CN212770649U - Centrifugal micro-fluidic chip integrating nucleic acid extraction and fluorescence amplification - Google Patents
Centrifugal micro-fluidic chip integrating nucleic acid extraction and fluorescence amplification Download PDFInfo
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- CN212770649U CN212770649U CN202021071424.4U CN202021071424U CN212770649U CN 212770649 U CN212770649 U CN 212770649U CN 202021071424 U CN202021071424 U CN 202021071424U CN 212770649 U CN212770649 U CN 212770649U
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Abstract
The utility model discloses a centrifugal micro-fluidic chip integrating nucleic acid extraction and fluorescence amplification, which comprises a device base and a chip substrate, wherein the upper end of the device base is fixedly connected with an upper mounting plate through a support rod, the left side of the upper mounting plate is fixedly connected with a laser PCR detection module through a crank connecting rod, the middle part of the inner cavity of the upper mounting plate is fixedly connected with an electromagnet coil, the electromagnet coil is electrically connected with a control component, the middle part of the upper end of the device base is fixedly connected with a centrifugal motor, the tail end of an output shaft of the centrifugal motor is fixedly connected with a driving shaft, the driving shaft penetrates through the upper mounting plate and is fixedly connected with the chip mounting base, the middle part of the inner cavity of the chip mounting base is fixedly connected with an electric heating plate, the upper end of the chip mounting base is fixedly, the chip has rich functions, is practical and has certain popularization value.
Description
Technical Field
The utility model relates to a nucleic acid detects technical field, specifically is a centrifugal micro-fluidic chip of integrated nucleic acid extraction and fluorescence amplification.
Background
The nucleic acid detection has the characteristics of high sensitivity and good specificity, and occupies an important position in life science and medical detection. The detection of nucleic acid requires a series of complicated nucleic acid extraction steps to obtain a high-purity nucleic acid template. The conventional manual extraction of nucleic acid has the defects of low efficiency, easy error and the like. The large-scale automation equipment has the defects of large volume, high price, high reagent consumption, need of professional technical background personnel for operation, need of building a special detection laboratory and the like, and is inconvenient to popularize in basic medical institutions.
The micro-fluidic chip technology is a new technology for accurately controlling and controlling nano-liter and pico-liter fluid (biological sample fluid) in a flow channel with a micron scale, and can integrate basic operation units of sample preparation, reaction, separation, detection, cell culture, sorting, cracking and the like in the fields of chemistry, biology and the like or a chip with a few square centimeters (even smaller) by applying the technology, and a network is formed by a micro-flow channel so as to control the fluid to penetrate through the whole system and replace a technical platform with various functions of a conventional chemical or biological laboratory. The basic characteristic and the greatest advantage of the microfluidic chip laboratory are that various unit technologies are flexibly combined and integrated on a small platform with controllable whole. The micro-fluidic chip provides an ideal automatic platform for a plurality of steps of sample flowing, sample mixing, nucleic acid extraction, waste liquid treatment, nucleic acid PCR amplification, fluorescence detection and the like related to sample treatment and nucleic acid amplification detection through a controllable micro-fluidic network formed by a micro valve, a micro pump and a micro channel.
Therefore, a centrifugal microfluidic chip integrating nucleic acid extraction and fluorescence amplification is provided.
SUMMERY OF THE UTILITY MODEL
An object of the utility model is to provide an integrated nucleic acid draws centrifugal micro-fluidic chip with fluorescence amplification to solve the problem that proposes in the above-mentioned background art.
