CN110373312B - Constant-temperature amplification nucleic acid detection device and detection method - Google Patents
Constant-temperature amplification nucleic acid detection device and detection method Download PDFInfo
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- CN110373312B CN110373312B CN201910779673.4A CN201910779673A CN110373312B CN 110373312 B CN110373312 B CN 110373312B CN 201910779673 A CN201910779673 A CN 201910779673A CN 110373312 B CN110373312 B CN 110373312B
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- 238000001514 detection method Methods 0.000 title claims abstract description 69
- 230000003321 amplification Effects 0.000 title claims abstract description 48
- 238000003199 nucleic acid amplification method Methods 0.000 title claims abstract description 48
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 30
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 30
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 30
- 239000007788 liquid Substances 0.000 claims abstract description 34
- 238000000034 method Methods 0.000 claims abstract description 19
- 238000011161 development Methods 0.000 claims abstract description 14
- 230000008569 process Effects 0.000 claims abstract description 13
- 238000012546 transfer Methods 0.000 claims abstract description 10
- 238000010790 dilution Methods 0.000 claims abstract description 8
- 239000012895 dilution Substances 0.000 claims abstract description 8
- 230000008859 change Effects 0.000 claims abstract description 5
- 238000006243 chemical reaction Methods 0.000 claims description 80
- 238000012360 testing method Methods 0.000 claims description 27
- 230000007246 mechanism Effects 0.000 claims description 26
- 238000007789 sealing Methods 0.000 claims description 24
- 238000009396 hybridization Methods 0.000 claims description 10
- 238000001125 extrusion Methods 0.000 claims description 9
- 238000011901 isothermal amplification Methods 0.000 claims description 8
- 238000003825 pressing Methods 0.000 claims description 8
- 239000012528 membrane Substances 0.000 claims description 7
- 239000003085 diluting agent Substances 0.000 claims description 6
- 230000009471 action Effects 0.000 claims description 3
- 238000007865 diluting Methods 0.000 claims description 3
- 238000013021 overheating Methods 0.000 claims description 3
- 238000012545 processing Methods 0.000 claims description 3
- 230000001960 triggered effect Effects 0.000 claims description 3
- 238000004458 analytical method Methods 0.000 abstract description 6
- 238000010438 heat treatment Methods 0.000 abstract description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 10
- 241000463219 Epitheca Species 0.000 description 6
- 239000000463 material Substances 0.000 description 3
- 239000011550 stock solution Substances 0.000 description 3
- 230000007547 defect Effects 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000003752 polymerase chain reaction Methods 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000010191 image analysis Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
Abstract
The invention discloses a constant temperature amplification nucleic acid detection device and a detection method, wherein the nucleic acid detection device comprises: the chip, control by temperature change module, motion module, detection module and control module, control module and motion module, control module and control module are connected. The chip is arranged on the temperature control module and can amplify, dilute and develop a sample; the temperature control module is arranged on the motion module and is used for heating the chip and keeping the chip at a constant temperature; the movement module drives the chip to move to different functional positions and can drive the liquid in the chip to transfer; the detection module can photograph the chip; the control module can receive the pictures of the detection module, process and analyze the pictures and output detection results. The detection device integrates the functions of sample constant temperature amplification, dilution, color development, image recognition and analysis, greatly simplifies the structure of the instrument, reduces the cost, has small volume and improves the portability and operability of the instrument.
Description
Technical Field
The invention relates to the technical field of nucleic acid detection, in particular to a constant-temperature amplification nucleic acid detection device and a detection method.
Background
At present, the molecular diagnosis market is mainly based on various large instruments and equipment and lengthy and complicated tissue sample pretreatment means, most of the devices, such as 3 devices, a nucleic acid extractor, an amplifier and a scanner, are required for completing the detection, and the molecular diagnosis market has the defects of high cost, long time consumption, inaccurate target, low popularity and the like.
The real-time fluorescent quantitative PCR (polymerase chain reaction) is very commonly used in nucleic acid detection, the traditional detection mode is to carry out temperature rise and fall and real-time fluorescent detection on eight-row PCR tubes or 96-well plates, and the whole instrument not only has high power consumption, but also has large volume, complex structure and long detection time.
Therefore, the prior art is not sufficient and needs to be improved.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a detection device and a detection method for nucleic acid amplified at constant temperature.
The technical scheme of the invention is as follows: provided is a nucleic acid detection device for isothermal amplification, comprising: the device comprises a chip, a temperature control module, a motion module, a detection module and a control module, wherein the control module is connected with the motion module, the temperature control module and the detection module.
