A fully automatic and fully enclosed fluorescent quantitative PCR microfluidic diagnostic chip

Abstract

The present invention discloses a fully automatic and fully enclosed fluorescent quantitative PCR microfluidic diagnostic chip, including a microfluidic assembly, a plurality of bins are arranged at the top and the bottom, a plurality of microfluidics are arranged on the surface, and each microfluidic is connected to a bin; a flow channel 1 and a flow channel 2 are arranged on the surface of the microfluidic assembly, and the flow channel 1 is connected to the bin with an injection piston arranged in the top; a switching valve installation position is arranged on the microfluidic assembly, and a flow channel switching valve and a reagent switching valve that can rotate relative to it are arranged thereon; a plurality of dry reagent bins are distributed along the circumference of the reagent switching valve, and the dry reagent bins can connect flow channel 1 and flow channel 2 by rotation; an annular common flow channel is arranged on the surface of the flow channel switching valve, and the annular common flow channel is connected to flow channel 2, and a flow channel selection port extending radially along the flow channel switching valve is arranged on the annular common flow channel, and the flow channel selection port can be connected to any microfluidic channel by rotation. The chip integrates nucleic acid extraction, amplification, and detection, reduces pollution, and reduces the difficulty and cost of nucleic acid detection.

Description

Technical Field

[0001] The invention belongs to the field of in vitro diagnosis, in particular to a fully automatic and fully enclosed fluorescent quantitative PCR microfluidic diagnostic chip. Background Art

[0002] The conventional process of nucleic acid testing is: sample sampling, sample preservation and processing, nucleic acid extraction, reagent configuration, nucleic acid amplification, amplification product analysis, amplification product processing and other stages. There are many factors that affect the results of PCR experiments, and one of the most important factors is the nucleic acid aerosol contamination during the PCR experiment. Even trace amounts of nucleic acid aerosol contamination may lead to false positive test results or even failure of the test experiment, so usually all stages of nucleic acid testing need to be completed in a specially set up PCR laboratory. This laboratory must implement partitioned operations for sample processing, nucleic acid extraction, and PCR amplification, and must have a good ventilation system. The cost of setting up a laboratory is very high, and often only large medical institutions have the financial resources to build one.

[0003] Microfluidics refers to the technology of manipulating trace fluids using tiny channels and various microstructures. It is an emerging interdisciplinary subject involving chemistry, fluid physics, microelectronics, new materials, biology and biomedical engineering. Because of its miniaturization and integration, microfluidics devices are usually called microfluidics chips or chip laboratories. A small microfluidics chip is equivalent to integrating the basic operations of sample preparation, reaction, separation, detection and so on in the biological, chemical and medical analysis process, and automatically completing the whole process of analysis with the help of instruments. Microfluidics chips generally have the advantages of low sample consumption, fast detection speed, simple operation, multi-functional integration, small size and easy to carry. Therefore, they have great potential to simplify the diagnostic process, reduce medical costs and improve medical efficiency.

[0004] If microfluidics technology can be used in the nucleic acid testing process, the requirements for laboratories will be greatly reduced, and the difficulty and cost of nucleic acid testing will be reduced. It will also be conducive to the promotion of nucleic acid testing to grassroots hospitals, and will make a huge contribution in particular to the detection of various infectious diseases. Summary of the invention

[0005] The technical problem solved by the present invention is to provide a fully enclosed fluorescent quantitative PCR microfluidic diagnostic chip integrating nucleic acid extraction, amplification and detection, thereby reducing pollution and lowering the difficulty and cost of nucleic acid detection.

