CN113174323A - Microfluidic PCR chip and PCR detection method - Google Patents

Abstract

The invention discloses a microfluidic PCR chip and a PCR detection method. The reaction is carried out in a closed state to avoid exchange with outside air. The chip includes: a body with a plurality of functional cavities; a rotating chamber, which is rotatably arranged in the body, a transfer chamber is formed in the rotating chamber, and a first communication port is opened on the rotating chamber to communicate with the transfer chamber; and a piston, The body is provided with a closed piston cavity, the piston is movably arranged in the piston cavity, and the piston cavity is divided into a first cavity and a second cavity which are isolated from each other by the piston; the body is also provided with a plurality of liquids respectively communicating with a plurality of functional cavities. a flow channel, a first air flow channel connecting the first cavity and the transfer cavity, and a second air flow channel connecting the second cavity with the inner space of the body except the piston cavity; one end of each liquid flow channel extends to the rotating bin respectively , when the rotating bin is in any working position, one of the liquid flow channels is aligned with the first communication port so that the corresponding functional cavity is communicated with the transfer cavity.

Description

A kind of microfluidic PCR chip and PCR detection method

technical field

The invention belongs to the technical field of biological detection, and relates to a microfluidic PCR chip and a PCR detection method.

Background technique

The microfluidic chip transplants the basic operating units such as sample preparation, mixing, reaction, separation, detection, and cell culture, sorting, and lysis involved in the fields of chemistry or biology onto a small chip, and constructs the whole process. Microchannel network throughout the chip. Microfluidic chips can be used as biological, chemical microreactors or microsystems. Nucleic acid detection has the characteristics of high sensitivity and good specificity, and occupies an extremely important position in life science and medical testing. Combining microfluidic chip technology with nucleic acid detection technology to develop a microfluidic PCR chip that can be used for high-throughput, fully automatic nucleic acid analysis and detection of biological samples, achieving low reagent consumption, fast analysis speed, large detection throughput, With the advantages of high degree of automation and non-professional use, it is especially suitable for multi-index rapid quantitative detection of nucleic acid molecules in biological samples. However, the functional cavity of the existing PCR chip needs to be exchanged with the outside air, and the exhaust gas is discharged directly or after being filtered out of the main body of the chip, which has the risk of polluting the environment.

SUMMARY OF THE INVENTION

The purpose of the present invention is to provide a microfluidic PCR chip, which can integrate nucleic acid extraction, amplification and detection, and conduct the reaction in a closed state, avoid exchange with outside air, and be safe to use.

To achieve the above object, the present invention adopts the following technical solutions:

A microfluidic PCR chip, comprising a body provided with a plurality of functional cavities, the microfluidic chip further comprising:

a rotating chamber, which is rotatably arranged in the body, a transfer cavity is formed in the rotating chamber, and a first communication port communicating with the transfer chamber is opened on the rotating chamber; and

The piston is used to drive the liquid to flow between the functional cavity and the transfer cavity, a closed piston cavity is opened on the body, the piston is movably arranged in the piston cavity, and the piston cavity is the piston is divided into a first cavity and a second cavity isolated from each other;

Wherein, the body is also provided with a plurality of liquid flow channels respectively communicating with the plurality of functional cavities, a first air flow channel connecting the first cavity and the transfer cavity, and a connection between the second cavity and the transfer cavity. a second air flow channel communicating with the inner space of the body except the piston cavity;

One end of each of the liquid flow channels respectively extends to the rotating chamber, and the rotating chamber has a plurality of working positions. In any of the working positions, the liquid flow channel of one of the functional chambers and the The first communication ports are aligned so that the corresponding functional cavity communicates with the transfer cavity.

The inner space refers to a space in the body that is not communicated with the outside world. More preferably, the inner space includes the functional cavity, a microchannel communicating with the functional cavity, and a gap between the functional cavity. The inner space does not include the piston cavity and the cavity communicating with the outer space.

In some preferred embodiments, an annular communication groove is further formed on the body, a second communication port is formed on the rotating bin to communicate with the communication groove, and the shape of the communication groove is the same as that of the first communication groove. The rotation paths of the two communication ports are the same, so that in any of the working positions, the communication groove communicates with the transfer cavity, and the first airflow channel communicates with the communication groove.

In some preferred embodiments, the first communication port is located outside the communication groove.

In some preferred embodiments, the functional chamber includes a reaction chamber, and the second gas flow channel communicates with the reaction chamber.

