Microflow pump and application and use method thereof

Abstract

The invention relates to the field of micro-flow pumps and the field of biomedicine, and discloses a micro-flow pump and application and using method.The micro-flow pump comprises at least two micro-flow sub-pumps, each micro-flow sub-pump comprises a liquid containing cavity, at least one side wall of each liquid containing cavity is a piezoelectric ceramic piece, and the piezoelectric ceramic pieces are arranged on the side walls of the liquid containing cavities; the piezoelectric ceramic piece vibrates to enable the liquid containing cavity to be in a contraction or expansion state; the liquid flowing-in channel is communicated with the liquid containing cavity through a first one-way valve; the liquid outflow channel is communicated with the liquid containing cavity through a second one-way valve; and the isothermal reaction chamber is communicated with the liquid outflow channel and is used for carrying out a nucleic acid isothermal amplification reaction. The micro-flow pump has the advantages that the quantitative input of reactants is realized, the reaction efficiency is maximized, the economic cost is reduced, simplicity, convenience and high efficiency are realized, and the detection and research in the aspect of a nucleic acid isothermal amplification technology are facilitated.

Description

Technical Field

[0001] The invention relates to the field of microfluidic pumps and biomedicine, and in particular to a microfluidic pump and an application and use method thereof. Background Art

[0002] Isothermal nucleic acid amplification technology, which is to expand the number of copies of specific DNA or RNA fragments at a specific temperature, is widely used in clinical diagnosis due to its rapid, sensitive and specific advantages. In the biomedical research process such as nucleic acid detection and vaccine development, the directional and quantitative delivery of trace amounts of liquid runs through the entire process of chemical analysis, drug delivery, molecular recognition, etc. In addition, modern molecular detection technology has the demand for speed and simplicity, requiring simplified and efficient equipment. However, traditional nucleic acid amplification detection relies on the operation of professional technicians, and inaccurate control of the reactant quantity can easily cause waste, time-consuming and labor-intensive. Summary of the invention

[0003] The purpose of the present invention is to overcome the problems of inaccurate and time-consuming control of the amount of nucleic acid amplification detection reactants in the prior art, and to provide a microfluidic pump and its application and use method. The microfluidic pump has the function of realizing quantitative input of reactants, maximizing the efficiency of the reaction, and reducing economic costs. It is simple and efficient, and is convenient for the detection and research of nucleic acid isothermal amplification technology.

[0004] In order to achieve the above object, the present invention provides a microfluidic pump for isothermal nucleic acid amplification reaction, wherein the microfluidic pump comprises at least two microfluidic pumps, and the microfluidic pump comprises:

[0005] A liquid containing chamber, wherein at least one side wall of the liquid containing chamber is a piezoelectric ceramic sheet, and the piezoelectric ceramic sheet vibrates to put the liquid containing chamber into a contracted or expanded state;

[0006] The liquid flows into the channel and communicates with the liquid containing chamber through the first one-way valve;

[0007] a liquid outflow channel, connected to the liquid containing chamber via a second one-way valve;

[0008] An isothermal reaction chamber, the isothermal reaction chamber being in communication with the liquid outflow channel and being used for performing isothermal nucleic acid amplification reaction;

[0009] When the liquid containing chamber is in an expanded state, the first one-way valve is opened and the second one-way valve is closed, and the liquid flows from the first one-way valve into the liquid containing chamber;

[0010] When the liquid containing chamber is in a contracted state, the first one-way valve is closed and the second one-way valve is opened, and the liquid in the liquid containing chamber flows out from the second one-way valve and flows into the isothermal reaction chamber through the liquid outflow channel.

[0011] Preferably, the microfluidic pump comprises an injection molded structure, a liquid containing cavity is formed between the piezoelectric ceramic sheet and the injection molded structure, the injection molded structure comprises a channel layer and a buffer cavity layer, and the channel layer is provided with a liquid inlet channel and a liquid outlet channel.

[0012] Preferably, the microfluidic pump further comprises a driving circuit, wherein the driving circuit is connected to the piezoelectric ceramic sheet and is used to apply a voltage to the piezoelectric ceramic sheet.

[0013] Preferably, the microfluidic pump further comprises a sealing layer, and the sealing layer covers the piezoelectric ceramic sheet.

[0014] Preferably, a temperature control device and a temperature measuring device are provided in the isothermal reaction chamber.

[0015] Further preferably, the temperature control device is used to control the temperature in the isothermal reaction chamber.

