Microfluidic biochip integrating nucleic acid amplification and hybridization

Abstract

The application discloses a microfluidic biochip integrating nucleic acid amplification and hybridization, the microfluidic biochip comprises a substrate, the substrate is provided with an amplification chamber and a reaction chamber which are communicated with each other, the substrate is also provided with a driving module and a sample filling flow channel, and the driving module and the sample filling flow channel are respectively communicated with the amplification chamber; a plurality of reagent bins are further arranged on the substrate, are mutually independent and are respectively communicated with the reaction bin; during use, after a sample is injected into the amplification bin and amplified, the sample is controlled by the driving module to move into the reaction bin, different reagents required by a test are packaged in the reagent bins, the reagents enter the reaction bin to react with the sample, and finally a test result is obtained by observing the reaction bin. The reaction bin and the amplification bin are integrated on the same substrate, so that on one hand, the equipment integration degree is improved; on the other hand, amplification and reaction of the sample can be achieved through one device, frequent transfer of the amplified sample is avoided, and the test efficiency is improved.

Description

Technical Field

[0001] The present application relates to the technical field of heat dissipation equipment, and in particular to a microfluidic biochip integrating nucleic acid amplification and hybridization. Background Art

[0002] Currently, microfluidic biochips with nucleic acid amplification (PCR) and nucleic acid hybridization are widely used for rapid detection in the fields of medicine, agriculture, etc.

[0003] In the prior art, nucleic acid amplification (PCR) and nucleic acid hybridization are completed on two different modules respectively. However, some experiments require nucleic acid amplification (PCR) and nucleic acid hybridization to be performed successively, which results in the need to transfer samples between different modules. This not only involves certain experimental risks, but also has low detection efficiency. Utility Model Content

[0004] The main purpose of the present application is to provide a microfluidic biochip integrating nucleic acid amplification and hybridization, aiming to solve the defect of low detection efficiency existing in the prior art.

[0005] This application achieves the above objectives through the following technical solutions:

[0006] A microfluidic biochip integrating nucleic acid amplification and hybridization, comprising a substrate;

[0007] A driving module, wherein the driving module is disposed on the substrate;

[0008] an amplification chamber, wherein the amplification chamber is disposed on the substrate, and the driving module is connected to the amplification chamber; a sample filling channel connected to the amplification chamber is also disposed on the substrate;

[0009] A reaction chamber, wherein the reaction chamber is disposed on the substrate and is connected to the amplification chamber;

[0010] A plurality of reagent chambers, each of which is disposed on the substrate, each of which is independent of each other, and each of which is connected to the reaction chamber respectively.

[0011] Optionally, the driving module includes an airbag, which is connected to the amplification chamber through a first microchannel; and a first membrane pump for controlling the on-off state of the first microchannel is provided on the first microchannel.

[0012] Optionally, the sample injection channel is connected to the first microchannel; and a second membrane pump for controlling the on-off state of the sample injection channel is also provided on the sample injection channel.

[0013] Optionally, a heat conductive sheet is embedded in the bottom surface of the substrate, and the heat conductive sheet is arranged on the lower side of the amplification chamber; and a packaging film is arranged on the top of the amplification chamber.

[0014] Optionally, the amplification chamber is connected to the reaction chamber via a second microfluidic channel, and a third membrane pump for controlling the on / off state of the second microfluidic channel is provided on the second microfluidic channel.

[0015] Optionally, a membrane chip for hybridization reaction is arranged in the reaction chamber; a drainage pipe is arranged at the bottom of the reaction chamber, and an exhaust pipe is also arranged on the reaction chamber.

[0016] Optionally, the reagent chamber includes a chamber body disposed on the substrate, each of the chamber bodies being provided with a packaging film; and a liquid capsule for encapsulating the reagent is disposed in the chamber body.

[0017] Optionally, a ejector pin is further provided at the bottom of the bin body, and the ejector pin contacts the bottom of the liquid capsule.