The chip is arranged on the temperature control module, and the chip can perform amplification reaction of a sample, dilution of an amplified sample and color reaction.
The temperature control module is arranged on the motion module and can heat the chip and keep the chip at a constant temperature.
The motion module can drive the temperature control module to further drive the chip to move to different functional positions, and can drive the liquid in the chip to transfer.
The detection module can photograph the chip.
The control module can receive the pictures of the detection module, process and analyze the pictures and output detection results.
Further, the chip includes: rubber lid, epitheca, main part spare, sealed pad, test paper strip and drain pan, the epitheca with the drain pan lid closes inside a cavity that forms in back, main part spare, sealed pad and the test paper strip all is located in the cavity, sealed pad corresponds the main part spare sets up, be equipped with amplification reaction district and stock solution district on the main part spare, the bottom in stock solution district is equipped with a seal membrane, sealed pad corresponds the stock solution district is equipped with a recess, be equipped with an extrusion passageway in the recess, the extrusion passageway with amplification reaction district UNICOM, the bottom of recess can take place to warp under the exogenic action, the seal membrane is located in the recess, the drain pan corresponds the recess is equipped with a first through-hole, the epitheca corresponds the amplification reaction district is equipped with a second through-hole, the rubber lid is located the epitheca top covers the second through-hole, the one end of test paper strip is fixed sealed pad is kept away from this one end of recess, this one end of main part spare is close to the test paper strip is equipped with a miniflow way, miniflow with amplification reaction district UNICOM, the other end of test paper strip is equipped with the chromogenic reaction district, the epitheca corresponds the tail end of test paper strip is equipped with the identification district.
The temperature control module includes: the chip protection device comprises a heater, a temperature sensor, a chip slot, a temperature overheating protection switch, a chip cover plate and an elastic piece, wherein the chip is inserted into the chip slot, and the elastic piece can clamp the chip.
The motion module includes: driving motor, guiding mechanism and link mechanism, the control by temperature change module is installed on guiding mechanism, driving motor can drive the guiding mechanism motion, and then drives the control by temperature change module motion, link mechanism includes: the middle part of the connecting rod is fixed on the temperature control module through a rotating shaft, the connecting rod comprises a contact end and a free end, the contact end penetrates through a bottom plate of the chip slot and extends into the first through hole to be contacted with the bottom surface of the groove, the connecting rod can be driven by the guide mechanism to rotate around the rotating shaft, the free end can be contacted with the boss in the rotating process, the boss abuts against the free end of the connecting rod, the contact end rotates upwards to press the bottom surface of the groove, the bottom surface of the groove is pressed to be inwards concave to damage the sealing membrane, one end of the reset spring is fixed on the temperature control module, the other end of the reset spring is fixed on the connecting rod, and the connecting rod resets through the reset spring.
The detection module is located directly over the temperature control module, the detection module includes: a light source and a camera assembly.
The control module includes: the control circuit board is connected with the driving motor, the temperature control module, the light source and the microcomputer, and the microcomputer is connected with the camera.
Further, the camera assembly comprises a camera and a lens, wherein the camera is a CCD camera or a COMS camera; the end surface of the boss close to the side of the connecting rod is an inclined plane or a curved surface which inclines downwards; the chip is a micro-fluidic chip, and the identification area is a two-dimensional code.
Further, the mobile terminal comprises a shell, a touch display screen arranged at the top of the shell, a printing module arranged at the rear end of the shell and a movable bin gate arranged on the front side of the shell, wherein the touch display screen and the printing module are connected with a control module, the movable bin gate is installed on the shell through a rotating assembly, and the moving module can drive the chip to jack the bin gate and stretch out the chip.
Further, rotating assembly includes torsional spring and round pin axle, through the drive of torsional spring the door can automatic closure.
The invention also provides a nucleic acid detection method, which comprises the following specific steps:
s1, placing a chip with a sample into a chip slot;
s2, scanning the two-dimensional code of the chip;
s3, carrying out amplification reaction on the sample;
s4, diluting the amplified sample;
s5, carrying out hybridization and color reaction on the amplified sample;
s6, collecting photos in the color development area;
and S7, processing and analyzing the picture and outputting a result.
Further, in the step S2, the temperature control module is driven by the movement module to move the chip to the barcode scanning area, the detection module scans the two-dimensional code of the chip, and sends the two-dimensional code image to the control module; and in the step S3, the temperature control module is controlled by the control module to heat the chip at a constant temperature, and the chip is insulated when the chip is heated to a certain temperature, so that the sample is subjected to amplification reaction.
