CN216237017U - Closed micro-fluidic chip for PCR reaction - Google Patents

Abstract

The utility model relates to the field of biomedical science, in particular to a closed microfluidic chip for PCR reaction, which comprises a gland and at least one chip, wherein one side of the gland is provided with at least one mounting groove, a buckle, at least one gland sample inlet and at least one gland gas outlet, and the other side of the gland is provided with a sealing film; the chip is installed in the installation groove through the buckle; a sealing ring containing a micro-cavity hole is arranged between the chip and the gland, the micro-cavity hole is a liquid inlet hole and an air outlet hole, the liquid inlet hole is communicated with the sample inlet of the gland, and the air outlet hole is communicated with the air outlet of the gland; the sealing film covers each gland sample inlet and gland gas outlet. The utility model can realize the totally-enclosed PCR amplification environment and avoid pollution; the temperature can be rapidly increased and decreased, and the detection time is shortened; the difficulty and the cost of manufacturing the chip are reduced.

Description

Closed micro-fluidic chip for PCR reaction

Technical Field

The utility model relates to the field of biological medical treatment, in particular to a closed microfluidic chip for PCR reaction.

Background

As one of the main detection modes in the field of in vitro diagnosis, nucleic acid detection is the most direct, reliable and sensitive method for realizing early, rapid and specific detection of pathogens, can rapidly detect pathogen nucleic acid in a detection sample, and provides scientific detection basis for accurate diagnosis of infectious cases. The nucleic acid amplification detection is to amplify a nucleic acid sequence to be detected through the action of enzyme, wherein the PCR (polymerase chain reaction) technology is most widely applied due to the advantages of good specificity, low cost and the like. PCR consists of three basic reaction steps of denaturation, annealing and extension, the reaction time is always the limiting factor of the application of the PCR technology, and an instrument improved aiming at the PCR reaction time is presented at present, for example, patent document CN 111269825A discloses a rapid nucleic acid instant detector with a rapid temperature raising and lowering module, but the current PCR reaction container has lower heat conduction efficiency, and for the PCR reaction needing circulating temperature raising and lowering, the PCR reaction container has lower heat conduction efficiency and slow temperature raising and lowering speed, so the whole process takes longer time.

The micro-fluidic chip technology integrates the traditional biochemical analysis on a chip with the size of a few square centimeters or even smaller, and completes detection and analysis in a micro-nano scale channel and a micro-chamber in the chip. However, the microfluidic chip requires a suitable heat-conducting material in nucleic acid detection, so that the manufacturing difficulty and cost are greatly increased, and the totally-enclosed PCR amplification environment is difficult to realize, thereby limiting the further clinical application of the microfluidic chip. Therefore, a closed microfluidic chip with high thermal conductivity is urgently needed, the thermal conductivity of the microfluidic chip is improved, the detection time is shortened, the totally closed PCR amplification environment can be realized, and the manufacturing cost of the chip is reduced.

SUMMERY OF THE UTILITY MODEL

In view of the whole or partial defects of the prior art, the utility model provides the closed microfluidic chip for PCR reaction, which realizes a totally closed PCR amplification environment and avoids pollution by arranging the sealing ring, the iron sheet and the buckle structure; meanwhile, the heating and cooling of the amplification cavity can be synchronous with the heating and cooling of an external heat source by arranging the heat-conducting fins, so that the heating and cooling are realized quickly, and the time consumption of nucleic acid detection is greatly shortened; in addition, the utility model adopts the combination of the 8-inch wafer etching process and the injection molding production of the gland, has low production cost and is suitable for large-scale application and popularization.

In order to achieve the purpose of the utility model, the utility model provides the following technical scheme: a closed microfluidic chip for PCR reaction is characterized in that: the device comprises a gland and at least one chip, wherein one side of the gland is provided with at least one mounting groove, a buckle, at least one gland sample inlet and at least one gland gas outlet, and the other side of the gland is provided with a sealing film; the chip is installed in the installation groove through the buckle; a sealing ring containing a micro-cavity hole is arranged between the chip and the gland, the micro-cavity hole is a liquid inlet hole and an air outlet hole, the liquid inlet hole is communicated with a gland sample inlet, and the air outlet hole is communicated with a gland air outlet; the sealing film covers each gland sample inlet and gland gas outlet. An iron sheet is embedded in the press cover through an injection molding process, and the iron sheet interacts with a magnet on external equipment to generate attraction. The technical scheme has the beneficial effects that through the arrangement of the sealing ring with the micro-cavity hole, on one hand, the elastic buffering between the chip and the gland can be realized, so that the chip can effectively contact the external heat transfer seat without generating an air cavity; on the other hand, the arranged iron sheet and the external equipment are provided with magnets which interact to generate attraction, the buckle can generate pressure when fixing the chip, the attraction and the pressure simultaneously act to enable the sealing ring to receive double acting force in the vertical direction, the sealing effect is further enhanced, and the reaction liquid cannot leak into the air to cause aerosol pollution in the temperature rising and falling process; meanwhile, the sealing film covers all the gland sample inlets and all the gland gas outlets, so that a totally-enclosed PCR amplification environment is realized.

