cooled paraffin block.

[0018] In the present invention, a first waste liquid chamber and a second waste liquid chamber are respectively provided on the reagent storage module and the reaction module. The first waste liquid chamber is used to collect excess sampling liquid and cleaning liquid through an external pipe, and the second waste liquid chamber is used to collect excess waste liquid flowing from the premixing chamber to the reaction chamber.

[0019] In the present invention, negative pressure ports are respectively provided on the reagent storage module and the reaction module, and the negative pressure ports are respectively connected to the first waste liquid chamber, the premixing chamber, and the second waste liquid chamber through external pipes and flow channels to realize the directional flow of the solution in the chip, thereby realizing sample sampling, cleaning, nucleic acid adsorption, elution and reaction mixing reactions.

[0020] The present invention also provides a working method of the above-mentioned split microfluidic chip, comprising the following two solution directional flow steps:

[0021] (1) External force pushes the reagent storage module to abut the reaction module and the reagent storage module, and the first spike portion connected to the bottom of the solution accommodating chamber pierces the sealing film at the top of the corresponding solution flow chamber, thereby realizing the mutual communication between the solution accommodating chamber and the solution flow chamber. At the same time, through the communication between the top of the solution accommodating chamber and the atmosphere, under the action of negative pressure, the solution in the solution accommodating chamber is driven to flow to the corresponding solution flow chamber;

[0022] (2) An external force drives the third spike portion to slide downward, respectively driving the second spike portion in the spike portion accommodation channel to slide downward to pierce the sealing film at the bottom of the corresponding solution accommodation cavity and the sealing film at the top of the flow-through cavity. Through the connection of the through holes on the second spike portion and the communication channel arranged near the bottom in the accommodation cavity, the mutual connection between the solution accommodation cavity and the solution flow-through cavity is realized. At the same time, through the connection of the through hole of the third spike portion in the upper part of the solution accommodation cavity and the communication channel in the upper part of the solution accommodation cavity, the connection between the solution accommodation cavity and the atmosphere is realized. Under the action of negative pressure, the solution in the solution accommodation cavity is driven to flow into the corresponding solution flow-through cavity.

[0023] Further, the working method of the above-mentioned split microfluidic chip provided by the present invention includes the following steps:

[0024] Step 1: An external force pushes the reagent storage module to abut against the reaction module, and at this time, the solution accommodation cavity for storing the sampling liquid, the solution accommodation cavity for storing the reaction liquid, and the corresponding solution flow-through cavity are respectively connected; at the same time, the sealing film at the top of the accommodation cavity for storing the sampling liquid is connected to the atmosphere. Under the action of negative pressure, the sampling liquid sequentially flows through the corresponding solution flow-through cavity, the nucleic acid adsorption flow channel, and the first waste liquid cavity; after the sampling liquid is pumped out, the floating ball drops to the bottom of the accommodation cavity to block the spike portion, and the first spike portion is not connected to the external atmosphere;

[0025] Step 2: An external force drives the third spike portion to slide downward, driving the second spike portion in the solution accommodation cavity for storing the cleaning liquid to slide downward to pierce the sealing film at the top of the corresponding solution flow-through cavity. The through hole on the second spike portion and the communication channel arranged near the bottom in the solution accommodation cavity are connected. At this time, the solution accommodation cavity for storing the cleaning liquid is connected to the atmosphere through the through hole on the upper third spike portion. Under the action of negative pressure, the cleaning liquid sequentially enters the corresponding solution flow-through cavity, the nucleic acid adsorption flow channel, and the first waste liquid cavity; after the cleaning liquid is pumped out, the external force continues to drive the second spike portion to slide downward so that the solution accommodation cavity is not connected to the external atmosphere;

[0026] Step 3: An external force drives the third spike portion to slide downward, driving the second spike portion in the solution accommodation cavity for storing the elution liquid to slide downward to pierce the sealing film at the top of the corresponding solution flow-through cavity. The through hole on the second spike portion and the communication channel arranged near the bottom in the solution accommodation cavity are connected. At this time, the solution accommodation cavity for storing the elution liquid is connected to the atmosphere through the through hole of the upper third spike portion. Under the action of negative pressure, the cleaning liquid enters the corresponding flow-through cavity, the nucleic acid adsorption flow channel, and the premixing cavity; after the cleaning liquid is pumped out, the external force continues to drive the second spike portion to slide downward so that the accommodation cavity is not connected to the external atmosphere;

[0027] Step 4: Connect the solution accommodation cavity for storing the reaction liquid to the atmosphere. Under the action of negative pressure, the reaction liquid flows into the premixing cavity;

[0028] Step 5: The solution in the premixing chamber flows into the reaction chamber under the action of negative pressure driving force, and the excess liquid enters the second waste liquid chamber.