[0006] The technical solution adopted by the present invention is as follows:

[0007] A fully automatic and fully enclosed fluorescent quantitative PCR microfluidic diagnostic chip, comprising a microfluidic assembly, wherein a plurality of bins are arranged at the top and the bottom of the microfluidic assembly, a plurality of microfluidics are arranged on the surface of the microfluidic assembly, and each microfluidic channel is connected to a bin; a flow channel 1 and a flow channel 2 are also arranged in the middle of the surface of the microfluidic assembly, wherein the flow channel 1 is connected to one of the bins at the top, and an injection piston is arranged in the bin;

[0008] The microfluidic component is also provided with a switching valve installation position, and the switching valve installation position is provided with a channel switching valve and a reagent switching valve that can rotate relative to the microfluidic component; a plurality of dry reagent compartments are distributed along the circumference of the reagent switching valve, and the dry reagent compartments can connect channel one and channel two by rotation; an annular common channel is provided on the surface of the channel switching valve, and the annular common channel is connected with channel two, and the annular common channel is provided with a channel selection port extending along the radial direction of the channel switching valve, and the channel selection port can be connected with any microfluidic channel by rotation.

[0009] Preferably, the chamber body at the top of the flow channel assembly includes a liquid transfer chamber, a plurality of reagent chambers, and a sample chamber, and an injection piston is provided in the liquid transfer chamber; the chamber body at the bottom of the flow channel assembly includes a PCR amplification detection chamber, and a reaction chamber, and the reaction chamber includes a nucleic acid extraction chamber and a multifunctional chamber that are interconnected.

[0010] Preferably, a gas internal circulation hole is also provided on the upper portion of the surface of the microfluidic channel component, the gas internal circulation hole is connected to the gas microfluidic channel, and the gas microfluidic channel is connected to the multifunctional chamber.

[0011] Preferably, a ventilation channel is also provided on the surface of the flow channel switching valve, and the microchannel connected to the gas circulation inner hole is a three-way type. The ventilation channel can be connected to the free end of the three-way microchannel by rotating the flow channel switching valve, and the ventilation channel is connected to the PCR amplification detection chamber through the three-way microchannel.

[0012] Preferably, the microfluidic channel assembly further comprises a reagent sealing film, and the reagent sealing film is arranged on the top of the plurality of reagent chambers.

[0013] Preferably, the microfluidic assembly includes a chip body and a channel sealing layer assembled as one, and a plurality of chambers and microchannels, as well as channel one and channel two are arranged on the chip body; the channel sealing layer is provided with a reagent chamber inlet and a reagent chamber outlet, the reagent chamber inlet is connected with channel one, and the reagent chamber outlet is connected with channel two, and the reagent switching valve connects the dry reagent chamber with the reagent chamber inlet and the reagent chamber outlet by rotation.

[0014] Preferably, the chip body and the flow channel sealing layer are combined together by any connection method such as ultrasonic welding, bonding or hot melting to form a complete flow channel to form a microfluidic channel component.

[0015] Preferably, the PCR microfluidic diagnostic chip also includes a chip upper shell assembly arranged on the upper part of the microfluidic assembly, the chip upper shell assembly includes a chip upper shell, a chip sealing cover is arranged on the chip upper shell, and the chip sealing cover corresponds to the sample chamber; a sealing ring is arranged between the chip upper shell assembly and the microfluidic assembly.

[0016] Preferably, the chip upper shell assembly further comprises a sealing elastic film and a sealing film pressing plate, the sealing film pressing plate is provided with a plurality of through holes, the through holes correspond to the reagent chambers, and the sealing film pressing plate is further provided with a sealing elastic film.

[0017] Preferably, the PCR microfluidic diagnostic chip further comprises a chip lower shell matching the microfluidic channel assembly.

[0018] The beneficial technical effects of the present invention are:

[0019] By applying microfluidic technology to nucleic acid detection chips, the chip has shrunk the PCR laboratory that costs huge amounts of money to build for traditional PCR experiments onto a small test chip, and has automated the detection, greatly reducing the requirements for the laboratory. It also reduces the difficulty and cost of nucleic acid testing, and is conducive to the promotion of nucleic acid testing to grassroots hospitals, making a huge contribution in particular to the detection of various infectious diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 Schematic diagram of the chip appearance;

[0021] Figure 2 It is a schematic diagram of chip components;

[0022] Figure 3 It is a schematic diagram of the chip shell assembly;

[0023] Figure 4 Schematic diagram of the chip microfluidic assembly;

[0024] Figure 5 Schematic diagram of the main structure of the chip.