[0016] Further preferably, the temperature measuring device is used to detect the temperature in the isothermal reaction chamber.

[0017] Preferably, when the microfluidic pump has two microfluidic pumps, the two microfluidic pumps may be independent of each other, or the two microfluidic pumps may share a liquid containing cavity and a piezoelectric ceramic plate.

[0018] A second aspect of the present invention provides an application of the above-mentioned microfluidic pump in a nucleic acid isothermal amplification reaction.

[0019] A third aspect of the present invention provides a method for using the microfluidic pump, comprising the following steps:

[0020] S1, turning on the driving circuit to make the piezoelectric ceramic sheet vibrate so that the liquid containing cavity is in a contracted or expanded state;

[0021] S2. When the liquid receiving chamber of one of the microfluidic pumps is in an expanded state, the first one-way valve of the microfluidic pump is opened, and the second one-way valve is closed, and the nucleic acid solution flows into the liquid receiving chamber of the microfluidic pump through the liquid inflow channel of the microfluidic pump and flows into the liquid receiving chamber of the microfluidic pump through the first one-way valve. When the liquid receiving chamber of the microfluidic pump is in a contracted state, the first one-way valve is closed, and the second one-way valve is opened, and the nucleic acid solution in the liquid receiving chamber flows out from the second one-way valve and flows into the isothermal reaction chamber through the liquid outflow channel.

[0022] S3, when the liquid receiving chamber of another microfluidic pump is in an expanded state, the first one-way valve of the microfluidic pump is opened, and the second one-way valve is closed, the enzyme solution flows in from the liquid inflow channel of the microfluidic pump, and flows into the liquid receiving chamber of the microfluidic pump through the first one-way valve; when the liquid receiving chamber of the microfluidic pump is in a contracted state, the first one-way valve is closed, and the second one-way valve is opened, the enzyme solution in the liquid receiving chamber flows out from the second one-way valve, and flows into the isothermal reaction chamber through the liquid outflow channel;

[0023] S4. The nucleic acid solution and the enzyme solution are contacted in the isothermal reaction chamber to perform a nucleic acid isothermal amplification reaction.

[0024] The present invention provides a miniaturized and integrated microfluidic pump for isothermal nucleic acid amplification reaction. Through the microfluidic pump of the present invention, the precise quantitative input of nucleic acid and other reaction substances can be achieved by controlling the vibration of the piezoelectric ceramic sheet, and the isothermal amplification reaction is carried out after mixing in the isothermal reaction chamber. The microfluidic pump has a simple structure and can meet the requirements of modern molecular detection technology for rapid and simple operation. It has great potential in the detection and research of nucleic acid isothermal amplification technology and has great practical application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 is a schematic structural diagram of the microfluidic pump in Example 1 of the present invention;

[0026] Figure 2 is a schematic structural diagram of the microfluidic pump in Example 1 of the present invention;

[0027] Figure 3 It is a schematic diagram of the structure of the microfluidic pump in Example 2 of the present invention.

[0028] Description of Reference Numerals

[0029] 1. Liquid containing chamber; 2. Piezoelectric ceramic sheet; 3. Liquid inflow channel; 4. First one-way valve; 5. Liquid outflow channel; 6. Second one-way valve; 7. Isothermal reaction chamber. DETAILED DESCRIPTION

[0030] The specific implementation of the present invention is described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to illustrate and explain the present invention, and are not used to limit the present invention.

[0031] The endpoints and any values ​​of the ranges disclosed in this article are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of each range, the endpoint values ​​of each range and the individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed in this article.

[0032] In addition, if there are descriptions involving "first", "second", etc. in the embodiments of the present invention, the descriptions of "first", "second", etc. are only used for descriptive purposes and cannot be understood as indicating or suggesting their relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include at least one of the features. In addition, the technical solutions between the various embodiments provided by the present invention can be combined with each other, but they must be based on the ability of ordinary technicians in the field to implement them. When the combination of technical solutions is contradictory or cannot be implemented, it should be deemed that such combination of technical solutions does not exist and is not within the scope of protection required by the present invention.