[0029] Compared with the prior art, the present invention has the following beneficial effects:

[0030] The present invention realizes the sealed storage of sampling liquid, reaction liquid, eluent and cleaning liquid separately in the chip, and correspondingly releases accurate solutions in appropriate steps. The integrated structural design and simple operation realize the real-time detection of microfluidic chip experiments.

[0031] Moreover, through reasonable structural design of each flow channel and chamber, there is no problem of liquid cross-flow. The flow of various liquids in the chip is well controllable and they are not easy to contaminate each other.

[0032] Multiple waste liquid chambers are set in the chip. The waste liquid generated during the reaction enters the waste liquid chamber. After the reaction is completed, the chip can be discarded without generating aerosol pollution, thus protecting the laboratory environment from pollution. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 It is a structural exploded diagram of the split microfluidic chip of the present invention.

[0034] Figure 2 It is a rear view of the split microfluidic chip of the present invention.

[0035] Figure 3 This is a diagram of the initial state of the split microfluidic chip of the present invention.

[0036] Figure 4 It is a diagram of the abutment state of the split microfluidic chip of the present invention.

[0037] Figure 5 It is a state diagram of pumping cleaning fluid from the split microfluidic chip of the present invention.

[0038] Figure 6 This is a state diagram of the split microfluidic chip of the present invention after the cleaning liquid has been pumped out.

[0039] Figure 7 It is a state diagram of extracting reaction liquid of the split microfluidic chip of the present invention.

[0040] Figure 8 This is a state diagram of the split microfluidic chip of the present invention after the reaction liquid has been extracted.

[0041] Fig. 9 It is a structural diagram of the split microfluidic chip with a float of the present invention.

[0042] Numbers in the figure: 1-reagent storage module, 101-sampling liquid accommodating chamber, 102-cleaning liquid accommodating chamber, 103-eluting liquid accommodating chamber, 104-reaction liquid storage component, 1041-first reaction liquid accommodating chamber, 1042-S bend, 1043-second reaction liquid accommodating chamber, 105-floating ball, 106-first spike portion, 108-second spike portion, 110-third spike portion, 112-first through hole, 113-second through hole, 114-third through hole, 115-first connecting channel, 116-second connecting channel, 117-third connecting channel, 118-fourth connecting channel, 119-first waste liquid chamber , 122-waste liquid connection port, 123-side plate; 2-reaction module, 201-first limiting groove, 202-second limiting groove, 203-sampling liquid flow chamber, 204-cleaning liquid flow chamber, 205-elution liquid flow chamber, 206-reaction liquid flow chamber, 207-nucleic acid adsorption flow channel, 208-premixing chamber, 209-reaction chamber, 210-air vent, 211-waste liquid port, 212-premixing negative pressure port, 213-cleaning liquid suction port, 214-injection negative pressure port, 215-second waste liquid chamber, 216-first pair of interfaces, 217-second pair of interfaces, 218-third pair of interfaces, 219-fourth pair of interfaces;

[0043] 3-connecting module, 301-first buckle, 302-second buckle, 303-slide groove. DETAILED DESCRIPTION

[0044] The technical solution of the present invention is described in detail below in conjunction with the accompanying drawings and embodiments.

[0045] Please refer to Figure 1-Figure 9 The split microfluidic chip includes: a reagent storage module 1 (a sealing film is attached to the top surface of the reagent storage module 1), a reaction module 2 and a connection module 3. The reagent storage module 1 and the reaction module 2 are connected through the connection module 3. The connection module 3 is provided with a first buckle 301 and a second buckle 302. The reagent storage module 1 is fixed to the connection module 3 through the first buckle 301. The connection module 3 is provided with a slide groove 303. The reaction module 2 is slidably arranged in the slide groove 303. The reaction module 2 is also provided with a first limiting groove 201 and a second limiting groove 202 that are engaged with the second buckle 302. In the initial state, the second buckle 302 is engaged with the first limiting groove 201. When a reaction experiment is required, an external force pushes the reaction module 2 to slide in the slide groove 303 until the second buckle 302 is engaged with the second limiting groove 202. At this time, the reagent storage module 1 is in contact with the reaction module 2.