[0025] The names corresponding to the reference numerals are:

[0026] 1-chip upper shell assembly, 2-microfluidic assembly, 3-chip lower shell three, 1-1-chip sealing cover, 1-2-chip upper shell, 1-3-sealing elastic film, 1-4-sealing film pressing plate, 2-1-chip body, 2-2-sealing ring, 2-3-reagent sealing film, 2-4-injection piston, 2-5-channel switching valve, 2-6-channel sealing layer, 2-7-reagent switching valve, 2-8-lyophilized reagent, 2-1-1-liquid transfer chamber, 2-1-2-reagent chamber, 2-1-3-sample chamber, 2-1-4-gas internal circulation hole, 2-1-5-microchannel, 2-1-6-switching valve installation position, 2-1-7-PCR amplification detection chamber, 2-1-8-nucleic acid extraction chamber, 2-1-9-multi-function chamber, 2-1-10-channel one, 2-1-11 channel two, 2-5-1 annular common flow channel, 2-5-2 channel selection port, 2-5-3 ventilation channel, 2-6-1 reagent chamber inlet, 2-6-2 reagent chamber outlet. DETAILED DESCRIPTION

[0027] The present invention is further described below in conjunction with the accompanying drawings and embodiments. The embodiments of the present invention include but are not limited to the following embodiments.

[0028] The present invention uses microfluidic technology to integrate the basic operations of sample preparation, reaction, separation, detection, etc. in biological, chemical, and medical analysis processes, and automatically completes the entire analysis process with the instrument. The specific structure is as follows Figure 1-5 As shown,

[0029] A fully automatic and fully enclosed fluorescent quantitative PCR microfluidic diagnostic chip, comprising a microfluidic component 2, wherein a plurality of bins are arranged at the top and the bottom of the microfluidic component 2, a plurality of microfluidic channels 2-1-5 are arranged on the surface of the microfluidic component 2, and each microfluidic channel 2-1-5 is connected to a bin; a flow channel 1 2-1-10 and a flow channel 2 2-1-11 are also arranged in the middle of the surface of the microfluidic component 2, the flow channel 1 2-1-10 is connected to one of the bins at the top, and an injection piston 2-4 is arranged in the bin;

[0030] The microfluidic component 2 is also provided with a switching valve installation position 2-1-6, and the switching valve installation position 2-1-6 is provided with a channel switching valve 2-5 and a reagent switching valve 2-7 that can rotate relative to the microfluidic component 2; the reagent switching valve 2-7 is provided with a plurality of dry reagent compartments distributed along the circumference, and the dry reagent compartments can connect channel one 2-1-10 and channel two 2-1-11 by rotation; the surface of the channel switching valve 2-5 is provided with an annular common channel 2-5-1, and the annular common channel 2-5-1 is connected with channel two 2-1-11, and the annular common channel 2-5-1 is provided with a channel selection port 2-5-2 extending along the radial direction of the channel switching valve, and the channel selection port 2-5-2 can be connected with the microfluidic channel 2-1-5 by rotation.

[0031] Furthermore, when conducting nucleic acid detection, the chamber body at the top of the flow channel component is divided into a liquid transfer chamber 2-1-1, a plurality of reagent chambers 2-1-2, and a sample chamber 2-1-3, and an injection piston 2-4 is arranged in the liquid transfer chamber 2-1-1; the chamber body at the lower part of the flow channel component 2 includes a PCR amplification detection chamber 2-1-7, and a reaction chamber, and the reaction chamber includes a nucleic acid extraction chamber 2-1-8 and a multi-functional chamber 2-1-9 which are interconnected.