[0033] Example 1

[0034] like Figure 1 The microfluidic pump for isothermal nucleic acid amplification reaction shown in the figure comprises at least two microfluidic pumps, and the microfluidic pump comprises:

[0035] A liquid containing chamber 1, wherein at least one side wall of the liquid containing chamber 1 is a piezoelectric ceramic sheet 2, and the piezoelectric ceramic sheet 2 vibrates to put the liquid containing chamber 1 into a contraction or expansion state;

[0036] The liquid flows into the channel 3, and the liquid flows into the channel 3 and communicates with the liquid containing chamber 1 through the first one-way valve 4;

[0037] A liquid outflow channel 5, the liquid outflow channel 5 is connected to the liquid containing chamber 1 through a second one-way valve 6;

[0038] An isothermal reaction chamber 7, which is connected to the liquid outflow channel 5 and is used for performing isothermal nucleic acid amplification reaction;

[0039] When the liquid containing chamber 1 is in an expanded state (at this time, the air pressure in the liquid containing chamber 1 is lower than the external atmospheric pressure), the first one-way valve 4 is opened, and the second one-way valve 6 is closed, and the liquid flows from the first one-way valve 4 into the liquid containing chamber 1;

[0040] When the liquid holding chamber 1 is in a contracted state (at this time the air pressure in the liquid holding chamber 1 is higher than the external atmospheric pressure), the first one-way valve 4 is closed and the second one-way valve 6 is opened, and the liquid in the liquid holding chamber 1 flows out from the second one-way valve 6 and flows into the isothermal reaction chamber 7 through the liquid outflow channel 5.

[0041] The first one-way valve 4 and the second one-way valve 6 are both used for one-way inflow and outflow of liquid; the first one-way valve 4 and the second one-way valve can be used as follows Figure 1 The umbrella-shaped one-way valve shown in FIG. Figure 2 The cantilever valve shown has the effect of preventing liquid backflow, etc.

[0042] The microfluidic pump of the present invention further comprises a driving circuit, which is connected to the piezoelectric ceramic sheet 2 and is used to apply voltage to the piezoelectric ceramic sheet 2. The voltage signal provided by the driving circuit is an alternating signal.

[0043] Specifically, when voltage is applied to the piezoelectric ceramic piece 2, the piezoelectric ceramic piece 2 as a driving source will vibrate up and down. When the piezoelectric ceramic piece 2 vibrates in a direction outside the liquid holding chamber 1, a low pressure is formed in the liquid holding chamber 1, and the pressure in the liquid holding chamber 1 is less than the external atmospheric pressure. The liquid holding chamber 1 is in an expanded state. At this time, the first one-way valve 4 is opened, and the second one-way valve 6 is closed, and the liquid flows into the liquid holding chamber 1 from the first one-way valve 4; when the piezoelectric ceramic piece 2 vibrates in a direction inside the liquid holding chamber 1 under the drive of voltage, a high pressure is formed in the liquid holding chamber 1, and the pressure in the liquid holding chamber 1 is greater than the external atmospheric pressure, so that the liquid holding chamber 1 is in a contracted state. At this time, the first one-way valve 4 is closed, and the second one-way valve 6 is opened, and the liquid in the liquid holding chamber 1 flows out from the second one-way valve 6, flows into the isothermal reaction chamber 7 through the liquid outflow channel 5, and performs a nucleic acid isothermal amplification reaction.

[0044] The specific sizes of the piezoelectric ceramic sheet 2, the liquid containing cavity 1, the liquid inlet channel 3, the liquid outlet channel 5 and the isothermal reaction chamber 7 are determined according to actual needs.

[0045] Furthermore, the expansion state of the liquid holding chamber 1 is the state in which the pressure inside the liquid holding chamber 1 is less than the external atmospheric pressure; the contraction state of the liquid holding chamber 1 refers to the state in which the pressure inside the liquid holding chamber 1 is greater than the external atmospheric pressure.

[0046] Among them, the piezoelectric ceramic sheet 2 has a high sensitivity and can be made very thin, so the overall size of the microfluidic pump can be reduced. If an AC square wave signal is applied to the piezoelectric ceramic sheet 2, the above process can be repeated, and different flow rates can be achieved by adjusting the amplitude and frequency of the signal voltage to achieve the purpose of accurately quantitatively inputting the reactants.

[0047] The microfluidic pump of the present invention controls the contraction or expansion state of the liquid holding chamber 1 by deforming the piezoelectric ceramic sheet 2 under the drive of voltage, so that the liquid reaction substance passes through the liquid holding chamber 1 from the liquid inflow channel 3, flows out from the liquid outflow channel 5, enters the isothermal reaction chamber 7, and undergoes nucleic acid isothermal amplification reaction after mixing. The relationship between the pressure of the liquid holding chamber 1 and the external atmospheric pressure can be adjusted by adjusting the voltage amplitude and frequency, thereby controlling the liquid reaction substance to flow in from the liquid inflow channel 3, flow out from the liquid outflow channel 5, and control the flow rate of the liquid reaction substance. Moreover, since the piezoelectric ceramic sheet 2 has a high sensitivity, it can be made into a small and thin sheet to make a miniaturized microfluidic pump, which has great potential in the field of biomedicine.