[0046] In an embodiment of the present invention, the split microfluidic chip is used to realize nucleic acid detection of pathogenic microorganisms.

[0047] The reagent storage module 1 is provided with: a sampling liquid accommodating chamber 101, a cleaning liquid accommodating chamber 102, an eluent accommodating chamber 103 and a reaction liquid storage component 104; a float 105 is provided in the sampling liquid accommodating chamber 101, and when the sampling liquid accommodating chamber 101 is filled with liquid, the float 105 floats.

[0048] The cleaning liquid accommodating chamber 102 and the eluting liquid accommodating chamber 103 on the reagent storage module 1 are respectively provided with a second spike portion accommodating channel, and the second spike portion 108 is respectively slidably arranged in the second spike portion accommodating channel; and the second spike portion 108 and the second spike portion channel are closely matched, and external force is applied to the third spike portion 110 respectively to make the second spike portion 108 slide in the second spike portion accommodating channel, and the third spike portion 110 is a hollow structure, the top opening forms a cavity, and the spike end is a closed structure. That is, the needle includes a sharp portion and a side wall formed along the sharp portion. Among them, the sharp portion constitutes a sealing structure, the side wall extends upward and forms an opening at the top, the opening is connected with the inside of the side wall to form a cavity together, and the side wall is provided with a third through hole 114.

[0049] The second spike portion 108 has a solid structure and a hollow structure. The hollow sections of the second spike portion 108 in the cleaning liquid accommodating chamber 102 and the eluting liquid accommodating chamber 103 are respectively provided with a first through hole 112 and a second through hole 113; the two second spike portions 108 are respectively divided by the first through hole 112 and the second through hole 113. The portion above the first through hole 112 and the second through hole 113 is a solid structure, and the portion below the first through hole 112 and the second through hole 113, including the first through hole 112 and the second through hole 113, is a hollow structure.

[0050] A sealing film (silicone film or pressure-sensitive film) is attached to the docking surface of the reaction module 2 and the reagent storage module 1. When an external force pushes the reaction module 2, the reaction module 2 and the reagent storage module 1 are abutted against each other. The first sharp thorn portion 106 below the sampling liquid accommodating cavity 101 pierces the sealing film at the first docking interface 216, and the first sharp thorn portion 106 below the reaction liquid storage component 104 pierces the sealing film at the fourth docking interface 219.

[0051] A first connecting channel 115 is provided in the cleaning liquid accommodating chamber 102 of the reagent storage module 1, and a second connecting channel 116 is provided in the eluent accommodating chamber 103 of the reagent storage module 1. The first and second connecting channels 115 and 116 are vertically arranged above the sealing membrane arranged at the bottom of the corresponding accommodating chamber, and the two third spike portions 110 are driven by external force to slide downward, respectively driving the second spike portions 108 in the cleaning liquid accommodating chamber 102 and the eluent accommodating chamber 103 to slide downward, respectively piercing the sealing membranes at the second docking interface 217 and the third docking interface 218, until the third through holes 114 of the two third spike portions 110 slide respectively. Move to the third communicating channel 117 (at this time, the first through hole 112 of the second spike portion 108 in the cleaning liquid accommodating chamber 102 is just at the first communicating channel 115), and the fourth communicating channel 118 (at this time, the second through hole 113 of the second spike portion 108 in the eluent accommodating chamber 103 is just at the second communicating channel 116). The third and fourth communicating channels 117 and 118 are horizontally arranged at the upper parts of the cleaning liquid accommodating chamber 102 and the eluent accommodating chamber 103, respectively. The third through hole 114 is connected to the third communicating channel 117 and the fourth communicating channel 118. The cleaning liquid accommodating chamber 102 and the eluent accommodating chamber 103 are connected through The hollow structure of the third spike portion 110 and the third through hole 114 are connected to the outside atmosphere; the third connecting channel 117 and the third spike portion accommodating channel and the cleaning liquid accommodating chamber 102 are all connected; the fourth connecting channel 118 and the fourth spike portion accommodating channel and the eluting liquid accommodating chamber 103 are all connected; until the first through hole 112 slides to the first connecting channel 115, the cleaning liquid flows into the hollow structure from the first through hole 112 and then enters the cleaning liquid flow chamber 204. After the cleaning liquid is exhausted, the external force continues to drive the third spike portion 110 to slide downward until the first through hole 112 slides downward away from the first connecting channel 115. The third spike portion 110 is driven to leave the second spike portion accommodating channel. At this time, the cleaning liquid accommodating chamber 102 is not connected with the outside world (nor with the reaction module 2); the second through hole 113 slides to the second connecting channel 116, and the eluent flows into the hollow structure from the second through hole 113 and then enters the eluent flow chamber 205. After the eluent is extracted, the external force continues to drive the third spike portion 110 to slide downward until the second through hole 113 slides downward away from the second connecting channel 116, driving the third spike portion 110 to leave the second spike portion accommodating channel. At this time, the eluent accommodating chamber 103 is not connected with the outside world (nor with the reaction module 2).