[0032] In the above structure, the liquid transfer chamber 2-1-1 is used for mixing and transferring various liquid reagents and freeze-dried reagent balls; the reagent chamber 2-1-2 is used for pre-packaging of various liquid or solid reagents or as a reserved empty chamber; the sample chamber 2-1-3 is used for users to add test samples; the microchannel 2-1-5 is used for liquid flow; the switching valve installation position 2-1-6 is used to install the channel switching valve 2-5 and the reagent switching valve 2-7; the PCR amplification detection chamber 2-1-7 is used for amplification reaction and fluorescence signal detection; the nucleic acid extraction chamber 2-1-8 is used to separate and extract nucleic acids; the multi-functional chamber 2-1-9 is used for liquid heating, liquid mixing, nucleic acid lysis, nucleic acid elution, etc.

[0033] The microfluidic assembly of the above structure is assembled with an injection piston 2-4, a reagent switching valve 2-7, a channel switching valve 2-5 and a freeze-dried reagent 2-8; wherein the freeze-dried reagent 2-8 is placed in the dry reagent compartment of the reagent switching valve 2-7, and the reagent switching valve 2-7 and the channel switching valve 2-5 are rotatably connected to the switching valve mounting position 2-1-6 on the microfluidic assembly, and the dry reagent compartment can be connected to the channel 1 2-1-10 and the channel 2 2-1-11 on the surface of the microfluidic assembly by rotating the reagent switching valve 2-7; and the channel switching valve 2-5 can be rotated to enable the channel selection port 2-5-2 to be connected to the selected microfluidic channel 2-1-5.

[0034] The above-mentioned PCR microfluidic diagnostic chip can be operated in the following manner: after the sample is injected into the bottom of the sample chamber 2-1-3, it can be injected slowly to avoid the generation of bubbles, and then the piston rod is inserted into the bottom of the liquid transfer chamber 2-1-1 and the injection piston 2-4 is assembled, and then the rotating channel switching valve 2-5 and the reagent switching valve 2-7 are driven respectively, and the channel selection port 2-5-2 on the rotating channel switching valve 2-5 is connected to the sample chamber 2-1-3, and the dry reagent chamber containing the freeze-dried magnetic bead reagent in the rotating reagent switching valve 2-7 is connected to the channel one 2-1-10 and the channel two 2-1-11, so that the liquid transfer chamber 2-1-1, the channel one 2-1-10, the dry reagent chamber, the channel two 2-1-11, the annular common channel 2-5-1, the channel selection port 2-5-2, and the sample chamber 2-1-3 are connected, and the piston rod and The injection piston 2-4 cooperates to absorb a quantitative sample and mix it with the freeze-dried magnetic bead reagent and reach the liquid transfer chamber 2-1-1; then the channel switching valve 2-5 is driven again to connect the microchannel 2-1-5 corresponding to the reagent chamber 2-1-2 containing the lysis solution, that is, the liquid transfer chamber 2-1-1 is connected with the reagent chamber 2-1-2 containing the lysis solution, and then a quantitative lysis solution is absorbed into the liquid transfer chamber 2-1-1 through the injection piston and preliminarily mixed with the sample in the chamber; then the channel switching valve 2-5 is driven again to connect the nucleic acid extraction chamber 2-1-8 and the multi-functional chamber 2-1-8, and the injection piston 2-4 is driven to discharge the mixed liquid from the liquid transfer chamber 2-1-1 into the multi-functional chamber 2-1-9, and at the same time, the outside of the multi-functional chamber 2-1-9 is heated at a constant temperature. During the heating process, the injection piston 2-4 can repeatedly draw liquid to accelerate the lysis of cells.After the lysis is completed, the nucleic acid in the liquid is combined with the magnetic beads. By placing a permanent magnet outside the nucleic acid extraction chamber 2-1-8 or using an electromagnet to generate strong magnetism, when the injection piston 2-4 slowly extracts the liquid from the inside of the multi-functional chamber 2-1-9 and transfers it to the liquid transfer chamber 2-1-1, the magnetic beads and the combination of magnetic beads and nucleic acid in the liquid are adsorbed in the nucleic acid extraction chamber 2-1-8; then, the flow channel switching valve 2-5 is driven to connect the sample chamber 2-1-3 and discharge the liquid. After the sample chamber 2-1-3 is full, the flow channel switching valve 2-5 is driven to connect the reagent chamber 2-1-2 with the lysis solution, and the remaining waste liquid is discharged therein; then, the flow channel switching valve 2-5 is continued to be driven to connect the reagent chamber 2-1-2 with the cleaning solution, and the liquid transfer chamber 2-1-1 is sucked in, and then the flow channel switching valve 2-5 is driven to connect the liquid transfer chamber 2-1-1 with the nucleic acid extraction chamber 2-1-8, and the magnetic beads in the nucleic acid extraction chamber 2-1-8 are cleaned multiple times with the cleaning solution, and then the magnetic beads are adsorbed. The waste liquid is discharged to the reagent compartment 2-1-2 with a cleaning liquid; then the eluent in the reagent compartment 2-1-2 with an eluent is quantitatively drawn to elute the nucleic acid on the magnetic beads in the nucleic acid extraction compartment 2-1-8, and the magnetic beads are separated to obtain the eluent which is stored in the liquid transfer compartment 2-1-1; then the flow channel switching valve 2-5 is driven to connect to the microchannel 2-1-5 with an empty chamber reserved in the reagent compartment 2-1-2, and the reagent switching valve 2-7 is driven to connect to the dry reagent compartment where the PCR reagent is located, so that the eluent and the P After the CR reagent is fully mixed, it is sucked into the liquid transfer chamber 2-1-1, and finally the flow channel switching valve 2-5 is driven to connect to the PCR amplification detection chamber 2-1-7, and the mixed liquid in the liquid transfer chamber 2-1-1 is injected into the PCR amplification detection chamber 2-1-7, and the inlet and outlet channels of the PCR amplification detection chamber 2-1-7 are closed. At the same time, the instrument performs cyclic temperature heating on the PCR amplification detection chamber 2-1-7 through an external heating device, and detects real-time fluorescence signals through an external fluorescence device, and finally completes the detection to obtain the results.