The sealing film is made of biocompatible and malleable materials, and the materials can resist temperature for more than 0.5h at the temperature of 100 ℃. The technical scheme has the beneficial effects that the adopted material with biocompatibility and ductility can not damage the sample during the PCR reaction; on the other hand, the material has the temperature resistance time of more than 0.5h at the temperature of 100 ℃, has good high temperature resistance property, and the sealing performance of the material cannot be weakened by heat released in the PCR reaction process, so that the sealing in the whole PCR reaction process is ensured.

The chip is provided with a first structure body, a second structure body and a heat conducting fin from top to bottom, wherein the first structure body and the second structure body are both of sheet structures; the heat conducting sheet is made of heat conducting materials with heat conducting coefficients not lower than 50W/(m.K), and the thickness of the heat conducting sheet is 0.05mm-0.2 mm. The technical scheme has the beneficial effects that the heat is transferred to the heat conducting sheet through the external heat transfer seat, and the heat conducting sheet transfers the heat to the second structure body of the chip, so that the temperature rise and fall of the amplification cavity in the chip and the temperature rise and fall of the heat transfer seat are almost same in frequency, the rapid temperature rise and fall of the micro-fluidic chip are realized, and the time consumption of nucleic acid detection is greatly shortened. The heat conduction coefficient is not lower than 50W/(m.K), the heat conduction material is used as the material of the heat conduction sheet, the heat transfer efficiency of the whole chip can be further improved, the thickness of the heat conduction sheet can also influence the heat transfer efficiency, and the thickness of the heat conduction sheet is 0.05mm-0.2mm, so that the heat conduction sheet can play a role of heat transfer and buffering to the maximum extent.

The second structure body is provided with at least one amplification cavity, at least one liquid inlet micro-channel and at least one liquid outlet micro-channel. The technical scheme has the beneficial effects that the second structure body can be provided with a plurality of amplification cavities, so that the detection of one to a plurality of sample nucleic acids can be realized simultaneously, and the nucleic acid detection efficiency is improved.

An elastic part is further arranged between the chip and the gland, the elastic part is arranged on the other side of the sealing ring with the micro-cavity hole, and the elastic part and the sealing ring with the micro-cavity hole enable the chip to be horizontally arranged on the gland. This technical scheme beneficial effect lies in, forms balanced state through setting up the elastic component and the sealing washer that contains the microcavity hole for the more effective outside heat transfer seat of contact of chip does not produce the air cavity.

The gland is provided with four mounting grooves, a chip is assembled in each mounting groove, and each chip is provided with one amplification cavity; the first structure body of the chip is a glass sheet, and the second structure body of the chip is a silicon sheet; the two sides of the gland are symmetrically provided with handheld structures, and the two iron sheets are symmetrically placed on the two sides of the gland; the gland is provided with a positioning hole, and the gland is positioned on external equipment through the positioning hole. The technical scheme has the beneficial effects that the heat transfer seat can be accurately positioned at the heat source of an external instrument through the arrangement of the positioning hole, so that the temperature rising and reducing efficiency is improved; meanwhile, four chips are assembled on one gland, and a reaction cavity is arranged on each chip, so that samples 1-4 can be detected simultaneously, and the nucleic acid detection efficiency is improved.

The amplification cavity is a coiled pipe amplification cavity. The technical scheme has the beneficial effects that the amplification cavity and the amplification cavity are arranged in a snake shape, so that the contact area between the sample and the chip can be increased, and the realization of temperature rise and fall at the same frequency is facilitated.

The technical scheme of the utility model has the following beneficial effects: according to the multi-material combined microfluidic chip for PCR rapid reaction, the sealing effect is enhanced through the combination of the buckle structure, the ferromagnetic metal and the sealing ring; the heat conducting sheet made of the heat conducting material is fixed with the heat conducting seat through the buckle structure, so that the reaction cavity is quickly heated and cooled, and the time consumed by nucleic acid detection is greatly shortened; meanwhile, the 8-inch wafer etching process is combined with injection molding production of the gland, so that the manufacturing process is simple, the cost is low, and large-scale application and popularization are facilitated.