[0052] The reaction liquid usually contains multiple components. Compared with storing different solutions together, storing different components separately can store them separately for a longer time. For this reason, the structure of the reaction liquid storage component 104 is designed as follows: the reaction liquid storage component 104 is divided into three sections, the first reaction liquid accommodating chamber 1041, the S-bend 1042 and the second reaction liquid accommodating chamber 1043. The S-bend 1042 is provided with a paraffin block to separate the reaction liquids in the first reaction liquid accommodating chamber 1041 and the second reaction liquid accommodating chamber 1042. Separating different reaction liquids can make different reaction liquids be stored for a longer time without deterioration, thereby ensuring the accuracy of the experimental results. During the experiment, the microfluidic chip is heated, the paraffin melts, drips on the side plate, solidifies into a solid, floats on the surface of the solution and separates from the solution, so that the mixed reaction liquid smoothly enters the premixing chamber 208, the first reaction liquid accommodating chamber 1041, the S-bend 1042 and the second reaction liquid accommodating chamber 1043 are connected, and different reaction liquid combinations are mixed.

[0053] The reaction module 2 is provided with a sampling liquid flow chamber 203, a cleaning liquid flow chamber 204, an elution liquid flow chamber 205, a reaction liquid flow chamber 206, a nucleic acid adsorption flow channel 207, a premixing chamber 208, a reaction chamber 209, an air vent 210, a waste liquid port 211, and a premixing negative pressure port 212. A filter membrane is provided in the sampling liquid flow chamber 203 to filter large particles of impurities in the sampling liquid and protect the nucleic acid adsorption flow channel 207 from being blocked; the nucleic acid adsorption flow channel Fillers capable of adsorbing nucleic acids are preset in 207, such as a silicon-based membrane. The sampling liquid flow chamber 203, the cleaning liquid flow chamber 204 and the elution liquid flow chamber 205 are all connected to the nucleic acid adsorption flow channel 207, and the nucleic acid adsorption flow channel 207 is connected to the premixing chamber 208. The eluent is pure water, and the eluent enters the premixing chamber 208 through another channel, thereby ensuring the purity of the eluent, preventing it from being contaminated by the sampling liquid and the cleaning liquid, and ensuring the accuracy of the experimental results.

[0054] The reaction liquid flow chamber 206 is connected to the premixing chamber 208, and the premixing chamber 208 is connected to the reaction chamber 209; the premixing negative pressure port 212 is used to draw negative pressure and drive the reaction liquid and the eluent to flow into the premixing chamber 208 for mixing.

[0055] The reaction module 2 is further provided with a cleaning liquid suction port 213 . When the sampling liquid containing cavity 101 is connected to a sampling bottle, when the sampling bottle needs to be cleaned, the cleaning liquid for cleaning the sampling bottle is pumped out from the cleaning liquid suction port 213 to the outside.

[0056] The reagent storage module 1 is provided with a first waste liquid chamber 119, and there are three first waste liquid chambers 119. In order to increase the strength between the waste liquid chambers, partitions are provided between the three first waste liquid chambers 119; in order to simplify the structure, the bottoms of the three first waste liquid chambers 119 are interconnected, and when one of the first waste liquid chambers 119 is filled with waste liquid, it will flow to the other two first waste liquid chambers 119. In order to simplify the structure, a waste liquid connection port 122 is provided behind the waste liquid chamber in the middle of the three first waste liquid chambers 119 (the bottoms of the three waste liquid chambers are interconnected, so only one waste liquid port 211 is required, and the structure is more compact and ingenious than providing three waste liquid ports). Driven by negative pressure, the waste liquid is connected to the waste liquid connection port 122 through an external pipe and finally enters the first waste liquid chamber 119. After the sampling liquid passes through the silicon-based membrane, the nucleic acid is adsorbed on the silicon-based membrane, and the excess sampling liquid is waste liquid; after the cleaning liquid cleans away the impurities on the silicon-based membrane, the nucleic acid is still adsorbed on the silicon-based membrane, the cleaning liquid and impurities are waste liquid, and these waste liquids all enter the first waste liquid chamber 119.

