WO2020216058A1

WO2020216058A1 - Multi-index test microfluidic kit and application method therefor

Info

Publication number: WO2020216058A1
Application number: PCT/CN2020/083549
Authority: WO
Inventors: 徐友春; 李楠
Original assignee: 清华大学
Priority date: 2019-04-23
Filing date: 2020-04-07
Publication date: 2020-10-29
Also published as: CN110075935B; CN110075935A

Abstract

Disclosed is a multi-index test microfluidic kit and an application method therefor. The kit comprises: a piston pump (100), liquid storage tubes (200), a liquid storage tube cover (300), a kit base (400), a cover plate (500), a sealing gasket (600), a gating valve (700) and a base plate (800), wherein the piston pump (100) drives kit fluid in a push and pull manner, the gating valve (700) rotates to open and close a flow channel connection, the liquid storage tubes (200) store reagent samples, the liquid storage tube cover (300) seals the liquid storage tubes (200), the kit base (400) supports and is connected to all parts, the cover plate (500) seals a flow path pipeline and a cavity in the kit base (400), the sealing gasket (600) is mounted between the kit base (400) and the gating valve (700) for sealing, and the base plate (800) assembles the gating valve (700) to the kit base (400) by means of a fixing component.

Description

Multi-index detection microfluidic cartridge and application method

Cross references to related applications

This application claims the priority of the Chinese patent application number "201910329723.9" submitted by Tsinghua University on April 23, 2019, with the title of "Multi-indicator Detection Microfluidic Cartridge and Application Method".

Technical field

The invention relates to the technical field of in vitro diagnosis, in particular to a multi-index detection microfluidic cartridge and an application method.

Background technique

Fully integrated automated analysis is the development trend in the field of in vitro diagnostics. For molecular diagnosis in in vitro diagnostics, integrated automated nucleic acid detection technology is the development trend of molecular diagnostic technology. Its important significance is not only to solve the problem of manpower and laboratory space occupation, but also to avoid the harm of samples to operators and step-by-step operations Bringing problems such as potential product contamination can effectively improve the reliability of clinical diagnosis and reduce costs. For immunological and biochemical diagnosis in in vitro diagnostics, fully automatic multi-indicator joint detection is the development direction. It can not only save manpower and material resources, but also provide doctors with abundant clinical test information. The optimized combination of multiple indicators can improve the detection rate of diseases , Reduce missed diagnosis and misdiagnosis.

The three basic types of in vitro diagnostic reactions require a wide variety of reagents, complicated steps, and complex liquid manipulation and detection devices. Therefore, instruments and equipment used for in-vitro diagnostic testing often require more liquid control components, making the equipment larger in size and difficult to meet the needs of real-time testing in areas with backward medical infrastructure and on-site. In addition, the sample processing process often cannot be flexibly changed according to actual needs, and the types and volume ranges of processed samples are small.

Summary of the invention

The present invention aims to solve one of the technical problems in the related art at least to a certain extent.

To this end, an object of the present invention is to provide a multi-index detection microfluidic cartridge, which can fully integrate and automatically process samples, and better meet the needs of in vitro diagnostic testing.

Another object of the present invention is to provide an application method of the multi-index detection microfluidic cartridge.

In order to achieve the above objective, an embodiment of the present invention proposes a multi-index detection microfluidic cartridge, which includes: a piston pump, the piston pump and the gate valve are sealed connected by an interface for driving the cartridge fluid; The liquid tube is used to store samples, reagents and waste liquid, wherein the bottom end of the liquid storage tube is provided with a through hole, which is connected to the base of the cartridge through an upper interface; the liquid storage tube cover, the liquid storage tube cover and the The liquid storage tube is hermetically connected, wherein the liquid storage tube cover is provided with a vent hole smaller than the diameter of the inner wall of the liquid storage tube; the cartridge base is provided with the liquid storage tube connection interface and multiple indicators The detection reaction chamber and its non-interference injection pipe and exhaust pipe, the through hole communicating with the gate valve, the gate valve placement groove, the bottom plate fixing interface, the multi-index detection reaction chamber is fixed There are freeze-dried detection reagents, the through holes are circumferentially divided; a cover plate, the cover plate is located above the cartridge base to adhesively seal the multi-index reaction chamber of the cartridge base and its sampling pipeline and Exhaust pipeline; a gasket, the gasket is located on the top surface of the gate valve placement groove of the cartridge base; a gate valve, the gate valve is located below the gasket, and the gate valve is located at the cartridge base In the placement groove, the gate valve is provided with a piston pump interface, a fluid pipeline, an outlet, a vent hole, and a clamping groove connected to an external instrument, and the piston pump interface and the fluid pipeline communicate with the outlet The vent hole penetrates the gate valve and is separated from the fluid pipe and the outlet by a certain distance. By controlling the slot where the gate valve is connected to an external instrument, the gate valve outlet is connected to the The liquid storage tube and the multi-index detection reaction chamber sample injection pipeline are gated; a bottom plate, the bottom plate is under the gate valve, and the cartridge base is assembled with the cartridge via a fixing member.