[0018] Optionally, the chamber body is connected to an infusion channel, and the infusion channel is connected to the reaction chamber.

[0019] Optionally, a plurality of freeze-dried reagent chambers are further provided on the substrate, some of the reagent chambers correspond to each of the freeze-dried reagent chambers, and the corresponding reagent chambers and freeze-dried reagent chambers are connected in series with the reaction chamber through an infusion channel.

[0020] Compared with the prior art, this application has the following beneficial effects:

[0021] The present application includes a substrate, on which an amplification chamber and a reaction chamber that are interconnected are arranged, and a driving module and a sample injection channel are also arranged on the substrate, and the driving module and the sample injection channel are respectively connected to the amplification chamber; the substrate is also provided with a plurality of reagent chambers, each of which is independent of each other, and each of which is respectively connected to the reaction chamber.

[0022] When the biochip described in the present application is used, the sample is connected to an external filling device through the sample filling channel, so that the sample is filled into the amplification chamber. After the sample amplification is completed, the amplified sample is controlled by the driving module to move into the reaction chamber. Different reagents required for the test are encapsulated in each reagent chamber. Reagents are selected according to actual needs, and each reagent is transported to the reaction chamber to react with the sample. Finally, the test result is obtained by observing the reaction chamber.

[0023] Compared with the prior art, the present application integrates the reaction chamber and the amplification chamber on the same substrate, which improves the device integration on the one hand; on the other hand, the amplification and reaction of the sample can be achieved through one device, avoiding the frequent transfer of the amplified sample, which is conducive to improving the test efficiency;

[0024] At the same time, since there is no need to transfer the amplified samples between different devices, the subsequent reaction can be carried out as soon as the amplification is completed, which not only helps to improve work efficiency, but also can effectively ensure the activity of each material in the subsequent reaction, which is conducive to improving the accuracy of the reaction.

[0025] Secondly, the device described in the present application has both amplification and reaction functions, and the product has more abundant functions and can meet various usage requirements. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 A schematic diagram of the structure of a microfluidic biochip integrating nucleic acid amplification and hybridization provided in Embodiment 1 of the present application;

[0027] Figure 2 A side view of a microfluidic biochip integrating nucleic acid amplification and hybridization provided in embodiment 1 of the present application;

[0028] Figure 3 for Figure 1 Sectional view along line AA;

[0029] Figure markings: 1-substrate, 2-amplification chamber, 3-sample filling channel, 4-reaction chamber, 5-reagent chamber, 6-air bag, 7-first microchannel, 8-first membrane pump, 9-second membrane pump, 10-heat conductive sheet, 11-packaging membrane, 12-second microchannel, 13-third membrane pump, 14-drain pipe, 15-exhaust pipe, 16-liquid bag, 17-thrust needle, 18-infusion channel, 19-lyophilized reagent chamber, 501-chamber body, 502-packaging film.

[0030] The purpose, features and advantages of this application will be further described in conjunction with embodiments and with reference to the accompanying drawings. DETAILED DESCRIPTION

[0031] The following will be combined with the drawings in the embodiments of the utility model to clearly and completely describe the technical solutions in the embodiments of the utility model. Obviously, the described embodiments are only part of the embodiments of the utility model, not all of the embodiments. Based on the embodiments of the utility model, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the utility model.

[0032] It should be noted that all directional indications in the embodiments of the present invention (such as up, down, left, right, front, back...) are only used to explain the relative position relationship, movement status, etc. between the components under a certain specific posture (as shown in the accompanying drawings). If the specific posture changes, the directional indication will also change accordingly.