[0057] The reaction module 2 is further provided with a second waste liquid chamber 215 , and the reaction chamber 209 is communicated with the second waste liquid chamber 215 .

[0058] The reaction module 2 is provided with an injection negative pressure port 214, which is connected to the reaction chamber 209; the reaction chamber 209 can be pre-pumped into a negative pressure state through the negative pressure port or by real-time pumping of the injection negative pressure port 214 to drive the mixed liquid in the premixing chamber 208 to flow into the reaction chamber 209, and the excess waste liquid flows into the second waste liquid chamber 215.

[0059] The pre-reaction sampling liquid accommodating cavity, the cleaning liquid accommodating cavity, the eluting liquid accommodating cavity, the first reaction liquid accommodating cavity, each waste liquid cavity, the reaction liquid vent, and the third and second spike portion accommodating channel ports are all sealed with sealing films, and the first spike portion 106 is sealed with an elastic rubber cap to isolate the chamber from the outside world and reduce contamination.

[0060] In the present invention, the cleaning liquid accommodating chamber 102 and the eluting liquid accommodating chamber 103 are vented to the atmosphere through through holes and connecting channels; the cleaning liquid accommodating chamber 102 and the eluting liquid accommodating chamber 103 are pre-filled with cleaning liquid and eluting liquid, and the bottom, surface and top surface thereof are sealed with sealing films. Such a design can not only keep the solution in the accommodating chamber for a long time and stably, but also release the solution in the accommodating chamber in the corresponding experimental steps according to the experimental sequence. In each step, only one corresponding accommodating chamber is vented to the atmosphere and the solution in the accommodating chamber is released. After the solution is released, the control The corresponding containing chamber is isolated from the atmosphere, otherwise the negative pressure cannot drive the liquid to flow when releasing the liquid in the next step, because the flow resistance of the gas is smaller than the flow resistance of the liquid, and the negative pressure will preferentially draw the atmosphere. (The sampling liquid containing chamber and the second reaction liquid containing chamber do not need to be pre-filled with solution. In order to simplify the structure, only a spike portion is required at the bottom and an elastic rubber cap is used to isolate the sampling liquid containing chamber and the second reaction liquid containing chamber from the outside world. However, the cleaning liquid containing chamber 102 and the elution liquid containing chamber 103 pre-filled with solution cannot be isolated from the outside world in this way, and leakage will occur).

[0061] In an embodiment of the present invention, the sampling liquid is used to lyse the pathogenic microorganisms in the sample, and the nucleic acid is released into the sampling liquid. The sampling liquid with the nucleic acid flows through the silicon-based membrane, and the nucleic acid is adsorbed on the silicon-based membrane. The excess sampling liquid waste flows into the waste liquid chamber; the cleaning liquid passes through the membrane to clean the impurities on the silicon-based membrane to prevent the impurities from inhibiting the reaction between the reaction liquid and the nucleic acid; after the nucleic acid is evenly mixed in the premixing chamber and the reaction liquid, it is pumped into the reaction chamber by negative pressure. The reaction chamber is embedded with primers. The nucleic acid, the reaction liquid, and the primer react with color under heating conditions. By observing the color change in the reaction chamber, the positive and negative properties are judged to achieve the detection of pathogenic microorganisms;

[0062] In a specific embodiment, the workflow of the split microfluidic chip is as follows:

[0063] Initial state: the reagent storage module 1 is connected to the connection module 3 through the first buckle 301, and the reaction module 2 is connected to the first limiting groove 201 through the second buckle 302.

[0064] 1. Puncture the sealing film at the top of the sampling liquid containing chamber 101 to connect the sampling liquid containing chamber 101 to the atmosphere, add the sampling liquid into the sampling liquid containing chamber 101, and the sampling liquid is used to lyse the sample, so that the nucleic acid in the sample is released in the lysate.