[0048] Furthermore, the microfluidic pump of the present invention includes an injection molded structure, a liquid containing cavity 1 is formed between the piezoelectric ceramic sheet 2 and the injection molded structure, the injection molded structure includes a channel layer and a buffer cavity layer, and the channel layer is provided with a liquid inlet channel 3 and a liquid outlet channel 5.

[0049] Among them, the purpose of setting the buffer chamber layer is to temporarily store the liquid reactant flowing in from the first one-way valve 4, so as to ensure that the source of liquid reactant in the liquid holding chamber 1 is sufficient, and by controlling the opening degree of the second one-way valve 6, the liquid reactant can be controlled to flow out of the liquid channel 5 to control the liquid flow rate.

[0050] In a specific embodiment, the material and size of the injection molded part structure depend on the specific application scenario (for example, for drugs, it is necessary to consider whether it will cause contamination to the drug, and for biological experiments, whether it is biocompatible).

[0051] Preferably, the inflow and outflow flow of the liquid can be controlled by controlling the switching amplitude of the first one-way valve 4 and the second one-way valve 6. The switching amplitude of the first one-way valve 4 and the second one-way valve 6 is determined by the voltage applied to the piezoelectric ceramic sheet 2. The amplitude of the piezoelectric ceramic sheet 2 can be further adjusted by changing the voltage applied to the piezoelectric ceramic sheet 2 by the driving circuit to accurately control the flow. The greater the voltage applied to the piezoelectric ceramic sheet 2, the greater the pressure difference between the liquid holding chamber 1 and the external atmospheric pressure, the greater the switching amplitude of the first one-way valve 4 and the second one-way valve 6, and therefore the greater the inflow and outflow flow of the liquid.

[0052] The microfluidic pump in the present invention further comprises a sealing layer, which covers the piezoelectric ceramic sheet 2 and seals the piezoelectric ceramic sheet 2 to prevent the piezoelectric ceramic sheet 2 from being exposed to humid air for a long time and causing short circuits, device damage, etc.

[0053] Furthermore, a temperature control device and a temperature measuring device are also provided in the isothermal reaction chamber 7, and the isothermal reaction chamber 7 is used to control the progress of the nucleic acid isothermal amplification reaction; wherein the temperature control device is used to control the temperature in the isothermal reaction chamber 7 to achieve heating and maintain a constant temperature, and the temperature measuring device is used to detect the temperature in the isothermal reaction chamber 7 and provide real-time feedback of the current temperature.

[0054] The isothermal reaction chamber 7 can adjust the temperature control device therein according to the actual application conditions of the nucleic acid isothermal amplification reaction, thereby controlling the temperature change and maintaining a constant temperature to provide a place for the reaction.

[0055] In this embodiment, the microfluidic pump has two microfluidic pumps, such as Figure 1As shown, during actual use, one of the microfluidic pumps is used to control the inflow and outflow of nucleic acids, and the other microfluidic pump is used to control the inflow and outflow of enzymes required for the reaction. The nucleic acids and the enzymes required for the reaction eventually flow into the isothermal reaction chamber 7, and the nucleic acid isothermal amplification reaction is carried out in the isothermal reaction chamber 7.

[0056] Furthermore, in actual use, the nucleic acid and enzyme need to be prepared into corresponding nucleic acid solutions and enzyme solutions respectively, and then the nucleic acid isothermal amplification reaction is carried out in the microfluidic pump of the present invention. The preparation method and concentration of the nucleic acid solution and enzyme solution depend on the specific usage.

[0057] Among them, the control of the two microfluidic pumps does not interfere with each other. The same voltage can be applied at the same time to achieve the synchronous inflow and outflow of two reactants. Opposite voltages (opposite amplitudes, same frequency) can be applied separately to achieve one microfluidic pump flowing in one reactant while the other microfluidic pump flows out another reactant. Different voltages can also be applied separately to control different flow rates.

[0058] In actual use, when there are other types of reactants in addition to nucleic acids and enzymes, the microfluidic pump can also be equipped with more microfluidic pumps to flow in and out other types of reactants (that is, each microfluidic pump is responsible for the inflow and outflow of one reactant), and finally all flow into the isothermal reaction chamber 7, in which the nucleic acid isothermal amplification reaction is carried out; the specific number of microfluidic pumps depends on the specific usage requirements.