[0033] In the present invention, unless otherwise clearly specified and limited, the terms "connection", "fixation", etc. should be understood in a broad sense. For example, "fixation" can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements, unless otherwise clearly defined. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0034] In addition, if there are descriptions involving "first", "second", etc. in the embodiments of the utility model, the descriptions of "first", "second", etc. are only used for descriptive purposes and cannot be understood as indicating or implying their relative importance or implicitly indicating the number of technical features indicated. Therefore, the features defined as "first" and "second" may explicitly or implicitly include at least one of the features. In addition, the meaning of "and / or" appearing in the full text includes three parallel schemes. Taking "A and / or B" as an example, it includes scheme A, or scheme B, or a scheme that satisfies both A and B. In addition, the technical solutions between the various embodiments can be combined with each other, but it must be based on the ability of ordinary technicians in this field to implement. When the combination of technical solutions is contradictory or cannot be implemented, it should be deemed that such a combination of technical solutions does not exist and is not within the scope of protection required by the utility model.

[0035] Implementation Method 1

[0036] Reference Figures 1 to 3 This embodiment is an optional embodiment of the present application, which discloses a microfluidic biochip integrating nucleic acid amplification and hybridization, including a substrate 1, which is a flat plate, on which an amplification chamber 2 and a reaction chamber 4 are arranged, and a driving module is also arranged, the driving module includes an air bag 6, and the air bag 6 is arranged on the substrate 1, and a first microchannel 7 is also arranged on the substrate 1, the first microchannel 7 connects the outlet end of the air bag 6 with the inlet end of the amplification chamber 2, and a first membrane pump 8 for controlling its on and off state is arranged on the first microchannel 7.

[0037] The substrate 1 is also provided with a sample injection channel 3, one end of which is connected in parallel with the first microchannel 7, and the other end thereof is adapted to an external injection device, and the sample injection channel 3 is also provided with a second membrane pump 9 for controlling its on and off state;

[0038] The substrate 1 is also provided with a second microchannel 12, and the two ends of the second microchannel 12 are respectively connected to the outlet end of the amplification chamber 2 and an inlet end of the reaction chamber 4; the second microchannel 12 is provided with a third membrane pump 13 for controlling its on-off state;

[0039] Meanwhile, a heat conducting sheet 10 is embedded in the substrate 1. The heat conducting sheet 10 is made of a good heat conductor such as copper or aluminum. The heat conducting sheet 10 is arranged at the lower side of the amplification chamber 2. A packaging film 11 is arranged on the top of the amplification chamber 2, and the amplification chamber 2 is sealed by the packaging film 11.

[0040] When in use, the heat conductive sheet 10 is closely attached to the external heating assembly, so that the temperature of the amplification chamber 2 can be quickly adjusted and controlled by external heating to meet the temperature requirements of different amplification tests;

[0041] At the same time, compared with the microfluidic chip with a lid or the microfluidic chip with a PCR tube, the present application can increase the heating and cooling rate through the packaging film 11. Actual measurement shows that the average temperature rising and falling rate of the present application is 5°C per second, and the maximum rate is 10°C, which enhances the PCR amplification reaction rate.

[0042] The reaction chamber 4 is provided with a membrane chip for hybridization reaction; a drain pipe 14 is provided at the bottom of the reaction chamber 4, and an exhaust pipe 15 is provided on the reaction chamber 4; various waste liquids in the reaction chamber 4 can be conveniently discharged through the drain pipe 14, and waste gas generated during the reaction process can be discharged in time through the exhaust pipe 15; the drain pipe 14 and the exhaust pipe 15 are both provided with regulating valves for controlling their on-off states;

[0043] A plurality of reagent chambers 5 are also provided on the substrate 1. The structures of the reagent chambers 5 are completely the same. Each reagent chamber 5 comprises a chamber body 501 provided on the substrate 1. The chamber body 501 is provided with an opening at the top and is closed by a packaging clip. At least one ejector pin 17 is provided on the bottom surface of each chamber body 501. A liquid capsule 16 is also provided in each chamber body 501.

[0044] The liquid capsule 16 is made of aluminum foil, and the thickness of the top of the liquid capsule 16 is greater than the thickness of the bottom. The bottom of the liquid capsule 16 is in contact with the ejector pin 17. A quantitative reagent for detection is encapsulated in the liquid capsule 16.