[0065] External force pushes the reagent storage module 1 to slide in the connecting module 3 until the reaction module 2 is engaged with the second limiting groove 202 through the second buckle 302. At this time, the reagent storage module 1 and the reaction module 2 are abutted, and the first spike portion 106 at the bottom of the sampling liquid accommodating chamber 101 pierces the sealing film at the first interface, and the first spike portion 106 at the bottom of the reaction liquid storage component 104 pierces the sealing film at the fourth interface. The sampling liquid accommodating chamber 101 is connected with the sampling liquid flow chamber 203, and the reaction liquid storage component 104 is connected with the reaction liquid flow chamber 206 (the first spike portion 106 is a hollow structure). Negative pressure is drawn to the waste liquid port 211 (i.e., the negative pressure port), and the sample liquid flows through the sample liquid flow chamber 203, the nucleic acid adsorption flow channel 207, and the first waste liquid chamber 119 in sequence. The nucleic acid in the sample liquid is adsorbed on the nucleic acid adsorbent, and the remaining sample liquid is drawn into the first waste liquid chamber 119 by negative pressure. After the sample liquid is drawn, the float 105 descends to the bottom of the sample liquid accommodating chamber 101 to block the first spike portion 106, and the first spike portion 106 is not connected to the outside.

[0066] 2. The external force drives the third spike portion 110 to slide downward, and the third spike portion 110 drives the second spike portion 108 in the cleaning liquid accommodating chamber 102 to slide downward, and the second spike portion 108 pierces the sealing film at the second docking port 217 until the first through hole 112 slides to the first connecting channel 115. At this time, the third through hole 114 of the third spike portion 110 just slides to the third connecting channel 117, and the cleaning liquid accommodating chamber 102 is connected to the atmosphere through the third through hole 114 and the hollow structure of the third spike portion 110, and the waste liquid port 211 ( The cleaning liquid flows into the hollow structure from the first through hole 112 and then enters the cleaning liquid flow chamber 204, the nucleic acid adsorption flow channel 207, and the first waste liquid chamber 119. The cleaning liquid washes away the impurities on the nucleic acid adsorbent, and the nucleic acid is retained on the nucleic acid adsorbent. After the cleaning liquid is pumped out, the external force continues to drive the fifth spike portion 110 to slide downward until the first through hole 112 slides downward away from the first connecting channel 115, driving the third spike portion 110 to leave the second spike portion accommodating channel. At this time, the cleaning liquid accommodating chamber 102 is not connected to the outside atmosphere.

[0067] 3. External force drives the third spike portion 110 to slide downward, driving the second spike portion 108 in the eluent accommodating chamber 103 to slide downward, and the second spike portion 108 pierces the sealing film at the third interface until the second through hole 113 slides to the second connecting channel 116. At this time, the third through hole 114 of the third spike portion 110 just slides to the fourth connecting channel 118, and the eluent accommodating chamber 103 is connected to the atmosphere through the third through hole 114 and the hollow structure of the third spike portion 110, and the premixed negative pressure port 212 is pumped. Negative pressure, the eluent (i.e., water) flows into the hollow structure from the second through hole 113 and then enters the eluent flow chamber 205, the nucleic acid adsorption channel 207, and the premixing chamber 208, and finally the eluent elutes the nucleic acid adsorbed on the nucleic acid into the premixing chamber 208; after the eluent is exhausted, the external force continues to drive the third spike portion 110 to slide downward until the second through hole 114 slides downward away from the second connecting channel 116, driving the third spike portion 110 to leave the second spike portion accommodating channel, and at this time the eluent accommodating chamber 103 is not connected to the outside atmosphere;

[0068] 4. The reaction liquid storage component 104 on the heating chip is heated to melt the paraffin in the reaction liquid storage component 104. At this time, the first reaction liquid accommodating chamber 1041 is connected to the second reaction liquid accommodating chamber 1043. The air vent 210 is punctured to connect the first reaction liquid accommodating chamber 1041 to the atmosphere. The negative pressure is pumped to the premix negative pressure port 212. The reaction liquid in the first reaction liquid accommodating chamber 1041 flows into the second reaction liquid accommodating chamber 1043 through the S flow channel and is connected to the second reaction liquid accommodating chamber 1043. The reaction liquid in 1043 is mixed, and when the reaction liquid mixed with paraffin drops on the side plate 123, the paraffin solidifies on the side plate 123 when it is cold, and the paraffin is separated from the reaction liquid, thereby preventing the paraffin from solidifying in the premixing chamber 208 and preventing the premixing chamber 208 and the reaction chamber 209 from being unable to communicate due to the paraffin solidifying at the bottom of the premixing chamber 208. The reaction liquid finally flows to the premixing chamber 208, and the mixed reaction liquid in the second reaction liquid containing chamber 1043 finally flows into the premixing chamber 208 under the action of the driving force;

[0069] 5. Negative pressure is applied to the negative pressure port 214. The solution in the premixing chamber 208 flows into each reaction chamber 209 under the action of the negative pressure driving force. The excess liquid enters the second waste liquid chamber 215. The reaction chamber 209 is heated. The nucleic acid and the reaction solution react with color under the heating condition. The positive and negative nature of the sample is determined by the color change.