Drawings

In order to more clearly illustrate the technical solutions in the specific embodiments of the present invention, the drawings needed to be used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings described below are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings can be obtained according to these drawings without inventive efforts.

Fig. 1 is a schematic bottom perspective view of a closed microfluidic chip according to a first embodiment of the present invention;

fig. 2 is a schematic bottom view of a closed microfluidic chip and an external heat transfer seat according to a first embodiment of the present invention;

fig. 3 is a schematic front view of a closed microfluidic chip according to a first embodiment of the present invention;

fig. 4 is a left side view schematically illustrating a closed microfluidic chip according to a first embodiment of the present invention;

fig. 5 is a schematic top perspective view of a gland of the closed microfluidic chip of fig. 1; fig. 6 is a schematic bottom perspective view of the gland and the external heat transfer seat of the enclosed microfluidic chip of fig. 1;

fig. 7 is a schematic diagram of an internal structure of a chip of the closed microfluidic chip of fig. 1;

fig. 8 is a bottom perspective view of the chip of the enclosed microfluidic chip of fig. 1;

fig. 9 is a schematic front view of a chip of the closed microfluidic chip of fig. 1.

Reference numerals: 1-a gland, 2-an iron sheet, 3-a positioning hole, 4-a sealing film, 5-a chip, 6-a sealing ring containing a micro-cavity hole, 7-a buffer silica gel gasket, 8-a heat transfer seat, 9-a mounting groove, 101, 102-a mounting groove left side buckle, 103, 104-a mounting groove right side buckle, 105-a mounting groove hole side buckle, 106-a gland sample inlet, 107-a gland gas outlet, 108, 109-a sealing ring micro-cavity hole, 110-a handheld structure, 501-a chip liquid inlet hole, 502-a chip gas outlet hole, 503-a chip liquid inlet micro-channel, 504-a chip liquid outlet micro-channel, 505-a chip serpentine tube amplification cavity, 506-a chip glass layer, 507-a chip silicon sheet layer and 510-a heat conducting sheet. The left and right sides of the mounting groove are defined by taking the hole side direction of the mounting groove as a vector direction in the figures 1, 2, 3, 4 and 6.

Detailed Description

The technical solutions in the specific embodiments of the present invention will be clearly and completely described below, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example one

In the embodiment of the present invention shown in fig. 1 and 2, it should be noted that the steps described in the embodiment are not strictly corresponding to the steps described in the content of the present invention.

The embodiment provides a closed microfluidic chip for PCR reaction, which comprises a gland 1, a sealing ring, an elastic part layer and a chip 5, wherein the gland 1, the sealing ring and the elastic part layer are arranged from top to bottom; the gland 1 is made of high polymer materials through an injection molding process, one face of the gland 1 is provided with four positioning holes 3, and the four positioning holes 3 are respectively positioned at the upper left corner, the upper right corner, the lower left corner and the lower right corner of the gland 1.

Two iron sheets 2 are arranged on one surface of the gland 1 in an embedding mode in the injection molding process, and the two iron sheets 2 are symmetrically arranged on the left side and the right side of the gland 1; two handheld structures 110 are symmetrically arranged on the left side and the right side of the gland 1. The gland 1 is further designed with a buckle structure, in this embodiment, a mounting groove hole side buckle 105 is arranged on the gland 1 plane on the hole side of each mounting groove 9, two left side buckles 101 and 102 are arranged on the gland 1 plane on the left side of each mounting groove 9, and two right side buckles 103 and 104 are arranged on the gland 1 plane on the right side of each mounting groove 9. In other embodiments, the number of the iron sheets 2 may be four, and the four iron sheets 2 are symmetrically placed on two sides of the gland 1; the hole side bayonets 105 of each mounting groove 9 may be two, and the snap structure of the left and right sides of each mounting groove 9 may be three. Of course, the iron sheet 2 may be replaced by other metal having ferromagnetism in other embodiments, and two nickel sheets are symmetrically embedded into two sides of the gland 1 in one embodiment. Four mounting grooves 9 are formed on one side of the gland 1 during the injection molding process in this embodiment. In other embodiments, it is preferable that the cover 1 is provided with more than four mounting grooves 9.