In order to achieve the above object, the utility model provides a following technical scheme: a centrifugal microfluidic chip integrating nucleic acid extraction and fluorescence amplification comprises a device base and a chip substrate, wherein the left end of the device base is fixedly connected with a level gauge, the lower end of the device base is fixedly connected with a spiral supporting leg, the right end of the device base is fixedly connected with a control panel through a rotating shaft, the upper end of the device base is fixedly connected with an upper mounting plate through a supporting rod, the left side of the upper mounting plate is fixedly connected with a laser PCR detection module through a crank connecting rod, the middle part of the inner cavity of the upper mounting plate is fixedly connected with an electromagnet coil, the electromagnet coil is electrically connected with a control assembly, the middle part of the upper end of the device base is fixedly connected with a centrifugal motor, the tail end of an output shaft of the centrifugal motor is fixedly connected with a driving shaft, the driving shaft penetrates through the upper, the chip mounting base is fixedly connected with a chip buckle, the upper end of the chip mounting base is connected with a chip substrate through a chip buckle movable joint, the chip substrate is a fan-shaped substrate and is provided with an injection sample cracking groove close to a round part, the right part of the upper end of the injection sample cracking groove is in seamless connection with an injection sample adding hole, a sealing layer is fixedly connected in the injection sample adding hole, the bottom of an inner cavity of the injection sample cracking groove is fixedly connected with a packaging clamping groove if intervention is performed, a magnetic particle ball and a non-magnetic particle ball are movably clamped in the packaging clamping groove, the front end of the injection sample cracking groove is in seamless connection with a drain valve, the outlet end of the drain valve is in seamless connection with a nucleic acid quantitative groove, the inlet end of the nucleic acid quantitative groove is in seamless connection with a first siphon flow channel, the inlet end of the first siphon flow channel is in seamless connection with, the outer side of the annular flow channel is seamlessly connected with a plurality of nucleic acid amplification holes along the radius direction of the chip substrate, the tail end of the nucleic acid quantifying groove is seamlessly connected with a first waste liquid groove, the tail end of the annular flow channel is seamlessly connected with a second waste liquid groove, the first waste liquid groove and the second waste liquid groove are seamlessly connected through a communicating pipe, and the communicating pipe is further provided with an air vent.
Furthermore, the spirit level is the bubble spirit level, the spiral stabilizer blade is equipped with three at least and is the annular array, control panel up end fixedly connected with liquid crystal display and USB socket, the USB socket is equipped with a plurality ofly.
Furthermore, the electromagnet coil is an annular coil, and the control assembly is fixedly connected to the left side of the lower end face of the upper mounting plate.
Furthermore, the ceramic panel is fixedly connected to the upper end face of the electric heating plate, and a heat insulation layer is filled between the electric heating plate and the chip mounting base.
Further, the first waste liquid tank, the nucleic acid diluting tank and the annular flow path are connected to the vent hole through minute pipes.
Furthermore, the surface of the drain valve material is required to be subjected to hydrophobic modification treatment, and the surfaces of the first siphon flow channel and the second siphon flow channel material are subjected to hydrophilic modification treatment.
Furthermore, a diluent liquid sac is pre-packaged in the nucleic acid dilution tank, and PCR reaction reagent dry powder is pre-packaged in the nucleic acid amplification hole.
Compared with the prior art, the beneficial effects of the utility model are that:
this kind of centrifugal micro-fluidic chip of integrated nucleic acid extraction and fluorescence amplification structural design is reasonable, convenient to use, and this chip drive power only needs a centrifugal motor, need not fluid drive sources such as external syringe pump, peristaltic pump, vacuum pump, simple structure, small in size, and the whole system's of being convenient for realization is miniaturized and integrates. The steps of cell lysis, quantification, DNA/RNA extraction, nucleic acid dilution and purification, PCR amplification, fluorescence detection and the like are integrated on the microfluidic chip, so that pollution and errors caused by manual operation are avoided, the time length of molecular detection is greatly shortened, and the detection sensitivity and precision are improved.
Drawings
Fig. 1 is a cross-sectional view of the present invention;
FIG. 2 is a top view of the chip mounting base of the present invention;
fig. 3 is a top view of the control panel of the present invention;
FIG. 4 is a partial cross-sectional view of the chip mounting base of the present invention
Fig. 5 is a top view of the chip substrate of the present invention;
fig. 6 is a schematic structural view of the sample-adding cracking tank of the present invention.
In the figure: 1 device base, 11 level meter, 12 spiral support leg, 13 rotation shaft, 14 control panel, 15 liquid crystal display screen, 16USB socket, 17 support rod, 18 upper portion mounting plate, 2 electromagnet coil, 21 control component, 22 centrifugal motor, 23 drive shaft, 3 chip mounting base, 31 electric heating plate, 32 ceramic panel, 33 heat-insulating layer, 34 chip buckle, 36 crank connecting rod, 4 laser PCR detection module, 5 chip base plate, 51 nucleic acid amplification hole, 52 annular flow channel, 53 second siphon flow channel, 54 nucleic acid dilution tank, 55 first siphon flow channel, 56 nucleic acid quantification tank, 57 first waste liquid tank, 58 second waste liquid tank, 59 vent hole, 50 trap valve, 6 sample adding cracking tank, 61 injection sample adding hole, 62 sealing layer, 63 prepackaging connection tube, 64 nonmagnetic particle ball, 65 magnetic particle ball, 66.