The multi-index detection microfluidic cartridge of the embodiment of the present invention can perform multi-index automatic detection and analysis of nucleic acid, protein and biochemistry on biological samples. The operation process is simple, the test results are rich, and it has high clinical application value and is universal The performance is greatly enhanced, and it can meet the testing needs of different types of clinical samples more widely. It has the advantages of full integration, higher testing stability and efficiency, and lower cost. Users use the same hardware system for different types of testing. It is helpful to improve user acceptance and promote the popularization of in vitro diagnosis.

In addition, the multi-index detection microfluidic cartridge according to the foregoing embodiment of the present invention may also have the following additional technical features:

Further, in an embodiment of the present invention, the piston pump includes a push-pull piston rod and a piston barrel to control the up and down movement of the piston rod according to a control signal of an external instrument.

Further, in an embodiment of the present invention, the number of the liquid storage tube is at least two.

Further, in an embodiment of the present invention, a hydrophobic gas-permeable membrane is provided at the position of the vent hole of the liquid storage tube cover.

Further, in an embodiment of the present invention, the number of the multi-index detection reaction chamber of the cartridge base is at least one.

Further, in an embodiment of the present invention, the bottom of the multi-index detection reaction chamber of the cartridge base is provided with a notch for fixing the flat base.

Further, in an embodiment of the present invention, the cover plate is made of a transparent material to obtain a biological detection response.

Further, in an embodiment of the present invention, the sealing gasket has the same shape and size as the gate valve placement groove of the cartridge base, and the through hole of the cartridge base is provided with Through hole.

Further, in an embodiment of the present invention, the cartridge base is made of transparent plastic, and the sealing gasket is made of elastic polymer material.

In order to achieve the above objective, another embodiment of the present invention proposes an application method for a multi-index detection microfluidic cartridge, which adopts the multi-index detection microfluidic cartridge as described in the above embodiment, wherein the method includes: Step S1, the reagents required for the reaction are stored in the liquid storage tube of the cartridge in advance, and after the user adds the sample to the liquid storage tube, the cartridge is placed in the supporting control instrument; Step S2, external control is used The instrument is controlled by the cooperation of the piston pump and the gate valve to transfer, mix, and distribute the multi-step fluid, and complete the required biological detection reaction according to the target temperature control and magnetic bead control of the detection reaction Step S3, controlling the external control instrument to read the reaction result of the multi-index reaction chamber of the cartridge.

The application method of the multi-index detection microfluidic cartridge of the embodiment of the present invention can perform multi-index automatic detection and analysis of nucleic acid, protein, and biochemistry on biological samples. The operation process is simple, the test results are rich, and it has high clinical application value. , And the versatility is greatly enhanced, and it can meet the testing needs of different types of clinical samples more widely. It has the advantages of full integration, higher testing stability and efficiency, and lower cost. Users use the same set of testing for different types of testing. The hardware system is conducive to improving user acceptance and promoting the popularization of in vitro diagnosis.

The additional aspects and advantages of the present invention will be partly given in the following description, and partly will become obvious from the following description, or be understood through the practice of the present invention.

Description of the drawings

The above and/or additional aspects and advantages of the present invention will become obvious and easy to understand from the following description of the embodiments in conjunction with the accompanying drawings, in which:

Fig. 1 is a schematic structural diagram of a multi-index detection microfluidic cartridge according to an embodiment of the present invention;

2 is a schematic diagram of the structure of a multi-index detection microfluidic cartridge according to the first embodiment of the present invention;

3a and 3b are schematic diagrams of the structure of the cartridge base according to the first embodiment of the present invention;

Figure 4 is a schematic structural diagram of a gate valve according to a first embodiment of the present invention;

5a and 5b are cross-sectional views of the gate valve and the gated liquid storage tank according to the first embodiment of the present invention;

6a to 6o are schematic diagrams of the gated flow path of the gate valve at different stages of operation of the cartridge according to the first embodiment of the present invention;

Figure 7 is a schematic diagram of the electromagnet operating the magnetic beads of the cartridge according to the first embodiment of the present invention;

8a and 8b are sectional views of a liquid storage tube equipped with a silicone mold according to a second embodiment of the present invention;

Fig. 9 is a schematic structural diagram of a multi-index detection microfluidic cartridge according to a third embodiment of the present invention;

Figure 10 is a schematic diagram of a gate valve and a gate flow path according to a third embodiment of the present invention;

11 is a schematic diagram of the structure of a multi-index detection microfluidic cartridge according to a fourth embodiment of the present invention;

Fig. 12 is a flowchart of an application method for multi-index detection of a microfluidic cartridge according to an embodiment of the present invention.