[0035] Furthermore, a gas internal circulation hole 2-1-4 is also provided on the upper surface of the microfluidic assembly 2, and the gas internal circulation hole 2-1-4 is connected to the microfluidic channel, and the microfluidic channel is connected to the multifunctional chamber 2-1-9. In the above structure, the multifunctional chamber can be ventilated according to actual conditions, thereby increasing the application range of the entire PCR microfluidic diagnostic chip.

[0036] Furthermore, the surface of the flow channel switching valve 2-5 is also provided with a ventilation flow channel 2-5-3, and the microchannel connected to the gas circulation inner hole 2-1-4 is a three-way type. The ventilation flow channel 2-5-3 can be connected to the free end of the three-way microchannel by rotating the flow channel switching valve 2-5, and the ventilation flow channel 2-5-3 is connected to the PCR amplification detection chamber 2-1-7 through the three-way microchannel. The above structure can realize the connection between the PCR amplification detection chamber 2-1-7 and the atmosphere. When the flow channel of the PCR amplification detection chamber 2-1-7 is closed, it can be connected to the atmosphere by rotating the flow channel switching valve 2-5. Whether to connect can be selected according to the actual situation. It can be connected to the atmosphere, and other gases can also be connected through the gas circulation inner hole 2-1-4 to increase the application range of the PCR microfluidic diagnostic chip.

[0037] Furthermore, the microfluidic assembly 2 also includes a reagent sealing film 2-3, which is arranged on the top of the plurality of reagent chambers 2-1-2. It is used for sealing to prevent the reagent from being contaminated during assembly. When the reagent in the reagent chamber 2-1-2 is to be used, the reagent sealing film 2-3 corresponding to the column can be punctured.