Detailed Description
The technical solutions in the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative work belong to the protection scope of the present invention.
Referring to fig. 1-6, the present invention provides a technical solution: a centrifugal micro-fluidic chip integrating nucleic acid extraction and fluorescence amplification comprises a device base 1 and a chip substrate 5, wherein the left end of the device base 1 is fixedly connected with a level 11, the level is a bubble level, the lower end of the device base 1 is fixedly connected with a spiral supporting leg 12, the spiral supporting leg 12 is at least provided with three spiral supporting legs and is in an annular array, the right end of the device base 1 is fixedly connected with a control panel 14 through a rotating shaft 13, the upper end surface of the control panel 14 is fixedly connected with a liquid crystal display screen 15 and a USB socket 16, the USB sockets 16 are provided with a plurality of spiral supporting legs, the upper end of the device base 1 is fixedly connected with an upper mounting plate 18 through a supporting rod 17, the left side of the upper mounting plate 18 is fixedly connected with a laser PCR detection module 4 through a crank connecting rod 36, the laser PCR detection module 4 comprises a laser generation assembly and a fluorescence receiving assembly, the electromagnetic coil 2 is an annular coil, the electromagnetic coil 2 is electrically connected with a control component 21, the control component 21 is fixedly connected to the left side of the lower end face of the upper mounting plate 18, the middle part of the upper end of the device base 1 is fixedly connected with a centrifugal motor 22, the tail end of an output shaft of the centrifugal motor 22 is fixedly connected with a driving shaft 23, the driving shaft 23 penetrates through the upper mounting plate 18 and is fixedly connected with a chip mounting base 3, the middle part of an inner cavity of the chip mounting base 3 is fixedly connected with an electric heating plate 31, the upper end face of the electric heating plate 31 is fixedly connected with a ceramic panel 32, a heat insulation layer 33 is filled between the electric heating plate 31 and the chip mounting base 3, the upper end of the chip mounting base 3 is fixedly connected with a chip buckle 34, the upper end of the chip mounting base 3 is movably clamped with a chip base plate 5 through the chip buckle 34, the right part of the upper end of the sample adding cracking tank 6 is seamlessly connected with an injection sample adding hole 61, a sealing layer 62 is fixedly connected in the injection sample adding hole 61, the bottom of the inner cavity of the sample adding cracking tank 6 is fixedly connected with a plurality of interference packaging clamping grooves 63, the pre-packaging clamping grooves 63 are movably clamped with magnetic particle balls 65 and non-magnetic particle balls 64, the front end of the sample adding cracking tank 6 is seamlessly connected with a drain valve 50, the outlet end of the drain valve 50 is seamlessly connected with a nucleic acid quantifying tank 56, the inlet end of the nucleic acid quantifying tank 56 is seamlessly connected with a first siphon runner 55, the inlet end of the first siphon runner 55 is seamlessly connected with the outlet end of a nucleic acid diluting tank 54, the inlet end of the nucleic acid diluting tank 54 is seamlessly connected with an annular runner 52 through a second siphon runner 53, and the outer side of the annular runner 52 is seamlessly connected with, the tail end of the nucleic acid quantifying groove 56 is seamlessly connected with a first waste liquid groove 57, the tail end of the annular flow passage 52 is seamlessly connected with a second waste liquid groove 58, the first waste liquid groove 57 and the second waste liquid groove 58 are seamlessly connected through a communicating pipe 66, the communicating pipe 66 is also provided with a vent hole 59, the first waste liquid groove 57, the nucleic acid diluting groove 54 and the annular flow passage 52 are all connected with the vent hole 59 through micro pipelines, the surface of the material of the drain valve 50 needs to be subjected to hydrophobic modification treatment, the surfaces of the materials of the first siphon flow passage 55 and the second siphon flow passage 53 are subjected to hydrophilic modification treatment, a diluting liquid sac is packaged in the nucleic acid diluting groove 54 in advance, the formula of the diluting liquid is A, the formula A contains guanidine isothiocyanate, a surfactant, BSA, gelatin and other components, a PCR reaction reagent dry powder is packaged in the nucleic acid amplifying hole 51 in advance, and the formula of the dry powder is B, the formula B comprises components such as enzyme, dNTPs, primers and the like required by PCR reaction.