Detailed ways

The embodiments of the present invention are described in detail below. Examples of the embodiments are shown in the accompanying drawings, in which the same or similar reference numerals indicate the same or similar elements or elements with the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary, and are intended to explain the present invention, but should not be construed as limiting the present invention.

The following describes the multi-index detection microfluidic cartridge and application method according to the embodiments of the present invention with reference to the accompanying drawings. First, the multi-index detection microfluidic cartridge proposed according to the embodiments of the present invention will be described with reference to the accompanying drawings.

Fig. 1 is a schematic structural diagram of a multi-index detection microfluidic cartridge according to an embodiment of the present invention.

As shown in Figure 1, the multi-index detection microfluidic cartridge includes: a piston pump 100, a liquid storage tube 200, a liquid storage tube cover 300, a cartridge base 400, a cover plate 500, and a gasket 600 (not shown in Figure 1 Specifically labeled), gate valve 700 (not specifically labeled in FIG. 1), and bottom plate 800.

Wherein, the piston pump 100 and the gate valve 700 are connected in a sealed manner through an interface, and are used to drive the cartridge fluid. The liquid storage tube 200 is used to store samples, reagents, and waste liquid. The bottom end of the liquid storage tube 200 is provided with a through hole and is connected to the cartridge base 400 in a sealed manner through an upper interface. The liquid storage tube cover 300 is in sealed connection with the liquid storage tube 200, wherein the liquid storage tube cover 300 is provided with a vent hole smaller than the diameter of the inner wall of the liquid storage tube 200. The cartridge base 400 is provided with a liquid storage pipe 200 connection interface, a multi-index detection reaction chamber and its non-interference injection and exhaust pipes, a through hole communicating with the gate valve 700, a slot for the gate valve 700, and a fixed bottom plate Interface, multi-index detection reaction chamber is fixed with freeze-dried detection reagents, and the through holes are circular sections. The cover plate 500 is located above the cassette base 400 to bond and seal the multi-index reaction chamber of the cassette base 400 and its sample inlet pipe and exhaust pipe. The sealing gasket 600 is located on the top surface of the slot of the cartridge base 400 and the gate valve 700. The gate valve 700 is located under the sealing gasket 600, in the cartridge base 400, the gate valve 700 is placed in the groove, the gate valve 700 is provided with piston pump interfaces, fluid pipes, outlets, vents and cards connected to external instruments The port of the piston pump 100, the fluid pipeline and the outlet are connected, the vent hole penetrates the gate valve 700 and is separated from the fluid pipeline and the outlet by a certain distance, and the gate valve 700 is connected to the slot by controlling the gate valve 700 and the external instrument. The outlet and the liquid storage pipe 200 and the multi-index detection reaction chamber injection pipe are gated. The bottom plate 800 is under the gate valve 700, and the cartridge base 400 is assembled with the cartridge via a fixing member. The cartridge of the embodiment of the present invention can perform multi-index automatic detection and analysis of nucleic acid, protein, and biochemistry on biological samples, has a simple operation process, abundant test results, and has high clinical application value.

It should be noted that the multi-index detection microfluidic cartridge can be used in biological testing or medical testing. The application of the cartridge in biological testing or medical testing will be described in detail below through examples.

In the first embodiment of the present invention, the application of the cartridge of the embodiment of the present invention in the field of fully integrated nucleic acid is to extract nucleic acid from a sample by a magnetic bead-based method and complete multi-index nucleic acid fragment amplification detection.