A gland sample inlet 106 and a gland gas outlet 107 are also arranged on the gland 1, a sealing ring 6 containing a microcavity hole is vertically arranged at the gland sample inlet 106 and the gland gas outlet 107, and a buffering silica gel gasket 7 is arranged at the opposite side of the sealing ring 6 containing the microcavity hole. In this embodiment, the sealing rings 6 with micro-cavity holes are two silica gel gaskets with micro-cavity holes, and the thickness of the buffer silica gel gasket 7 is 0.1 mm. The microcavity holes 108 and 109 of the two sealing rings are used for sample communication and gas communication. In this embodiment, there are four gland sample inlets 106 and four gland gas outlets 107 in total, there are eight sealing rings 6 with microcavity holes and four buffering silica gel gaskets 7 in total, and the eight sealing rings 6 with microcavity holes and the four buffering silica gel gaskets 7 constitute the sealing ring and the elastic member layer in the present invention. In other embodiments, the number of the elastic members, that is, the buffer silica gel pads 7 in this embodiment, may be two, and the elastic members may also be made of other materials with elasticity, and the thickness thereof may also be 0.11 mm.

Four chips 5 are assembled on the gland 1, and each chip 5 is fixed in any one mounting groove 9 of the gland 1 through two mounting groove left side buckles 101 and 102, two mounting groove right side buckles 103 and 104 and one mounting groove hole side buckle 105. In other embodiments, the number of the assembled chips 5 may vary with the number of the mounting grooves 9, and the number of the chips 5 may be less than or equal to the number of the mounting grooves 9 on the gland 1.

As shown in fig. 7, 8 and 9, the chip 5 is provided with a glass layer 506 of the chip, a silicon layer 507 of the chip and a heat conductive sheet 510 from top to bottom. One side of the chip 5 is provided with a chip liquid inlet 501, a chip gas outlet 502, a chip liquid inlet microchannel 503 and a chip liquid outlet microchannel 504. A chip serpentine amplification chamber 505 is provided in the chip layer 507 by an 8 "wafer etching process. In other embodiments, two or more of the chip serpentine amplification chambers 505 may be disposed in the chip layer 507. Preferably, four serpentine amplification chambers 505 are disposed in the silicon layer 507 of the chip.

In the present embodiment, the thickness of the thermally conductive sheet 510 is 0.05 mm. Preferably, the thermally conductive sheet 510 is a graphite sheet. In other embodiments, the thickness of the heat conducting sheet 510 may be 0.1mm, and may also be 0.55mm, and the heat conducting sheet is a brass sheet.

In other embodiments, four mounting grooves 9 are formed on the gland 1, four chips 5 are assembled, and a silicon layer 507 of each chip 5 is provided with a serpentine amplification cavity 505 of four chips, so that detection of sixteen nucleic acid samples can be completed at one time, and the nucleic acid detection efficiency is effectively improved.

As shown in FIG. 7, each chip 5 is provided with a chip inlet 501, a chip inlet microchannel 503, a chip serpentine amplification chamber 505, a chip outlet microchannel 504 and a chip outlet 502, which are connected in sequence. The sample enters the coiled pipe amplification cavity 505 of the chip through the liquid inlet hole 501 of the chip through the liquid inlet micro-channel 503 of the chip through the gland sample inlet 106 and the micro-cavity hole 108 of the sealing ring for amplification reaction, and the redundant gas is discharged from the gas outlet hole 502 of the chip through the liquid outlet micro-channel 504 of the chip and the micro-cavity hole 109 of the sealing ring and finally from the gland gas outlet 107. In other embodiments, the number of inlet holes 501, inlet microchannels 503, outlet microchannels 504 and outlet holes 502 of each chip varies with the number of serpentine amplification chambers 505 of the chip. In some embodiments, each chip 5 is provided with four serpentine amplification chambers 505, and accordingly each chip 5 is provided with four inlet holes 501, four inlet microchannels 503, four outlet microchannels 504 and four outlet holes 502, and the inlet microchannel 503, the serpentine amplification chambers 505 and the outlet microchannels 504 are sequentially connected to form four chambers without communicating with each other.

Further, when the sample addition is completed, the sealing film 4 disposed on the other side of the cap 1 is covered on each of the cap inlet port 106 and the cap outlet port 107 in this embodiment, thereby completing the sealing of the chip 5 and forming a closed PCR reaction environment. In this example, a biocompatible rubber film was used as the sealing film, and the plastic film was resistant to a temperature of 100 ℃ for 1 hour. Of course, in other embodiments, the sealing film is made of biocompatible TPE, and the plastic film can resist the temperature of 100 ℃ for 1.5 hours.