[0059] The method for using the microfluidic pump comprises the following steps:

[0060] S1, turning on the driving circuit, adjusting the voltage, applying an alternating signal voltage to the two piezoelectric ceramic sheets 2, so that the two piezoelectric ceramic sheets 2 vibrate up and down so that the two liquid containing chambers 1 are in a contracted or expanded state;

[0061] S2. Prepare a nucleic acid solution. When the liquid receiving chamber 1 of one of the microfluidic pumps is in an expanded state, the first one-way valve 4 of the microfluidic pump is opened, and the second one-way valve 6 is closed. The nucleic acid solution flows in from the liquid inflow channel 3 of the microfluidic pump, and flows into the liquid receiving chamber 1 of the microfluidic pump through the first one-way valve 4. When the liquid receiving chamber 1 of the microfluidic pump is in a contracted state, the first one-way valve 4 is closed, and the second one-way valve 6 is opened. The nucleic acid solution in the liquid receiving chamber 1 flows out from the second one-way valve 6, and flows into the isothermal reaction chamber 7 through the liquid outflow channel 5.

[0062] S3, preparing an enzyme solution. When the liquid receiving chamber 1 of another microfluidic pump is in an expanded state, the first one-way valve 4 of the microfluidic pump is opened, and the second one-way valve 6 is closed. The enzyme solution flows in from the liquid inflow channel 3 of the microfluidic pump, and flows into the liquid receiving chamber 1 of the microfluidic pump through the first one-way valve 4. When the liquid receiving chamber 1 of the microfluidic pump is in a contracted state, the first one-way valve 4 is closed, and the second one-way valve 6 is opened. The enzyme solution in the liquid receiving chamber 1 flows out from the second one-way valve 6, and flows into the isothermal reaction chamber 7 through the liquid outflow channel 5.

[0063] S4. The nucleic acid solution and the enzyme solution are contacted and mixed in the isothermal reaction chamber 7. The temperature control device in the isothermal reaction chamber 7 is adjusted to control the temperature in the isothermal reaction chamber 7. The temperature measuring device in the isothermal reaction chamber 7 detects the real-time temperature in the isothermal reaction chamber 7, provides suitable reaction conditions, and performs nucleic acid isothermal amplification reaction.

[0064] In the above-mentioned usage method, the nucleic acid isothermal amplification reaction is a conventional nucleic acid isothermal amplification reaction in the art, wherein the flow rates of the two microfluidic pumps, the types of nucleic acids and enzymes, and the ratios are determined according to the specific nucleic acid isothermal amplification reaction; if there are other types of reaction substances, a corresponding number of microfluidic pumps can be added according to the number of specific reaction substances; the time for applying the voltage and the magnitude of the applied voltage are determined according to the specific amount of reaction substances required.

[0065] Example 2

[0066] The microfluidic pump of this embodiment is the same as the microfluidic pump of embodiment 1, and has two microfluidic pumps. The difference is that the two microfluidic pumps share a liquid receiving chamber 1 and a piezoelectric ceramic plate 2. Figure 3 As shown, at this time, only one piezoelectric ceramic sheet 2 is used to realize that the liquid receiving chamber 1 of one microfluidic pump is in an expanded state while the liquid receiving chamber 1 of another microfluidic pump is in a contracted state. Under this structure, the flow rates of the two microfluidic pumps are the same;

[0067] The method of using the microfluidic pump in this embodiment is the same as that in Example 1, except that while the liquid holding chamber 1 of one microfluidic pump is in an expanded state, the liquid holding chamber 1 of the other microfluidic pump is in a contracted state. In this case, the flow rates of the nucleic acid solution and the enzyme solution are the same. This situation is only applicable to nucleic acid isothermal amplification reactions with only two reaction substances, nucleic acid and enzyme.

[0068] It should be understood that parts not elaborated in detail in this specification belong to the prior art.

[0069] The preferred embodiments of the present invention are described in detail above, but the present invention is not limited thereto. Within the technical concept of the present invention, the technical solution of the present invention can be subjected to a variety of simple modifications, including the combination of various technical features in any other suitable manner, and these simple modifications and combinations should also be regarded as the contents disclosed by the present invention and belong to the protection scope of the present invention.