Wherein, as shown in Figures 2 to 5, the piston pump 100 includes a push-pull piston rod 101 and a piston barrel 102. The up and down movement of the piston rod 101 is controlled by an external instrument, and is connected to the gate valve 700 through an interface 103 in a sealed manner to drive the cartridge fluid . The liquid storage tubes 200a to 200f are used to store samples, reagents, and waste liquids. The bottom end is provided with a through hole 201, and is connected to the cartridge base 400 through the upper interfaces 401a to 401f. The liquid storage tube cover 300 is in sealed connection with the liquid storage tubes 200a to 200f, and is provided with a vent hole 301 smaller than the diameter of the inner wall of the liquid storage tube 300, and a hydrophobic gas-permeable membrane 302 is provided at the vent hole 301. The cartridge base 400 is provided with liquid storage pipe connection interfaces 401a to 401f, a multi-index detection reaction chamber 402 and its non-interference injection pipe 403 and exhaust pipe 404, through holes 405a to 405c communicating with the gate valve 700, and selection The through valve placement groove 406, the bottom plate fixing interfaces 407a to 407c, and the through holes 405a to 405c are circumferentially divided. The cover plate 500 is located above the cassette base 400 and seals the multi-index reaction chamber 402 of the cassette base and its sampling pipe 403 and exhaust pipe 404. The gasket 600 is located on the top surface of the gate valve placement groove 406 of the cartridge base. The gate valve 700 is located under the sealing gasket 600. In the gate valve placement groove 406 of the cartridge base, there are provided with a piston pump interface 701, a fluid pipe 702, an outlet 703, a vent 704 and a card slot 705 connected with external instruments. , The piston pump interface 701, the fluid pipe 702 and the outlet 703 are in communication, the vent 704 penetrates the gate valve 700 and is separated from the fluid pipe 702 and the outlet 703 by a certain distance, and controls the gate valve 700 to connect with the card slot 705 of the external instrument. The gate valve outlet 703 is gated with the liquid storage pipe 200 and the multi-index detection reaction chamber injection pipe 403. The bottom plate 800 is below the gate valve 700, and the cartridge base 400 is assembled with the cartridge via fixing members 801a to 801d.

Wherein, in an embodiment of the present invention, the cartridge base 400 may be a transparent plastic, and the sealing gasket 600 may be an elastic polymer material. Of course, other types of materials can also be selected for the cartridge base 400 and the sealing gasket, which is only used as an example here and is not specifically limited.

Further, in an embodiment of the present invention, the number of liquid storage tubes 200 is at least two, and the number of the cartridge base multi-index detection reaction chamber 402 is at least one. Those skilled in the art can set the liquid storage tube according to actual conditions. The specific number of the tube 200 and the multi-index detection reaction chamber 402 of the cartridge base is not specifically limited here.

It should be noted that the reagents for nucleic acid extraction and amplification are stored in the storage tubes 200a to 200f in advance, and primers and other substrates are pre-fixed in the multi-index reaction chamber 402.

Further, in an embodiment of the present invention, the cover plate 500 may be a transparent material to obtain a biological detection response.

It is understandable that the cover plate 500 may be a transparent material, compatible with the biological detection reaction, so as to facilitate the detection of the reaction result. Of course, other types of materials can also be selected for the cover plate, which is only used as an example here and is not specifically limited.

Further, in an embodiment of the present invention, the sealing gasket 600 and the cartridge base gate valve placement groove 406 have the same size, and a through hole 601 of the cartridge base is provided at the through hole position of the cartridge base 400.

Further, the process of nucleic acid detection using the cassette of this embodiment is as follows:

1) Add the sample to be tested (such as blood, sputum, saliva, urine, etc.) into the original sample liquid storage tube 200a, and the external instrument controls the gate valve 700 to gate the liquid storage tube 200a (shown in Figure 6a), At this time, the gate valve outlet 703 is aligned with the through hole 405c of the cartridge base (shown in Figures 5a-b), the gate valve through hole 704 is in a closed state, and the piston pump 100 draws a certain amount of sample from the liquid storage tube 200a;

2) After the sample is drawn, the gate valve 700 rotates clockwise to gate the second liquid storage tube 200b containing the lysis solution (shown in Figure 6b). The piston pump 100 quickly pulls several times to achieve full mixing of the sample and the lysis solution;

3) After the sample is lysed, the gate valve 700 rotates clockwise to gate the third liquid storage tube 200c containing magnetic beads (shown in Figure 6c). The piston pump 100 quickly pulls several times to achieve the full mixing of the sample lysis solution and the magnetic bead mixture. After the sample nucleic acid is adsorbed by the magnetic beads, the electromagnet 900 under the control card box is opened (shown in Figure 7), and the piston is slowly pushed The pump 100 pushes the waste liquid into the third storage tube 200c, and the magnetic beads are concentrated directly below the gate valve 700;