In this embodiment, through with four locating holes 3 with chip and the accurate location of outside heat transfer seat 8, can realize that outside heat passes through heat transfer seat 8 and conducts heat for conducting strip 510, conducting strip 510 gives the silicon chip layer 507 of chip with the heat again to effectively realize that the rise and fall temperature of the coiled pipe amplification chamber 505 of chip and the rise and fall temperature of heat transfer seat 8 are almost same frequency, and the average rise and fall temperature can be realized to more than 15 ℃/s, can reduce 40 PCR amplification cycles in the past market consuming time 60min to 5 min.

The mounting groove 9 is also a fluorescence detection area of an external device, and when the sample completes the amplification reaction in the coiled tube amplification cavity 505 in the chip 5, the sample can be subjected to fluorescence detection by the external device.

The above description of the embodiments is only intended to facilitate the understanding of the method and the core idea of the present invention. It should be noted that, for those skilled in the art, it is possible to make various improvements and modifications to the present invention without departing from the principle of the present invention, and those improvements and modifications also fall within the scope of the claims of the present invention.

Claims (15)

1. A closed microfluidic chip for PCR reaction is characterized in that: the device comprises a gland and at least one chip, wherein one side of the gland is provided with at least one mounting groove, a buckle, at least one gland sample inlet and at least one gland gas outlet, and the other side of the gland is provided with a sealing film; the chip is installed in the installation groove through the buckle; a sealing ring containing a micro-cavity hole is arranged between the chip and the gland, the micro-cavity hole is a liquid inlet hole and an air outlet hole, the liquid inlet hole is communicated with the sample inlet of the gland, and the air outlet hole is communicated with the air outlet of the gland; the sealing film covers each gland sample inlet and gland gas outlet.

2. The closed microfluidic chip for PCR reaction according to claim 1, wherein: ferromagnetic metal is embedded into the pressure cover through an injection molding process, and the ferromagnetic metal interacts with a magnet on external equipment to generate attraction.

3. The closed microfluidic chip for PCR reaction according to claim 1, wherein: the sealing film is made of a biocompatible and malleable material.

4. The closed microfluidic chip for PCR reaction according to claim 1, wherein: the chip is provided with a first structure body, a second structure body and a heat conducting fin from top to bottom, wherein the first structure body and the second structure body are both of sheet structures.

5. The closed microfluidic chip for PCR reaction according to claim 4, wherein: the heat conducting sheet is made of a heat conducting material with the heat conducting coefficient not lower than 50W/(m.K).

6. The closed microfluidic chip for PCR reaction according to claim 4, wherein: the second structure body is provided with at least one amplification cavity, at least one liquid inlet micro-channel and at least one liquid outlet micro-channel.

7. The closed microfluidic chip for PCR reaction according to claim 1, wherein: an elastic part is further arranged between the chip and the gland, the elastic part is arranged on the other side of the sealing ring with the micro-cavity hole, and the elastic part and the sealing ring with the micro-cavity hole enable the chip to be horizontally arranged on the gland.

8. The closed microfluidic chip for PCR reaction according to claim 1, wherein: the gland is provided with a positioning hole, and the gland is positioned on external equipment through the positioning hole.

9. The closed microfluidic chip for PCR reaction according to any one of claims 1 to 8, wherein: the mounting groove is provided with four, every the assembly has a slice in the mounting groove the chip, every the chip is provided with an amplification chamber.

10. The closed microfluidic chip for PCR reaction according to claim 4, wherein: the first structure body is a glass sheet, and the second structure body is a silicon sheet.

11. The closed microfluidic chip for PCR reaction according to claim 1, wherein: and hand-held structures are symmetrically arranged on two sides of the gland.

12. The closed microfluidic chip for PCR reaction according to claim 2, wherein: the ferromagnetic metal is iron sheets, and the two iron sheets are symmetrically arranged on two sides of the gland.

13. The closed microfluidic chip for PCR reaction according to claim 6, wherein: the amplification cavity is a coiled pipe amplification cavity.

14. The closed microfluidic chip for PCR reaction according to claim 3, wherein: the material also has temperature resistance, and the temperature resistance time of the material at 100 ℃ is more than 0.5 h.

15. The closed microfluidic chip for PCR reaction according to claim 5, wherein: the thickness of the heat conducting sheet is 0.05mm-0.2 mm.

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