[0045] Each of the reagent chambers 5 is provided with an infusion channel 18, the inlet end of the infusion channel 18 is connected to the reagent chamber 5, and the outlet end thereof is connected to the reaction chamber 4;

[0046] Furthermore, a plurality of freeze-dried reagent chambers 19 are provided on the substrate 1. It should be noted that the number of freeze-dried reagent chambers 19 is flexibly determined according to needs; each of the freeze-dried reagent chambers 19 is connected to an infusion channel 18 respectively. If two freeze-dried reagent chambers 19 are provided, there must be two reagent chambers 5 corresponding to the two freeze-dried reagent chambers 19 respectively. In the freeze-dried reagent chambers 19 and reagent chambers 5 corresponding to each other, the outlet end of the reagent chamber 5 is connected to the inlet end of the freeze-dried reagent chamber 19 through the infusion channel 18, and the outlet end of the freeze-dried reagent chamber 19 is connected to the reaction chamber 4 through the infusion channel 18, that is, the freeze-dried reagent chambers 19 and reagent chambers 5 corresponding to each other are connected in series through the infusion channel 18 and then connected to the reagent chamber 5.

[0047] When in use, the freeze-dried reagent compartment 19 is used to store freeze-dried reagents to meet the storage requirements of different types of reagents; if no freeze-dried reagent is involved in the experiment, the freeze-dried reagent compartment 19 is empty.

[0048] Accordingly, the present application also discloses a method for using the above-mentioned microfluidic biochip, comprising the following steps:

[0049] S1, first open the second membrane pump, and at the same time close the first membrane pump and the third membrane pump, and inject the sample from the sample filling channel into the amplification chamber through an external filling device (such as a pipette); then close the second membrane pump;

[0050] S2, connecting the microfluidic control chip filled with samples to an external device; and turning on the external device at the same time;

[0051] S3. The external device controls the temperature of the amplification chamber by heating, thereby completing the amplification of the sample; it should be noted that the above amplification process is automatically controlled by the program, and parameters such as amplification temperature and amplification time can be preset by the staff;

[0052] S4. After the amplification is completed, the first membrane pump and the third membrane pump are turned on, and the air bag is squeezed manually or by external equipment, so as to force the sample in the amplification chamber to enter the reaction chamber until the number of samples in the reaction chamber meets the requirement;

[0053] S5. According to the actual needs of the test, one or more reagent chambers are selected to be actually input into the reaction chamber; it should be noted that the order in which each reagent is input into the reaction chamber, the input amount, and whether to input simultaneously can be flexibly adjusted by the staff according to actual needs;

[0054] The force between the liquid capsule and the ejector pin is increased by squeezing the encapsulating film, so that the liquid capsule is punctured by the ejector pin. After the liquid capsule is punctured, the liquid actually flows out and enters the reaction chamber through the infusion channel under squeezing;

[0055] If there is a freeze-dried reagent, the liquid reagent will be mixed with the freeze-dried reagent in the freeze-dried reagent chamber before entering the reaction chamber;

[0056] S6, the membrane chip in the chamber is immersed in the reagent liquid. During the reaction, the external device vibrates the reagent liquid in the chamber. The waste liquid generated after each hybridization reaction process is discharged through the drain pipe at the bottom of the reaction chamber;

[0057] After the reaction is completed, the color development results are displayed on the membrane chip, and external equipment can take pictures, save, identify and interpret through the window. The results can also be observed and judged by human eyes.

[0058] Compared with the prior art, the present application integrates the reaction chamber and the amplification chamber on the same substrate, which improves the device integration on the one hand; on the other hand, the amplification and reaction of the sample can be achieved through one device, avoiding the frequent transfer of the amplified sample, which is conducive to improving the test efficiency;

[0059] At the same time, since there is no need to transfer the amplified samples between different devices, the subsequent reaction can be carried out as soon as the amplification is completed, which not only helps to improve work efficiency, but also can effectively ensure the activity of each material in the subsequent reaction, which is conducive to improving the accuracy of the reaction.