[0038] Furthermore, the microfluidic assembly 2 includes a chip body 2-1 and a flow channel sealing layer 2-6 assembled as one body, and the chip body 2-1 is provided with a plurality of chambers and microfluidics 2-1-5, as well as flow channel 1 2-1-10 and flow channel 2 2-1-11; the flow channel sealing layer 2-6 is provided with a reagent chamber inlet 2-6-1 and a reagent chamber outlet 2-6-2, the reagent chamber inlet 2-6-1 is connected with flow channel 1 2-1-10, the reagent chamber outlet 2-6-2 is connected with flow channel 2 2-1-11, and the reagent switching valve 2-7 is rotated to connect the dry reagent chamber with the reagent chamber inlet 2-6-1 and the reagent chamber outlet 2-6-2. The above structure describes the specific structure of the microfluidic assembly, and the structure of how flow channel 1 2-1-10 and flow channel 2 2-1-11 are connected with the dry reagent chamber.

[0039] Furthermore, the chip body 2 - 1 and the flow channel sealing layer 2 - 6 are combined together by any connection method such as ultrasonic welding, bonding or hot melting to form a complete flow channel to form a microfluidic channel component.

[0040] Furthermore, the PCR microfluidic diagnostic chip also includes a chip shell assembly 1 arranged on the upper part of the microfluidic assembly 2, the chip shell assembly 1 includes a chip shell 1-2, a chip sealing cover 1-1 is arranged on the chip shell 1-2, and the chip sealing cover 1-1 corresponds to the sample chamber 2-1-3; a sealing ring 2-2 is arranged between the chip shell assembly 1 and the microfluidic assembly 2.

[0041] Furthermore, the chip shell assembly 1 also includes a sealing elastic film 1-3 and a sealing film pressing plate 1-4, the sealing film pressing plate 1-4 is provided with a plurality of through holes, the through holes correspond to the reagent chamber 2-1-2, and the sealing elastic film 1-3 is also provided on the sealing film pressing plate 1-4.

[0042] The present invention, when producing the chip, pre-packages various reagents required for the entire molecular diagnosis process in the reagent chamber 2-1-2 and the reagent switching valve 2-7 provided in the microfluidic channel assembly 2, and interconnects with each functional unit through the microfluidic channel 2-1-5, and provides an injection piston 2-4 for driving the flow of liquid and a flow channel switching valve 2-5 for switching the flow channel. At the same time, the microfluidic channel assembly 2 is also provided with an assembly chip upper shell assembly 1, and a sealing elastic film 1-3, a chip sealing cover 1-1, and a sealing ring 2-2 between the two are provided through the chip upper shell assembly 1, all of which are used to seal the chip, so that the chip becomes a closed space, and the air in the chip forms an internal circulation, thereby avoiding mutual contamination of aerosols inside and outside the chip. When the above-mentioned chip upper shell assembly 1 is provided, when the chip is used, when the reagent sealing film 2-3 is pierced by a piercing column, it is separated by the sealing elastic film 1-3, and the sealing elastic film 1-3 is not pierced, which can play a good sealing role in the detection process.

[0043] Furthermore, the PCR microfluidic diagnostic chip further comprises a chip lower shell 3 matched with the microfluidic channel assembly 2. The chip lower shell 3 is used for assembling and sealing the chip.

[0044] The above is only a preferred specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be easily thought of by a person skilled in the art within the technical scope disclosed in the present invention should be covered within the protection scope of the present invention. The present invention extends to any new features or any new combination disclosed in this specification, as well as any new method or process steps or any new combination disclosed.