In some preferred embodiments, there are multiple reaction chambers, and the microfluidic chip includes a control valve capable of connecting or blocking the reaction chamber and the transfer chamber.

In some preferred embodiments, the body is further provided with a plurality of third gas flow channels, each of the reaction chambers is communicated with one of the liquid flow channels and one of the third gas flow channels, respectively, and the control valve A plurality of first micro-channels and a plurality of second micro-channels corresponding to a plurality of reaction chambers are opened on the upper part, the transfer chamber and the piston chamber are located on one side of the control valve, and the reaction chamber is located at the On the other side of the control valve, the control valve is movably inserted in the body to have a communication position and a blocking position. In the communication position, the liquid flow channel of the reaction chamber and the corresponding first A microchannel is connected and communicated, and its third gas flow channel is connected to the second gas flow channel by the corresponding second microchannel; in the blocking position, the liquid flow channel of the reaction chamber is connected to the first gas flow channel. The microchannels are offset from each other, and the third airflow channel and the second microchannel are offset from each other.

In some preferred embodiments, a plurality of the first microchannels and a plurality of the second microchannels are arranged in parallel.

In some preferred embodiments, the first communication port has a circular rotation path, and one end of each of the liquid flow channels extends to the rotation path and is spaced around the rotation path.

In some preferred embodiments, the functional chamber includes a purification chamber, a magnetic rotor is arranged in the purification chamber, and the microfluidic chip further includes a magnet device that drives the magnetic rotor to move.

In some preferred embodiments, the body includes a base layer and a cover layer, the functional cavity is opened on the base layer, the functional cavity includes a sample chamber, and the cover layer covers the base layer The cover plate layer is provided with a sample injection hole communicating with the sample cavity.

The present invention also adopts following technical scheme:

A PCR detection method adopts the above-mentioned microfluidic PCR chip, and the PCR detection method comprises the following steps:

A. Rotate the rotating chamber to make the first communication port communicate with the first functional cavity, and at this time the second airflow channel communicates with the second functional cavity, move the piston to make the liquid flow from the first functional cavity Flow into the transfer cavity of the rotating bin or flow into the first functional cavity from the transfer cavity.

In some preferred embodiments, the PCR detection method further comprises the steps of:

B. Move the control valve so that the first communication port is communicated with the reaction chamber in the functional chamber through the corresponding liquid flow channel. At this time, the second air flow channel is communicated with the reaction chamber, and the piston is moved, making the liquid flow into the reaction chamber from the transfer chamber;

C. Move the control valve to disconnect the liquid flow channel of the reaction chamber, and disconnect the gas flow channel from the reaction chamber.

The present invention adopts the above scheme, has the following advantages compared with the prior art:

The microfluidic PCR chip and the PCR detection method of the present invention selectively connect the functional chamber and the transfer chamber through the rotation of the rotating chamber, and provide positive or negative pressure to the transfer chamber by moving the piston, thereby realizing pipetting; and the piston The two chambers on both sides are connected to the internal chamber of the body, and gas circulation is realized in the chip body, and nucleic acid extraction and nucleic acid amplification are carried out in a fully sealed state to avoid environmental pollution and toxic and harmful substances entering the air through waste gas. It integrates a series of functions such as nucleic acid extraction, purification, amplification and detection, and is easy to operate and safe to use.

Description of drawings

In order to illustrate the technical solutions of the present invention more clearly, the following briefly introduces the accompanying drawings used in the description of the embodiments. Obviously, the accompanying drawings in the following description are only some embodiments of the present invention, which are of great significance to the art For those of ordinary skill, other drawings can also be obtained from these drawings without any creative effort.

FIG. 1 is a three-dimensional schematic diagram of a microfluidic PCR chip according to an embodiment of the present invention from a viewing angle.

FIG. 2 is a schematic perspective view of the microfluidic PCR chip shown in FIG. 1 from another perspective.

FIG. 3 is a top view of the microfluidic PCR chip shown in FIG. 1 .

FIG. 4 is a front view of the microfluidic PCR chip shown in FIG. 1 .

FIG. 5 is a top view of the base layer and the rotating chamber of the microfluidic PCR chip shown in FIG. 1 .

FIG. 6 is a bottom view of the microfluidic PCR chip shown in FIG. 1 .

FIG. 7 is a schematic diagram of a microchannel in the microfluidic PCR chip shown in FIG. 1 .

FIG. 8 is a schematic diagram of the base layer of the microfluidic PCR chip shown in FIG. 1 .