[0060] Secondly, the device described in the present application has both amplification and reaction functions, and the product has more abundant functions and can meet various usage requirements.

[0061] The above are only preferred embodiments of the present application, and are not intended to limit the patent scope of the present application. Any equivalent structure or equivalent process transformation made using the contents of the present application specification and drawings, or directly or indirectly applied in other related technical fields, are also included in the patent protection scope of the present application.

Claims

1. A microfluidic biochip integrating nucleic acid amplification and hybridization, characterized in that: comprising a substrate (1); A driving module, wherein the driving module is arranged on the substrate (1); an amplification chamber (2), the amplification chamber (2) being arranged on the substrate (1), the driving module being in communication with the amplification chamber (2); and a sample injection channel (3) being in communication with the amplification chamber (2) being further arranged on the substrate (1); A reaction chamber (4), wherein the reaction chamber (4) is disposed on the substrate (1), and the reaction chamber (4) is connected to the amplification chamber (2); A plurality of reagent chambers (5), each of which is arranged on the substrate (1), each of which is independent of each other, and each of which is connected to the reaction chamber (4).

2. The microfluidic biochip integrating nucleic acid amplification and hybridization according to claim 1, characterized in that: The driving module comprises an air bag (6), and the air bag (6) is connected to the amplification chamber (2) through a first microfluidic channel (7); the first microfluidic channel (7) is provided with a first membrane pump (8) for controlling its on and off state.

3. The microfluidic biochip integrating nucleic acid amplification and hybridization according to claim 2, characterized in that: The sample injection flow channel (3) is in communication with the first micro-flow channel (7); the sample injection flow channel (3) is also provided with a second membrane pump (9) for controlling its on / off state.

4. The microfluidic biochip integrating nucleic acid amplification and hybridization according to claim 1, characterized in that: A heat conducting sheet (10) is also embedded in the bottom surface of the substrate (1), and the heat conducting sheet (10) is arranged on the lower side of the amplification chamber (2); a packaging film (11) is arranged on the top of the amplification chamber (2).

5. A microfluidic biochip integrating nucleic acid amplification and hybridization according to claim 1 or 4, characterized in that: The amplification chamber (2) is connected to the reaction chamber (4) via a second microfluidic channel (12), and a third membrane pump (13) is provided on the second microfluidic channel (12) for controlling its on / off state.

6. The microfluidic biochip integrating nucleic acid amplification and hybridization according to claim 1, characterized in that: A membrane chip for hybridization reaction is arranged in the reaction chamber (4); a liquid discharge pipe (14) is arranged at the bottom of the reaction chamber (4); and an exhaust pipe (15) is also arranged on the reaction chamber (4).

7. The microfluidic biochip integrating nucleic acid amplification and hybridization according to claim 1, characterized in that: The reagent chamber (5) comprises a chamber body (501) arranged on the substrate (1), and each chamber body (501) is provided with a packaging film (502); and a liquid capsule (16) for packaging the reagent is provided in the chamber body (501).

8. The microfluidic biochip integrating nucleic acid amplification and hybridization according to claim 7, characterized in that: The bottom of the storage body (501) is also provided with an ejector pin (17), and the ejector pin (17) is in contact with the bottom of the liquid capsule (16).

9. The microfluidic biochip integrating nucleic acid amplification and hybridization according to claim 7, characterized in that: The chamber body (501) is connected to a liquid infusion channel (18), and the liquid infusion channel (18) is connected to the reaction chamber (4).

10. The microfluidic biochip integrating nucleic acid amplification and hybridization according to claim 9, characterized in that: The substrate (1) is also provided with a plurality of freeze-dried reagent chambers (19), and some of the reagent chambers (5) correspond to the freeze-dried reagent chambers (19). The corresponding reagent chambers (5) and freeze-dried reagent chambers (19) are connected in series via an infusion channel (18) and then connected to the reaction chamber (4).

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