The working principle is as follows:
1. adding a sample to be detected into the sample adding cracking slot 6 through the injection sample adding hole 61;
2. placing the microfluidic chip substrate 5 on the chip mounting base 3 and fixing the microfluidic chip substrate by the chip fastener 34;
3. starting the control assembly 21 and the electromagnet coil 2 to generate a variable magnetic field, wherein the chip substrate 5 is static, and under the action of the variable magnetic field, the magnetic particle balls 65 move back and forth along the radial direction and interact with the non-magnetic particle balls 64 pre-packaged in the sample groove 6 to complete the processes of homogenizing the sample to be detected and cracking cells;
4. the centrifugal motor 22 starts to rotate, when the rotating speed of the centrifugal motor 22 reaches a certain speed, the drain valve 50 is broken through, the sample in the sample adding and cracking groove 6 enters the nucleic acid quantifying groove 56, cell debris, impurities and the like generated in the sample homogenizing and cell cracking process adhere to the wall under the action of centrifugal force, the enrichment of nucleic acid, protein and the like in the sample in the supernatant close to the center of the circle is detected, the nucleic acid quantification is completed, and at the moment, the first siphon flow channel 55 is in a closed state under the action of the centrifugal force;
5. the centrifugal motor 22 stops rotating, and the siphon runner 55 is opened;
6. the centrifugal motor 22 is started to rotate again, and the nucleic acid sample to be detected which is quantified in the nucleic acid quantifying groove 56 is thrown into the nucleic acid diluting groove 54;
7. the centrifugal motor 22 performs rapid acceleration and slow deceleration process circulation and high-temperature heating on the dilution mixing area, and at the moment, the diluent in the diluent sac is released, so that the target DNA in the sample to be detected is cracked, and the dilution process is completed;
8. the centrifugal motor 22 stops rotating, and the second siphon flow passage 53 is opened;
9. the centrifugal motor 22 is started to rotate, and the diluted sample to be detected enters the nucleic acid amplification hole;
10. the electric heating plate 31 carries out alternate heating circulation required by PCR amplification on the nucleic acid amplification holes 51 to finish PCR amplification, the laser PCR detection module 4 is used for carrying out laser irradiation on the nucleic acid amplification holes 51, and the laser PCR detection module can scan and read out the light intensity of each fluorescence channel in each PCR amplification cavity to draw a QPCR curve.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (7)
1. A centrifugal micro-fluidic chip integrating nucleic acid extraction and fluorescence amplification comprises a device base (1) and a chip substrate (5), and is characterized in that: the device comprises a device base (1), a level gauge (11) is fixedly connected to the left end of the device base (1), a spiral supporting leg (12) is fixedly connected to the lower end of the device base (1), a control panel (14) is fixedly connected to the right end of the device base (1) through a rotating shaft (13), an upper mounting plate (18) is fixedly connected to the upper end of the device base (1) through a supporting rod (17), a laser PCR detection module (4) is fixedly connected to the left side of the upper mounting plate (18) through a crank connecting rod (36), an electromagnet coil (2) is fixedly connected to the middle of an inner cavity of the upper mounting plate (18), a control assembly (21) is electrically connected to the electromagnet coil (2), a centrifugal motor (22) is fixedly connected to the middle of the upper end of the device base (1), a driving shaft (23) is fixedly connected to the tail end of an output shaft of the centrifugal motor (22), and, chip mounting base (3) inner chamber middle part fixedly connected with electric plate (31), chip mounting base (3) upper end fixedly connected with chip buckle (34), chip mounting base (3) upper end has chip base plate (5) through chip buckle (34) activity joint, chip base plate (5) are fan-shaped base plate and are close to circular department and offered application of sample schizolysis groove (6), application of sample schizolysis groove (6) upper end right part seamless connection has injection application of sample hole (61), fixedly connected with sealing layer (62) in injection application