Claims

1. A microfluidic pump for isothermal nucleic acid amplification reaction, characterized in that: The microfluidic pump comprises at least two microfluidic pumps, and the microfluidic pump comprises: A liquid containing chamber (1), wherein at least one side wall of the liquid containing chamber (1) is a piezoelectric ceramic sheet (2), and the piezoelectric ceramic sheet (2) vibrates to put the liquid containing chamber (1) into a contracted or expanded state; The liquid flows into the channel (3) and communicates with the liquid containing chamber (1) through the first one-way valve (4); A liquid outflow channel (5) is connected to the liquid containing chamber (1) via a second one-way valve (6); An isothermal reaction chamber (7), the isothermal reaction chamber (7) being in communication with the liquid outflow channel (5) and being used for performing a nucleic acid isothermal amplification reaction; When the liquid containing chamber (1) is in an expanded state, the first one-way valve (4) is opened and the second one-way valve (6) is closed, and the liquid flows from the first one-way valve (4) into the liquid containing chamber (1); When the liquid containing chamber (1) is in a contracted state, the first one-way valve (4) is closed and the second one-way valve (6) is opened, and the liquid in the liquid containing chamber (1) flows out from the second one-way valve (6) and flows into the isothermal reaction chamber (7) through the liquid outflow channel (5).

2. The microfluidic pump according to claim 1, characterized in that: The microfluidic pump comprises an injection molded structure, a liquid containing cavity (1) is formed between the piezoelectric ceramic sheet (2) and the injection molded structure, and the injection molded structure comprises a channel layer, wherein the channel layer is provided with a liquid inflow channel (3) and a liquid outflow channel (5).

3. The microfluidic pump according to claim 1, characterized in that: The microfluidic pump further comprises a driving circuit, which is connected to the piezoelectric ceramic sheet (2) and is used to apply a voltage to the piezoelectric ceramic sheet (2).

4. The microfluidic pump according to claim 1, characterized in that: The microfluidic pump further comprises a sealing layer, wherein the sealing layer covers the piezoelectric ceramic sheet (2).

5. The microfluidic pump according to claim 1, characterized in that: The isothermal reaction chamber (7) is provided with a temperature control device and a temperature measuring device.

6. The microfluidic pump according to claim 5, characterized in that: The temperature control device is used to control the temperature in the isothermal reaction chamber (7).

7. The microfluidic pump according to claim 5 or 6, characterized in that: The temperature measuring device is used to detect the temperature in the isothermal reaction chamber (7).

8. The microfluidic pump according to claim 1, characterized in that: When the microfluidic pump has two microfluidic sub-pumps, the two microfluidic sub-pumps share a liquid containing cavity (1) and a piezoelectric ceramic plate (2).

9. Use of the microfluidic pump according to any one of claims 1 to 8 in a nucleic acid isothermal amplification reaction.

10. The method for using the microfluidic pump according to any one of claims 1 to 8, characterized in that: The following steps are involved: S1, turning on the driving circuit to make the piezoelectric ceramic sheet (2) vibrate so that the liquid containing chamber (1) is in a contracted or expanded state; S2. When the liquid containing chamber (1) of one of the microfluidic pumps is in an expanded state, the first one-way valve (4) of the microfluidic pump is opened, and the second one-way valve (6) is closed, and the nucleic acid solution flows in from the liquid inflow channel (3) of the microfluidic pump, and flows into the liquid containing chamber (1) of the microfluidic pump through the first one-way valve (4); when the liquid containing chamber (1) of the microfluidic pump is in a contracted state, the first one-way valve (4) is closed, and the second one-way valve (6) is opened, and the nucleic acid solution in the liquid containing chamber (1) flows out from the second one-way valve (6), and flows into the isothermal reaction chamber (7) through the liquid outflow channel (5); S3, when the liquid containing chamber (1) of another microfluidic pump is in an expanded state, the first one-way valve (4) of the microfluidic pump is opened, and the second one-way valve (6) is closed, and the enzyme solution flows in from the liquid inflow channel (3) of the microfluidic pump, and flows into the liquid containing chamber (1) of the microfluidic pump through the first one-way valve (4); when the liquid containing chamber (1) of the microfluidic pump is in a contracted state, the first one-way valve (4) is closed, and the second one-way valve (6) is opened, and the enzyme solution in the liquid containing chamber (1) flows out from the second one-way valve (6), and flows into the isothermal reaction chamber (7) through the liquid outflow channel (5); S4. The nucleic acid solution and the enzyme solution are contacted in the isothermal reaction chamber (7) to perform a nucleic acid isothermal amplification reaction.

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