Claims

1. A fully automatic and fully enclosed fluorescent quantitative PCR microfluidic diagnostic chip, characterized in that: It comprises a microfluidic channel assembly, wherein a plurality of bins are arranged at the top and the bottom of the microfluidic channel assembly, a plurality of microfluidics are arranged on the surface of the microfluidic channel assembly, and each microfluidic channel is connected to a bin; a flow channel 1 and a flow channel 2 are also arranged in the middle of the surface of the microfluidic channel assembly, and the flow channel 1 is connected to one of the bins at the top, and an injection piston is arranged in the bin; The microfluidic assembly is also provided with a switching valve installation position, and the switching valve installation position is provided with a channel switching valve and a reagent switching valve that can rotate relative to the microfluidic assembly; a plurality of dry reagent compartments are distributed along the circumference of the reagent switching valve, and the dry reagent compartments can connect channel one and channel two by rotation; an annular common channel is provided on the surface of the channel switching valve, and the annular common channel is connected with channel two, and the annular common channel is provided with a channel selection port extending along the radial direction of the channel switching valve, and the channel selection port can be connected with any microfluidic channel by rotation; The chamber body at the top of the flow channel assembly includes a liquid transfer chamber, a plurality of reagent chambers, and a sample chamber, and an injection piston is arranged in the liquid transfer chamber; the chamber body at the bottom of the flow channel assembly includes a PCR amplification detection chamber, and a reaction chamber, and the reaction chamber includes a nucleic acid extraction chamber and a multifunctional chamber that are interconnected; a gas inner circulation hole is also arranged on the upper surface of the microfluidic channel assembly, and the gas inner circulation hole is connected to the gas microfluidic channel, and the gas microfluidic channel is connected to the multifunctional chamber; a ventilation flow channel is also arranged on the surface of the flow channel switching valve, and the microfluidic channel connected to the gas circulation inner hole is a three-way type, and the ventilation flow channel can be connected to the free end of the three-way type microfluidic channel by rotating the flow channel switching valve, and the ventilation flow channel is connected to the PCR amplification detection chamber through the three-way type microfluidic channel; The microfluidic component includes a chip body and a channel sealing layer assembled as one, and a plurality of chambers and microfluidics, as well as channel one and channel two are arranged on the chip body; the channel sealing layer is provided with a reagent chamber inlet and a reagent chamber outlet, the reagent chamber inlet is connected with channel one, and the reagent chamber outlet is connected with channel two, and the reagent switching valve connects the dry reagent chamber with the reagent chamber inlet and the reagent chamber outlet by rotation.

2. A fully automatic and fully enclosed fluorescent quantitative PCR microfluidic diagnostic chip according to claim 1, characterized in that: The microfluidic channel assembly further comprises a reagent sealing film, and the reagent sealing film is arranged on the top of the plurality of reagent compartments.

3. A fully automatic and fully enclosed fluorescent quantitative PCR microfluidic diagnostic chip according to claim 1, characterized in that: The chip body and the flow channel sealing layer are combined together by any connection method such as ultrasonic welding, bonding or hot melting to form a complete flow channel to form a micro-flow channel component.

4. A fully automatic and fully enclosed fluorescent quantitative PCR microfluidic diagnostic chip according to any one of claims 1 to 3, characterized in that: The PCR microfluidic diagnostic chip also includes a chip upper shell component arranged on the upper part of the microfluidic channel component, the chip upper shell component includes a chip upper shell, a chip sealing cover is arranged on the chip upper shell, and the chip sealing cover corresponds to the sample chamber; a sealing ring is arranged between the chip upper shell component and the microfluidic channel component.

5. A fully automatic and fully enclosed fluorescent quantitative PCR microfluidic diagnostic chip according to claim 4, characterized in that: The chip upper shell component also includes a sealing elastic film and a sealing film pressing plate. The sealing film pressing plate is provided with a plurality of through holes corresponding to the reagent chambers. The sealing film pressing plate is also provided with a sealing elastic film.

6. A fully automatic and fully enclosed fluorescent quantitative PCR microfluidic diagnostic chip according to claim 5, characterized in that: The PCR microfluidic diagnostic chip also includes a chip lower shell matching the microfluidic channel assembly.

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