of sample hole (61), application of sample schizolysis groove (6) inner chamber bottom fixedly connected with if intervene encapsulation draw-in groove (63), activity joint has magnetic particle ball (65) and non-magnetic particle ball (64) in prepackage draw-in groove (63), application of sample schizolysis groove (6) front end seamless connection has trap (50), the outlet end of the drain valve (50) is seamlessly connected with a nucleic acid quantification groove (56), the inlet end of the nucleic acid quantification groove (56) is seamlessly connected with a first siphon flow channel (55), the inlet end of the first siphon flow passage (55) is seamlessly connected with the outlet end of the nucleic acid diluting tank (54), the inlet end of the nucleic acid diluting well (54) is seamlessly connected with an annular flow passage (52) through a second siphon flow passage (53), the outer side of the annular flow passage (52) is seamlessly connected with a plurality of nucleic acid amplification holes (51) along the radius direction of the chip substrate (5), the end of the nucleic acid quantitative groove (56) is seamlessly connected with a first waste liquid groove (57), the tail end of the annular flow passage (52) is connected with a second waste liquid groove (58) in a seamless mode, the first waste liquid tank (57) and the second waste liquid tank (58) are connected in a seamless mode through a communication pipe (66), and a vent hole (59) is further formed in the communication pipe (66).
2. The centrifugal microfluidic chip integrating nucleic acid extraction and fluorescence amplification as claimed in claim 1, wherein: the spirit level is the bubble spirit level, spiral stabilizer blade (12) are equipped with three at least and are the annular array, control panel (14) up end fixedly connected with liquid crystal display (15) and USB socket (16), USB socket (16) are equipped with a plurality ofly.
3. The centrifugal microfluidic chip integrating nucleic acid extraction and fluorescence amplification as claimed in claim 1, wherein: the electromagnet coil (2) is an annular coil, and the control assembly (21) is fixedly connected to the left side of the lower end face of the upper mounting plate (18).
4. The centrifugal microfluidic chip integrating nucleic acid extraction and fluorescence amplification as claimed in claim 1, wherein: the ceramic panel (32) is fixedly connected to the upper end face of the electric heating plate (31), and a heat insulation layer (33) is filled between the electric heating plate (31) and the chip mounting base (3).
5. The centrifugal microfluidic chip integrating nucleic acid extraction and fluorescence amplification as claimed in claim 1, wherein: the first waste liquid tank (57), the nucleic acid diluting tank (54) and the annular flow channel (52) are connected with the vent hole (59) through a micro pipeline.
6. The centrifugal microfluidic chip integrating nucleic acid extraction and fluorescence amplification as claimed in claim 1, wherein: the surface of the material of the drain valve (50) needs to be subjected to hydrophobic modification treatment, and the surfaces of the material of the first siphon runner (55) and the second siphon runner (53) are subjected to hydrophilic modification treatment.
7. The centrifugal microfluidic chip integrating nucleic acid extraction and fluorescence amplification as claimed in claim 1, wherein: the nucleic acid dilution tank (54) is internally pre-packaged with a diluent sac, and the nucleic acid amplification hole (51) is pre-packaged with PCR reaction reagent dry powder.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113322175A (en) * | 2021-07-01 | 2021-08-31 | 清华大学深圳国际研究生院 | Real-time fluorescence constant temperature nucleic acid amplification detection device |
| CN114509575A (en) * | 2022-04-19 | 2022-05-17 | 天津德祥生物技术有限公司 | Microfluidic detection device |
-
2020
- 2020-06-11 CN CN202021071424.4U patent/CN212770649U/en active Active
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113322175A (en) * | 2021-07-01 | 2021-08-31 | 清华大学深圳国际研究生院 | Real-time fluorescence constant temperature nucleic acid amplification detection device |
| CN113322175B (en) * | 2021-07-01 | 2024-01-26 | 清华大学深圳国际研究生院 | Real-time fluorescence constant temperature nucleic acid amplification detection device |
| CN114509575A (en) * | 2022-04-19 | 2022-05-17 | 天津德祥生物技术有限公司 | Microfluidic detection device |
| CN114509575B (en) * | 2022-04-19 | 2022-06-14 | 天津德祥生物技术有限公司 | Microfluidic detection device |
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