FIG. 9 is a perspective view of a control valve of the microfluidic PCR chip shown in FIG. 1 .

in,

1. Main body; 11. Base layer; 110, Step hole; 111, Step surface; 112, Pipetting hole; 113, Connecting groove; 12, Cover layer; 121, Injection hole;

20, sample chamber; 21, reagent chamber; 22, purification chamber; 23, waste liquid chamber; 24, reaction chamber; 251, first chamber; 252, second chamber; 26, liquid flow channel; 27, first gas flow channel ; 28, the second airflow channel; 29, the third airflow channel;

3. Rotary bin; 30, transfer cavity; 31, first communication port; 32, second communication port; 33, joint groove;

4. Control valve; 41. The first microchannel; 42. The second microchannel;

5. Magnet device.

Detailed ways

The preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings, so that the advantages and features of the present invention can be more easily understood by those skilled in the art. It should be noted here that the descriptions of these embodiments are used to help the understanding of the present invention, but do not constitute a limitation of the present invention.

As shown in this specification and claims, the terms "comprising" and "comprising" only imply that the clearly identified steps and elements are included, and these steps and elements do not constitute an exclusive list. The method or apparatus may also include other steps or elements. As used herein, the term "and/or" includes any combination of one or more of the associated listed items.

It should be noted that, unless otherwise specified, when a feature is called "fixed" or "connected" to another feature, it can be directly fixed or connected to another feature, or it can be indirectly fixed or connected to another feature. on a feature. In addition, the descriptions of upper, lower, left, right, etc. used in the present invention are only relative to the mutual positional relationship of each component of the present invention in the accompanying drawings.

It should be further understood that in the present disclosure, "plurality" refers to two or more, and other quantifiers are similar.

This embodiment provides a microfluidic PCR chip, which integrates nucleic acid extraction and purification, amplification and detection. Referring to FIGS. 1 to 9 , the microfluidic PCR chip includes a body 1 with a plurality of functional cavities, a rotating chamber 3 rotatably arranged in the body 1 , and a piston 13 for driving liquid flow . The functional chamber refers to a chamber in the body 1 for placing the reagents and materials required for the PCR reaction, providing the reaction area required for the reaction, and the like. A transfer chamber 30 is formed in the rotating chamber 3 , and a first communication port 31 communicating with the transfer chamber 30 is opened on the rotating chamber 3 . By rotating the rotating chamber 3, the transfer chamber 30 can be communicated with any functional chamber through a liquid flow channel, thereby realizing liquid transfer.

The body 1 is provided with a closed piston cavity, the piston 13 is movably arranged in the piston cavity, and can perform a reciprocating linear motion in the piston cavity. The piston cavity is separated by the piston 13 and isolated from each other. the first cavity 251 and the second cavity 252; as the piston 13 moves, the first cavity 251 decreases and the second cavity 252 increases, or the first cavity 251 increases and the second cavity 252 decreases, by moving the piston 13 , which can provide positive pressure or negative pressure to the transfer cavity 30, that is, provide the power for liquid circulation.

The body 1 is also provided with a plurality of liquid flow channels 26 respectively communicating with the plurality of functional cavities, a first air flow channel 27 connecting the first cavity 251 with the transfer cavity 30, and a first air flow channel 27 connecting the first cavity 251 with the The second cavity 252 communicates with the second airflow channel 28 of the inner space of the body 1 except the piston cavity. One end of each of the liquid flow channels 26 respectively extends to the rotating chamber 3, and the rotating chamber 3 has a plurality of working positions. In any of the working positions, one of the liquid flow channels 26 and the first A communication port 31 is aligned so that the corresponding functional cavity communicates with the transfer cavity 30 , and the second airflow channel 28 communicates with the inner space in the main body 1 that is not communicated with the outside world. The main body 1 is also provided with an annular communication groove 113, and the rotating bin 3 is provided with a second communication port 32 that communicates with the communication groove 113. The shape of the communication groove 113 is the same as that of the second communication port 32. The rotation paths of the two are the same, so that in any of the working positions, the communication groove 113 communicates with the transfer cavity 30 , and the first airflow channel 27 communicates with the communication groove 113 . That is to say, no matter the rotating bin 3 rotates to any position, the first airflow channel 27 always communicates the first cavity 251 of the piston cavity with the transfer cavity 30 through the communication groove 113 and the second communication port 32 . When the piston 13 moves, the air pressure can be balanced with each other through the gaps in the main body 1 among the above-mentioned functional cavities on the main body 1 . The above-mentioned inner space includes the above-mentioned functional cavity, the microchannel communicating with the functional cavity and the gap between the functional cavities, but does not include the piston cavity and the cavity communicating with the external space.