4) After the enrichment of the magnetic beads is completed, the gate valve 700 rotates clockwise to gate the fourth liquid storage tube 200d (shown in Figure 6d) containing the cleaning solution, and the electromagnet 900 under the control card box is closed, and it is quickly drawn Pull several times to realize the resuspension of the magnetic beads and fully mix with the cleaning liquid. After the magnetic beads are cleaned by the cleaning liquid, the electromagnet 900 under the control card box is turned on, and the piston pump 100 is slowly pushed to push the waste liquid into the fourth storage. In the tube 200d, the magnetic beads are concentrated directly under the gate valve 700;

5) The gate valve 700 rotates clockwise to gate the fifth liquid storage tube 200e (shown in Figure 6e) containing the cleaning solution, and repeat step (4) to clean the magnetic beads a second time;

6) After completing the cleaning of the magnetic beads, the gate valve 700 rotates clockwise to gate the sixth liquid storage tube 200f (shown in Figure 6f) containing the eluent, and the electromagnet 900 under the control card box is closed for rapid extraction. Pull the piston pump 100 several times to resuspend the magnetic beads and fully mix with the eluent;

6) After the nucleic acid elution is completed, the electromagnet 900 under the control cassette is turned on, and the nucleic acid solution is sequentially injected into the multi-index reaction chamber 402 (shown in Figure 6g-n). At this time, the gate valve outlet 703 and the cassette base The through holes 405a are aligned, and the gate valve vent 704 is aligned with the through holes 405b of the cartridge base;

7) After the sample addition is completed, rotate the cartridge gating valve 700 so that the gating valve outlet 703 and the vent 704 are sealed, and the multi-index reaction chamber 402 is physically and completely isolated (shown in Figure 6i), and finally the supporting control is started The heating control of the instrument realizes multi-index amplification. Here, variable temperature PCR amplification can be used, or constant temperature amplification can be used. The corresponding fluorescence detection system scans and detects the fluorescence signal value of the multi-index reaction chamber 402 in real time, and interprets the detection result.

The above is a detailed description of the first embodiment of the present invention. In the second embodiment of the present invention, as shown in Figures 8a-b, the difference between this embodiment and the first embodiment is that this embodiment uses a silicone mold-based In the nucleic acid extraction scheme, the silicone membrane 1000 is fixed at the bottom of the liquid storage tube 200c, and the fluid control process is different.

1) Add the sample to be tested (such as blood, sputum, saliva, urine, etc.) into the liquid storage tube 200a containing the lysis solution, and the external instrument controls the gating valve 700 to gate the liquid storage tube 200a (as shown in Figure 6a). Show), at this time, the gate valve outlet 703 is aligned with the through hole 405c of the cartridge base, the gate valve through hole 704 is in a closed state, and the piston pump 100 is quickly pulled several times to achieve sufficient mixing of the sample and the lysis solution;

2) After the sample is lysed, the gate valve 700 rotates clockwise to gate the second storage tube 200b containing the binding buffer (shown in Figure 6b). The piston pump 100 pulls several times and incubates to achieve full mixing of the sample lysis solution and the binding buffer;

3) After that, the gate valve 700 rotates clockwise to gate the third liquid storage tube 200c (shown in Figure 6c) fixed with the silicone mold 1000, and the piston pump 100 pushes the solution into the liquid storage tube 200c, and then slowly pushes the solution Draw back into the piston pump 100 to realize the enrichment and capture of nucleic acid in the silica gel mold 1000;

4) After the nucleic acid enrichment is completed, the gate valve 700 rotates counterclockwise to gate the second liquid storage tube 200b (shown in Figure 6b), and the piston pump 100 discharges the waste liquid into the second liquid storage tube 200b;

5) The gate valve 700 rotates clockwise to gate the fourth liquid storage tube 200d (shown in Figure 6d) containing the cleaning liquid, the piston pump 100 draws the cleaning liquid, and then the gate valve 700 rotates counterclockwise to gate the third The liquid storage tube 200c (shown in Figure 6c) is slowly pulled several times to clean the silicone mold for the first time. The cleaned waste liquid is pumped into the piston pump 100, and the gate valve 700 is turned clockwise to gate the fourth The liquid storage pipe 200d (shown in Figure 6d), the piston pump 100 drains the waste liquid;

6) Rotate the gate valve 700 clockwise to gate the fifth liquid storage tube 200e (shown in Figure 6e) containing the cleaning solution, repeat step (5) to clean the magnetic beads a second time, and drain the waste liquid Into the liquid storage pipe 200e;

7) After completing the two-step nucleic acid cleaning, the gate valve 700 rotates clockwise to gate the sixth liquid storage tube 200f (shown in Figure 6f) containing the eluent, the piston pump 100 draws the eluent, and rotates counterclockwise to select The through valve 700 gates the third liquid storage tube 200c (shown in FIG. 6c) to quickly pull the piston pump 100 to achieve the elution of nucleic acid;

8) The piston pump 100 extracts the nucleic acid eluate and distributes it into the multi-index reaction chamber 402 for multi-index amplification analysis and detection.