Claims

1. A split microfluidic chip, characterized in that: It includes a reagent storage module, a reaction module and a connection module; The reagent storage module is connected to the reaction module through the connection module. The connection module is provided with a slide groove, and the reaction module is slidably arranged in the slide groove. When working, the reaction module is pushed to achieve mutual contact between the reaction module and the reagent storage module; wherein: The reagent storage module is provided with a plurality of solution containing chambers, the top openings of the solution containing chambers are isolated from the outside by a sealing film, the bottom openings of some of the solution containing chambers are connected to the first spike portion, the first spike portion is open at both ends and is hollow, the bottom of another part of the solution containing chambers is sealed with a sealing film, a connecting channel is provided near the bottom, the top of the connecting channel is a spike portion containing channel, a second spike portion is slidably provided in the spike portion containing channel, the second spike portion is bounded by a through hole, the upper part is solid, the lower part is hollow and a through hole is provided on the side wall, when an external force is applied, the second spike portion slides downward in the spike portion containing channel so that the through hole on the second spike portion and the connecting channel provided near the bottom of the solution containing chamber are connected to each other; The reaction module is provided with a plurality of solution flow chambers, and a sealing film is provided on the top opening of the solution flow chamber; the number and position of the solution flow chambers correspond to the solution containing chambers in the reagent storage module; when working, the first spike portion and the second spike portion pierce the sealing film at the top of the solution flow chamber, and under the action of negative pressure, the solution in the solution containing chamber is driven to flow to the corresponding solution flow chamber, so as to realize the directional flow of the solution; wherein: On the reagent storage module, the third spike portion is used to apply an external force to the second spike portion arranged in the spike portion accommodating channel so that the second spike portion slides downward, the third spike portion has an open top, a hollow structure at the top and a through hole on the side wall, and a closed sharp portion at the bottom; A connecting channel is arranged on the upper part of the solution containing chamber with the spiked portion accommodating channel arranged inside, and the connecting channel, the spiked portion accommodating channel and the solution containing chamber are all connected, so that during operation, the solution containing chamber and the atmosphere are connected through the connection between the through hole on the third spiked portion and the connecting channel.

2. The split microfluidic chip according to claim 1, characterized in that: A first buckle and a second buckle are provided on the connecting module, a first limiting groove and a second limiting groove are provided on the side wall of the reaction module, a buckle connection is achieved between the reagent storage module and the connecting module via the first buckle, and a fixed connection is achieved between the reaction module and the connecting module in an initial state and a working state respectively via the cooperation between the second buckle and the first limiting groove and the second limiting groove.

3. The split microfluidic chip according to claim 1, characterized in that: The reaction module is also provided with a premixing chamber and a reaction chamber. The solution flow chamber is interconnected with the premixing chamber through a flow channel, and the premixing chamber is connected with the reaction chamber through the flow channel.

4. The split microfluidic chip according to claim 3, characterized in that: The solution containing chamber is used to store sampling liquid, cleaning liquid, eluent and reaction liquid respectively; a float is arranged in the solution containing chamber for storing the sampling liquid, and a filter membrane is arranged in the solution flow chamber connected to the solution containing chamber for storing the sampling liquid to filter large particles of impurities in the sampling liquid; The flow channel connecting the solution flow chamber and the premixing chamber is a nucleic acid adsorption flow channel, in which a filler for adsorbing nucleic acid is arranged; there are several reaction chambers connected in series, and each reaction chamber has the same volume.

5. The split microfluidic chip according to claim 4, characterized in that: The solution containing chamber for storing the reaction liquid includes a plurality of reaction liquid containing chambers interconnected by S-bends, a paraffin block is arranged inside the S-bends for isolating and separating different reaction liquids, and a side plate is arranged inside the reaction liquid containing chamber for receiving the cooled paraffin block.