The chip body 1 is in the shape of a flat plate placed horizontally as a whole; the rotating bin 3 is rotatably embedded in the body 1, and its rotation axis line extends vertically; One end preferably extends out of the body 1 , so as to facilitate the external driving device for driving the movement of the piston 13 . The number of the first communication ports 31 is one, and rotates along a circular rotation path; the number of the second communication ports 32 can be one, two or more, and the distance from the axis of rotation is equal. The first communication port 31 and the second communication port 32 are both opened on the bottom wall of the rotating bin 3, and the second communication port 32 is located inside the first communication port 31; the rotation path of the second communication port 32 is also circular And the rotation paths of the first communication ports 31 form concentric rings with different radii. The communication groove 113 is located below the second communication port 32 . The first airflow channel 27 is opened in the body 1 , one end of which is communicated with the communication groove 113 , and the other end is communicated with the first cavity 251 of the piston 13 . One end of each of the liquid flow channels 26 respectively extends to the rotation path of the first communication port 31 and is arranged at intervals around the cover rotation path.

The functional chamber includes a sample chamber 20, a plurality of reagent chambers 21, a purification chamber 22, a waste liquid chamber 23 and a plurality of reaction chambers 24; the plurality of reaction chambers 24 are arranged side by side on one side of the rotating chamber 3, and the other functional chambers are evenly surrounded Arranged on the other side of the rotating bin 3. The microfluidic PCR chip includes a control valve 4 capable of connecting or blocking the reaction chamber 24 and the transfer chamber 30 . Specifically in this embodiment, the body 1 is further provided with a plurality of third gas flow channels 29 , and each of the reaction chambers 24 is communicated with one of the liquid flow channels 26 and one of the third gas flow channels 29 respectively. The control valve 4 is provided with a plurality of first microchannels 41 and a plurality of second microchannels 42 corresponding to the plurality of reaction chambers 24 respectively. The first microchannels 41 can block the liquid flow channel 26 of the reaction chamber 24 . When it is aligned with a certain liquid flow channel 26, the liquid flow channel 26 is connected, and when it is misaligned with the liquid flow channel 26, the liquid flow channel 26 is blocked by the control valve 4. The transfer chamber 30 and the piston chamber are located on one side of the control valve 4 , and the reaction chamber 24 is located on the other side of the control valve 4 . The control valve 4 is movably inserted in the body 1 to have a communication position and a blocking position. In the communication position, the liquid flow channel 26 of the reaction chamber is connected to the corresponding first microchannel 41 The third airflow channel 29 is connected to the second airflow channel 28 by the corresponding second microchannel 42, and the second airflow channel 28 communicates with each second microchannel 42; in the blocking position , the liquid flow channel 26 and the first microchannel 41 of each of the reaction chambers deviate from each other, and the third airflow channel 29 and the second microchannel 42 deviate from each other. Switching between the communicating position and the blocking position can be achieved by moving the control valve 4 a distance.

The control valve 4 is penetrated in the main body 1 so as to be movable in the horizontal direction, and a plurality of reaction chambers 24 are arranged side by side on the same side of the control valve 4 in sequence along the moving direction of the control valve 4 . A plurality of the first microchannels 41 and a plurality of the second microchannels 42 are arranged in parallel. Specifically, the first microchannel 41 and the second microchannel 42 are parallel to each other, such as extending in the up-down direction. The upper surface of the control valve 4 is provided with a plurality of first holes, and the lower surface of the control valve 4 is provided with a plurality of second holes. The positions and numbers of the first holes and the second holes correspond one by one, and a corresponding group of The first hole and the second hole are respectively used as the inlet and outlet of a first microchannel 41 or a second microchannel 42 .

The body 1 includes a base layer 11 and a cover layer 12 , and the materials of the cover layer 12 and the base layer 11 can be high polymers such as PS, PMMA, PDMA, and PC. The processing methods of the cover layer 12 and the base layer 11 include but are not limited to photolithography, injection molding, machining, laser cutting, etc.; the connection methods of the two include but are not limited to ultrasonic bonding, thermal bonding, anodic bonding, low temperature bonding, etc. The thicknesses of the cover layer 12 and the base layer 11 are respectively 1-20 mm. A stepped hole 110 is provided in the middle of the cover layer 12 , and the hole diameter of the lower part is smaller than that of the upper part. The stepped surface 111 of the stepped hole 110 is respectively provided with a pipetting hole 112 corresponding to each liquid flow channel 26 and the above-mentioned communication groove 113 . The rotating bin 3 is rotatably embedded in the stepped hole 110 . The bottom surface of the rotating bin 3 is provided with an engaging groove 33 for connecting an external driving device, and the external driving device can be a motor for driving the rotating bin 3 to rotate.