In the third embodiment of the present invention, as shown in Figures 9 and 10, the difference between this embodiment and the first embodiment is that: this embodiment uses the cartridge in the field of protein immunoassay, and the multi-index detection of the cartridge base 400 The bottom of the reaction chamber 402 is provided with a notch 408 for fixing the planar substrate, and the protein matrix chip 1100 is fixed, and the fluid control process is different.

Further, the immobilized protein matrix chip 1100 is preferably made of glass. After being modified with agarose or epoxy groups, the sample is coated with a primary antibody solution, and the antibody used for capture is fixed on the surface of the protein matrix chip, and then BSA is performed. Close the chip.

Further, the prepared protein dot matrix chip 1100 is fixedly installed in the notch 408 of the fixed plane base of the cassette base.

Further, the process of using the cassette of this embodiment to perform multi-index protein detection is as follows:

1) Add the sample to be tested (such as serum, etc.) into the first sample liquid storage tube 200a, the external instrument controls the gate valve 700 to gate the liquid storage tube 200a, and the piston pump 100 draws a certain amount from the liquid storage tube 200a After the gate valve 700 rotates clockwise to gate the multi-index reaction chamber 402, the gate valve outlet 703 is aligned with the cartridge base through hole 405a, and the gate valve vent 704 is aligned with the cartridge base through hole 405b Accurately, the piston pump 100 pulls several times to realize that the antigen in the sample is fully combined with the primary antibody coated on the protein matrix chip 1100, and then drain the liquid in the multi-index reaction chamber 402, and rotate the gate valve 700 clockwise to select Pass the liquid storage pipe 200a, and then the piston pump 100 pushes the liquid completely into the liquid storage pipe 200a;

2) The gate valve 700 rotates clockwise to gate the second liquid storage tube 200b containing cleaning liquid, the piston pump 100 will draw a certain amount of cleaning liquid from the liquid storage tube 200b, and then the gate valve 700 rotates clockwise to select Pass the multi-index reaction chamber 402, pull the piston pump 100 several times to fully clean the protein dot matrix chip 1100, then drain the liquid in the multi-index reaction chamber 402, and rotate the gate valve 700 clockwise to gate the liquid storage tube 200b , Then the piston pump 100 completely pushes the liquid into the liquid storage tube 200b;

3) The gate valve 700 rotates clockwise to gate the third liquid storage tube 200c equipped with the primary antibody. The piston pump 100 will draw a certain amount of primary antibody solution from the liquid storage tube 200c, and then the gate valve 700 rotates clockwise to select Pass through the multi-index reaction chamber 402, pull the piston pump 100 several times to realize the full combination of the primary antibody of the protein lattice chip 1100, then drain the liquid in the multi-index reaction chamber 402, and rotate the gate valve 700 clockwise to gate the liquid Pipe 200c, after which the piston pump 100 pushes the liquid completely into the liquid storage pipe 200c;

4) The gate valve 700 rotates clockwise to gate the fourth liquid storage tube 200d with cleaning liquid, and repeat the process (2);

5) The gate valve 700 rotates clockwise to gate the fifth liquid storage tube 200e equipped with the signal marked secondary antibody. The piston pump 100 will draw a certain amount of the secondary antibody solution from the liquid storage tube 200e, and then the gate valve 700 turns Rotate clockwise to gate the multi-index reaction chamber 402. The piston pump 100 pulls several times to achieve full combination of the protein matrix chip 1100 secondary antibody. After that, drain the liquid in the multi-index reaction chamber 402 and turn the gate valve 700 clockwise to select Pass the liquid storage pipe 200e, and then the piston pump 100 pushes the liquid completely into the liquid storage pipe 200c;

6) Rotate the gate valve 700 clockwise to gate the sixth liquid storage tube 200f containing the cleaning liquid, and repeat the process (2);

7) Perform signal detection of protein dot matrix chip 1100 and interpretation and analysis of results.

In addition, in the same way, the technical solution of this embodiment can also be used for detection of nucleic acid hybridization chips.

In the fourth embodiment of the present invention, as shown in FIG. 11, the difference between this embodiment and the first embodiment is that the cartridge is used in the field of biochemical detection in this embodiment, and only a simple sample dilution step is required. The detection of multiple biochemical indicators is directly performed in the multiple indicator detection cavity 402.