6. The split microfluidic chip according to claim 5, characterized in that: The reagent storage module and the reaction module are respectively provided with a first waste liquid chamber and a second waste liquid chamber. The first waste liquid chamber is used to collect excess sampling liquid and cleaning liquid through an external pipe, and the second waste liquid chamber is used to collect excess waste liquid flowing from the premixing chamber to the reaction chamber. Negative pressure ports are respectively provided on the reagent storage module and the reaction module, and the negative pressure ports are respectively connected to the first waste liquid chamber, the premixing chamber, and the second waste liquid chamber through external pipes and flow channels to realize directional flow of the solution in the chip and achieve sample sampling, cleaning, nucleic acid adsorption, elution and reaction mixing reactions.

7. A method for operating the split microfluidic chip according to claim 6, characterized in that: The method includes the following two solution directional flow steps: (1) External force pushes the reaction module to abut the reaction module and the reagent storage module, and the first spike portion connected to the bottom of the solution containing chamber pierces the sealing film at the top of the corresponding solution flow chamber, thereby realizing the mutual communication between the solution containing chamber and the solution flow chamber. At the same time, through the communication between the top of the solution containing chamber and the atmosphere, under the action of negative pressure, the solution in the solution containing chamber is driven to flow to the corresponding solution flow chamber; (2) The external force drives the third spike portion to slide downward, and drives the second spike portion in the spike portion accommodating channel to slide downward to pierce the sealing film at the bottom of the corresponding solution accommodating chamber and the sealing film at the top of the flow chamber. The solution accommodating chamber and the solution flow chamber are connected to each other through the through hole on the second spike portion and the connecting channel arranged near the bottom of the accommodating chamber. At the same time, the solution accommodating chamber is connected to the atmosphere through the through hole of the third spike portion at the upper part of the solution accommodating chamber and the connecting channel at the upper part of the solution accommodating chamber. Under the action of negative pressure, the solution in the solution accommodating chamber is driven to flow to the corresponding solution flow chamber.

8. The working method according to claim 7, characterized in that: The following steps are involved: Step 1: external force pushes the reaction module to abut the reaction module and the reagent storage module, and at this time, the solution containing chamber for storing the sampling liquid, the solution containing chamber for storing the reaction liquid and the corresponding solution flow chamber are respectively connected; at the same time, the sealing film on the top of the containing chamber for storing the sampling liquid is connected to the atmosphere, and under the action of negative pressure, the sampling liquid flows through the corresponding solution flow chamber, the nucleic acid adsorption channel and the first waste liquid chamber in sequence; after the sampling liquid is drawn out, the float drops to the bottom of the containing chamber to block the spike part, and the first spike part is not connected to the outside atmosphere; Step 2: external force drives the third spike portion to slide downward, driving the second spike portion in the solution containing chamber storing the cleaning liquid to slide downward to pierce the sealing film at the top of the corresponding solution flow chamber, and the through hole on the second spike portion is connected to the connecting channel arranged near the bottom of the solution containing chamber. At this time, the solution containing chamber storing the cleaning liquid is connected to the atmosphere through the through hole on the third spike portion above. Under the action of negative pressure, the cleaning liquid enters the corresponding solution flow chamber, the nucleic acid adsorption channel, and the first waste liquid chamber in turn; after the cleaning liquid is pumped out, the external force continues to drive the second spike portion to slide downward so that the solution containing chamber is not connected to the outside atmosphere; Step 3: external force drives the third spike portion to slide downward, driving the second spike portion in the solution containing chamber storing the eluent to slide downward to pierce the sealing film at the top of the corresponding solution flow chamber, and the through hole on the second spike portion is connected to the connecting channel arranged near the bottom of the solution containing chamber. At this time, the solution containing chamber storing the eluent is connected to the atmosphere through the through hole of the third spike portion above. Under the action of negative pressure, the cleaning liquid enters the corresponding flow chamber, the nucleic acid adsorption flow channel, and the premixing chamber; after the cleaning liquid is pumped out, the external force continues to drive the second spike portion to slide downward so that the containing chamber is not connected to the outside atmosphere; Step 4: Connect the solution containing chamber storing the reaction solution to the atmosphere, and under the action of negative pressure, the reaction solution flows into the premixing chamber; Step 5: The solution in the premixing chamber flows into the reaction chamber under the action of negative pressure, and the excess liquid enters the second waste liquid chamber.

Images