Both the functional cavity and the piston cavity are opened on the base layer 11 . The cover layer 12 covers the base layer 11 and is sealed therebetween. The cover layer 12 is provided with a sample injection hole 121 that communicates with the sample cavity 20 ; after sample injection, the sample injection hole 121 can be Seal with lid or film. In addition, there is a space between the cover layer 12 and the base layer 11 that enables communication between the functional cavities, so as to achieve air pressure balance between the functional cavities. The reagent chamber 21 is pre-placed with reagents required for the PCR reaction, such as a lysing solution, a purifying agent, an eluent, etc. Preferably, in this embodiment, the purifying agent is isopropanol. The purification chamber 22 is provided with a magnetic rotor and magnetic beads. The microfluidic PCR chip further includes a magnet device 5 that drives the magnetic rotor to move. The magnet device 5 is close to the purification chamber 22 . The waste liquid chamber 23 is used to recover the waste liquid generated in the process of nucleic acid extraction and the like.

This embodiment also provides a PCR detection method using the microfluidic PCR chip as described above. The PCR detection method comprises the following steps:

A. Rotate the rotating chamber 3 to make the first communication port 31 communicate with the first functional cavity, at this time the second airflow channel 28 communicates with the second functional cavity, move the piston 13, and make the liquid flow from the first functional cavity. One functional cavity flows into the transfer cavity 30 of the rotating bin 3 or flows into the first functional cavity from the transfer cavity 30 .

The PCR detection method also includes the following steps:

B. Move the control valve 4 to make the first communication port 31 communicate with the reaction chamber 24 in the functional chamber through the corresponding liquid flow channel 26. At this time, the second air flow channel 28 communicates with the reaction chamber 24, and moves The piston 13 makes the liquid flow into the reaction chamber 24 from the transfer chamber 30;

C. Move the control valve 4 to disconnect the liquid flow channel 26 of the reaction chamber 24 , and disconnect the gas flow channel from the reaction chamber 24 .

The principle and implementation process of nucleic acid extraction, purification, amplification and detection of the above-mentioned microfluidic PCR chip are as follows:

1. When the rotating bin 3 rotates on the body 1 to different angles, the first communication port 31 of the rotating bin 3 will be connected with the liquid flow channels 26 of different functional chambers on the body 1 .

2. After the above-mentioned liquid flow channels 26 are connected, the piston 13 is moved to realize the transmission of liquid; the precise quantitative function can be realized by precisely moving the piston 13 .

3. After the reagent is added to the purification chamber 22, the magnet device 5 is rotated, and the magnetic rotor in the purification chamber 22 rotates accordingly, thereby realizing the mixing of the magnetic beads.

4. After the liquid is delivered to the 12 reaction chambers 24, move the control valve 4 to completely isolate the reagents in the reaction chambers 24 from the inner chamber.

5. The chambers and microchannels on the base layer 11 are completely sealed by the cover plate layer 12, and all liquid transmission is realized through the piston 13 to achieve internal circulation to prevent contamination.

6. The amplification reaction of PCR reagents is realized by temperature cycle heating, and then detected by the optical module to realize the extraction, amplification and detection of nucleic acid.

This embodiment is an integrated multi-channel microfluidic PCR chip, which not only integrates a series of functions such as nucleic acid extraction, purification, amplification and detection, but also performs nucleic acid extraction and nucleic acid amplification in a fully sealed state. It avoids pollution of the environment; it also meets the needs of multi-channel detection, and also has the characteristics of simple operation process, low reagent consumption, fast analysis speed, etc. It is especially suitable for high-throughput, automatic nucleic acid analysis and detection of biological samples.

The above-mentioned embodiment is only to illustrate the technical concept and characteristics of the present invention, and is a preferred embodiment, and its purpose is that those who are familiar with the technology can understand the content of the present invention and implement it accordingly, and cannot limit the present invention by this. protected range. All equivalent transformations or modifications made according to the principles of the present invention should be included within the protection scope of the present invention.