Further, the biochemical reaction substrates of the multi-index reaction chamber are different freeze-dried blood biochemical indexes and detection reagents, including alanine aminotransferase, aspartate aminotransferase, glutamyl transpeptidase, alkaline phosphatase, cholinesterase, Alanine aminotransferase, aspartate aminotransferase, gamma-guamyltransferase, total protein, albumin, glycated albumin, total bilirubin, direct bilirubin, ammonia, urea, creatinine, Uric acid, glucose, total cholesterol, triglycerides, high density lipoprotein, low density lipoprotein, very low density lipoprotein, apolipoprotein A1, apolipoprotein B, calcium, chlorine, iron, phosphorus, sodium, potassium, carbon dioxide .

Further, the process of using the cassette of this embodiment to perform multi-index biochemical detection is as follows:

1) Add the sample to be tested (such as serum, plasma, etc.) into the first sample storage tube 200a, the external instrument controls the gate valve 700 to gate the liquid storage tube 200a, and the piston pump 100 draws from the liquid storage tube 200a Quantitative sample, then the gate valve 700 rotates clockwise to gate the liquid storage tube 200c, at this time the piston pump 100 pushes the sample completely into the liquid storage tube 200c;

2) The gate valve 700 rotates counterclockwise to gate the second liquid storage tube 200b containing the diluent, the piston pump 100 will draw a certain amount of cleaning liquid from the liquid storage tube 200b, and then the gate valve 700 rotates clockwise to gate The liquid storage tube 200c is pulled several times by the piston pump 100 to achieve full mixing of the sample and the diluent;

3) The piston pump 100 quantitatively extracts the solution from the liquid storage tube 200c, and then the gate valve 700 rotates clockwise to select, and the solution is sequentially injected into the multi-index reaction chamber 402 through the precise coordination of the piston pump 100 and the gate valve 700, Different sampling pipes 403 and exhaust pipes 404 are isolated from each other, which eliminates the possibility of cross contamination.

4) Perform signal detection of the multi-index reaction chamber 402 and interpretation and analysis of the results.

According to the multi-index detection microfluidic cartridge provided by the embodiment of the present invention, it can perform multi-index automatic detection and analysis of nucleic acid, protein, and biochemistry on biological samples. The operation process is simple, the test results are rich, and it has high clinical application value. And the versatility is greatly enhanced, and it can meet the testing needs of different types of clinical samples more widely. It has the advantages of full integration, higher testing stability and efficiency, and lower cost. Users use the same set of hardware for different types of testing. The system is conducive to improving user acceptance and promoting the popularization of in vitro diagnosis.

Next, the application method of the multi-index detection microfluidic cartridge according to the embodiment of the present invention will be described with reference to the accompanying drawings.

Fig. 12 is a flowchart of an application method of a multi-index detection microfluidic cartridge according to an embodiment of the present invention.

As shown in FIG. 12, the application method of the multi-index detection microfluidic cartridge adopts the multi-index detection microfluidic cartridge of the foregoing embodiment, wherein the method includes:

Step S1, the reagents required for the reaction are stored in the liquid storage tube of the cartridge in advance, and after the user adds the sample to the liquid storage tube, the cartridge is placed in the supporting control instrument.

Step S2: Use an external control instrument to transfer, mix, and distribute the multi-step fluid through the cooperation of the piston pump and the gate valve. According to the target temperature control and magnetic bead control of the detection reaction, the required biological Check the reaction.

Step S3, controlling the external control instrument to read the reaction result of the multi-index reaction chamber of the cassette.

It should be noted that the foregoing explanation of the embodiment of the multi-index detection microfluidic cartridge is also applicable to the application method of the multi-index detection microfluidic cartridge of this embodiment, and will not be repeated here.

According to the application method of the multi-indicator detection microfluidic cartridge provided by the embodiment of the present invention, the multi-indicator automatic detection and analysis of nucleic acid, protein, and biochemistry can be performed on biological samples. The application value and versatility are greatly enhanced. It can meet the testing needs of different types of clinical samples more widely. It has the advantages of full integration, higher testing stability and efficiency, and lower cost. Users use different types of testing. The same hardware system will help improve user acceptance and promote the popularization of in vitro diagnostics.

In addition, the terms "first" and "second" are only used for descriptive purposes, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of indicated technical features. Therefore, the features defined with "first" and "second" may explicitly or implicitly include at least one of the features. In the description of the present invention, "a plurality of" means at least two, such as two, three, etc., unless otherwise specifically defined.