Claims (12)

1. The utility model provides a micro-fluidic PCR chip, is including the body of seting up a plurality of function chambeies, its characterized in that, micro-fluidic chip still includes:

the rotating bin is rotatably arranged in the body, a transfer cavity is formed in the rotating bin, and a first communication port communicated with the transfer cavity is formed in the rotating bin; and

the piston is used for driving liquid to flow between the function cavity and the transfer cavity, a closed piston cavity is formed in the body, the piston is movably arranged in the piston cavity, and the piston cavity is divided into a first cavity and a second cavity which are mutually isolated by the piston;

wherein, the body is also provided with a plurality of liquid flow channels respectively communicated with the plurality of functional cavities, a first air flow channel for communicating the first cavity with the transfer cavity and a second air flow channel for communicating the second cavity with the inner space of the body except the piston cavity;

one end of each liquid flow channel extends to the rotary bin, the rotary bin is provided with a plurality of working positions, and in any working position, the liquid flow channel of one functional cavity is aligned with the first communication port so that the corresponding functional cavity is communicated with the transfer cavity.

2. The microfluidic PCR chip of claim 1, wherein the body further comprises an annular communicating groove, the rotary chamber comprises a second communicating opening communicating with the communicating groove, the communicating groove has the same shape as the rotary path of the second communicating opening, so that the communicating groove communicates with the transfer chamber at any one of the working positions, and the first air flow channel communicates with the communicating groove.

3. The microfluidic PCR chip of claim 2, wherein the first communication port is located outside the communication groove.

4. The microfluidic PCR chip of claim 1, wherein the functional chamber comprises a reaction chamber, and the second gas flow channel is in communication with the reaction chamber.

5. The microfluidic PCR chip of claim 4, wherein the reaction chamber is multiple, and the microfluidic chip comprises a control valve capable of connecting or disconnecting the reaction chamber and the transfer chamber.

6. The microfluidic PCR chip as claimed in claim 5, wherein the body further comprises a plurality of third gas flow channels, each of the reaction chambers is respectively connected to one of the liquid flow channels and one of the third gas flow channels, the control valve comprises a plurality of first micro channels and a plurality of second micro channels corresponding to the plurality of reaction chambers, the transfer chamber and the piston chamber are located at one side of the control valve, the reaction chamber is located at the other side of the control valve, the control valve is movably inserted into the body and has a communication position and a blocking position, when the control valve is at the communication position, the liquid flow channels of the reaction chamber are connected to and communicated with the corresponding first micro channels, and the third gas flow channels are connected to the second gas flow channels by the corresponding second micro channels; in the blocking position, the flow channel of the reaction chamber and the first microchannel are offset from each other, and the third flow channel and the second microchannel are offset from each other.

7. The microfluidic PCR chip of claim 6, wherein a plurality of the first microchannels and a plurality of the second microchannels are disposed in parallel.

8. The microfluidic PCR chip of claim 1, wherein the first communication port has a circular rotation path, and one end of each of the liquid flow channels extends to the rotation path and is spaced around the rotation path.

9. The microfluidic PCR chip according to claim 1, wherein the functional chamber comprises a purification chamber, a magnetic rotor is disposed in the purification chamber, and the microfluidic chip further comprises a magnet device for driving the magnetic rotor to move.

10. The microfluidic PCR chip of claim 1, wherein the body comprises a substrate layer and a cover plate layer, the functional cavity is disposed on the substrate layer, the functional cavity comprises a sample chamber, the cover plate layer covers the substrate layer and is sealed therebetween, and the cover plate layer is provided with a sample inlet communicated with the sample chamber.

11. A PCR detection method using the microfluidic PCR chip according to any one of claims 1 to 10, comprising the steps of:

A. and rotating the rotary bin to enable the first communication port to be communicated with the first function cavity, at the moment, enabling the second airflow channel to be communicated with the second function cavity, and moving the piston to enable the liquid to flow into the transfer cavity of the rotary bin from the first function cavity or flow into the first function cavity from the transfer cavity.

12. The PCR detection method according to claim 11, further comprising the steps of:

B. a movable control valve which enables the first communication port to be communicated with the reaction cavity in the function cavity through the corresponding liquid flow channel, the second gas flow channel is communicated with the reaction cavity, and the piston is moved to enable liquid to flow into the reaction cavity from the transfer cavity;

C. and moving the control valve to disconnect the liquid flow channel of the reaction cavity, and disconnecting the gas flow channel from the reaction cavity.

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