In the description of this specification, descriptions with reference to the terms "one embodiment", "some embodiments", "examples", "specific examples", or "some examples" etc. mean specific features described in conjunction with the embodiment or example , Structure, materials or features are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the described specific features, structures, materials or characteristics can be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art can combine and combine the different embodiments or examples and the characteristics of the different embodiments or examples described in this specification without contradicting each other.

Although the embodiments of the present invention have been shown and described above, it can be understood that the above-mentioned embodiments are exemplary and should not be construed as limiting the present invention. Those of ordinary skill in the art can comment on the foregoing within the scope of the present invention. The embodiment undergoes changes, modifications, substitutions and modifications.

Claims

A multi-index detection microfluidic cartridge, which is characterized in that it comprises:

A piston pump, the piston pump and the gate valve are connected in a sealed manner through an interface and used to drive the cartridge fluid;

The liquid storage tube is used to store samples, reagents and waste liquid, wherein the bottom end of the liquid storage tube is provided with a through hole, and is connected to the base of the cartridge in a sealed manner through an upper interface;

A liquid storage tube cover, the liquid storage tube cover is connected to the liquid storage tube in a sealed manner, wherein the liquid storage tube cover is provided with a vent hole smaller than the diameter of the inner wall of the liquid storage tube;

The cartridge base is provided with the liquid storage pipe connection interface, the multi-indicator detection reaction chamber and its non-interference injection pipe and exhaust pipe, the through hole communicating with the gate valve, the A gate valve placement slot and a bottom plate fixing interface, a freeze-dried detection reagent is fixed in the multi-index detection reaction chamber, and the through hole is a circumferential division;

A cover plate, the cover plate is located above the cartridge base to adhesively seal the multi-index reaction chamber of the cartridge base and its sampling pipe and exhaust pipe;

A gasket, the gasket is located on the top surface of the cartridge base and the gate valve placement groove;

The gate valve, the gate valve is located under the sealing gasket, in the cartridge base, the gate valve placement groove, the gate valve is provided with the piston pump interface, fluid pipeline, outlet, communication An air hole and a card slot connected with an external instrument, the piston pump interface, the fluid pipe communicate with the outlet, and the vent hole penetrates the gate valve and is separated from the fluid pipe and the outlet by a certain distance , By controlling the slot where the gate valve is connected to the external instrument, the gate valve outlet is gated to the liquid storage pipe and the multi-index detection reaction chamber injection pipe; and

A bottom plate, the bottom plate is under the gate valve, and the card box base is assembled with the card box base through a fixing member.
The multi-index detection microfluidic cartridge according to claim 1, wherein the piston pump includes a push-pull piston rod and a piston barrel to control the up and down movement of the piston rod according to a control signal from an external instrument.
The multi-index detection microfluidic cartridge of claim 1, wherein the number of the liquid storage tube is at least two.
The multi-index detection microfluidic cartridge according to claim 1, wherein a hydrophobic gas-permeable membrane is provided at the position of the vent hole of the liquid storage tube cover.
The multi-index detection microfluidic cartridge according to claim 1, wherein the number of the multi-index detection reaction chamber of the cartridge base is at least one.
The multi-index detection microfluidic cartridge according to claim 1, wherein the bottom of the multi-index detection reaction chamber of the cartridge base is provided with a notch for fixing a flat base.
The multi-index detection microfluidic cartridge according to claim 1, wherein the cover plate is made of a transparent material to obtain a biological detection response.
The multi-index detection microfluidic cartridge according to claim 1, wherein the sealing gasket and the gate valve placement groove of the cartridge base have the same shape and size, and are located at the through hole position of the cartridge base A through hole of the card box base is provided.
The multi-index detection microfluidic cartridge according to claim 1, wherein the base of the cartridge is made of transparent plastic, and the sealing gasket is made of elastic polymer material.
An application method for a multi-index detection microfluidic cartridge, characterized in that the multi-index detection microfluidic cartridge according to any one of claims 1-9 is used, wherein the method comprises:

Step S1, the reagents required for the reaction are stored in the liquid storage tube of the cartridge in advance, and after the user adds the sample into the liquid storage tube, the cartridge is placed in the supporting control instrument;

Step S2, using an external control instrument, through the cooperation control of the piston pump and the gate valve, to transfer, mix, and distribute the multi-step fluid, and control the target temperature and magnetic beads according to the detection reaction to complete The required biological test response;

Step S3, controlling the external control instrument to read the reaction result of the multi-index reaction chamber of the cartridge.