Further, after amplification is completed in step S4, the motion module drives the temperature control module to drive the chip to move, the free end of the connecting rod abuts against the front end of the boss, the connecting rod is triggered and rotates around the rotating shaft, so that the contact end of the connecting rod rotates upwards to press the bottom of the groove of the sealing pad in the chip, the sealing film at the bottom of the liquid storage tank is damaged, and then the diluent in the liquid storage tank flows out and flows into the amplification reaction region through the pressing channel to dilute the amplified sample.
Further, in the step S5, after the amplified sample is diluted, the motion module continues to move to drive the temperature control module and further drive the chip to continue to move, the free end of the connecting rod moves downward along the inclined surface of the boss, the contact end of the connecting rod continues to rotate upward to continue to press the bottom of the groove, so as to press the pressing channel inside the groove, so that the amplified sample diluted in the amplification reaction region flows into the color development reaction region on the test strip through the micro-channel, and after a period of reaction, hybridization and color development reaction are completed.
Further, in the step S6, after the hybridization and color reaction are completed, the microcomputer controls to turn on the light source, and simultaneously controls the camera to photograph the color reaction area of the chip, so as to obtain the picture of the color reaction area of the chip.
And in the step S7, after the camera shoots the picture of the color reaction area, the microcomputer acquires the picture from the camera, analyzes and processes the picture, and finally displays the detection result through the touch display screen and outputs the result through the printing module.
By adopting the scheme, the detection device integrates the functions of sample constant-temperature amplification, dilution, color development, image identification and analysis, has a compact integral structure and a small volume, and is convenient for detection operation; the chip integrates all reaction systems of the reagents, liquid reagents and the test paper strips are packaged in the shell, active transfer of liquid can be realized, the chip is compact in structure and small in size, the whole reagent reaction testing process is carried out in the chip, the chip consumes materials at one time, and biological risks that internal liquid overflows the detection device can be avoided; the temperature control module reduces the power consumption of the instrument and saves energy; the motion module can realize the motion of chip and the initiative of the interior liquid of chip simultaneously and shift, does not need extra application of sample mechanism, and the wiper mechanism realizes the liquid and shifts, has greatly simplified instrument structure, has reduced the instrument cost, and is small, has improved the portability and the operability of instrument.
Drawings
FIG. 1 is a schematic diagram of an isothermal amplification nucleic acid detection apparatus according to the present invention;
FIG. 2 is a schematic view of the structure of the isothermal amplification nucleic acid detecting apparatus according to the present invention;
FIG. 3 is a schematic view of the internal structure of the isothermal amplification nucleic acid detecting apparatus according to the present invention;
FIG. 4 is a schematic diagram showing the structure of a chip in the isothermal amplification nucleic acid detection apparatus according to the present invention;
FIG. 5 is a sectional view of a chip in the isothermal amplification nucleic acid detecting apparatus according to the present invention;
FIG. 6 is a flow chart of nucleic acid detection according to the present invention.
Detailed Description
The invention is described in detail below with reference to the figures and the specific embodiments.
Referring to FIGS. 1 to 3, the present invention provides an apparatus for detecting nucleic acid amplified at a constant temperature, comprising: chip 1, temperature control module 2, motion module 3, detection module 4 and control module 5, control module 5 with motion module 3, temperature control module 2 and detection module 4 are connected. The chip 1 is arranged on the temperature control module 2, and the chip 1 can perform amplification reaction of a sample, dilution of an amplified sample and color reaction. The temperature control module 2 is arranged on the motion module 3 and can heat the chip 1 and keep the chip 1 at a constant temperature. The motion module 3 can drive the temperature control module 2 to further drive the chip 1 to move to different functional positions, and can drive the transfer of liquid in the chip 1. The detection module 4 can photograph the chip 1. The control module 5 can receive the picture of the detection module 4, process and analyze the picture and output a detection result. Nucleic acid extraction element still includes shell 6, locates the touch-control display screen 7 at 6 tops of shell, locates the print module 8 of 6 rear ends of shell and locate the movable door 9 of 6 front sides of shell, touch-control display screen 7 reach print module 8 all with control module 5 is connected, movable door 9 passes through rotating assembly 10 and installs on the shell 6, motion module 3 can drive 1 backs down of chip door 9 will 1 of stretching out of chip. The rotating assembly 10 comprises a torsion spring and a pin shaft, and the bin door 9 can be automatically closed under the driving of the torsion spring. The constant-temperature amplification nucleic acid detection device integrates the functions of constant-temperature amplification, dilution, color development, image recognition and analysis of samples, and has the advantages of compact integral structure, small volume and convenience in detection operation.
Referring to fig. 4 and 5, the chip 1 includes: rubber cover 11, upper shell 12, main part 13, sealing gasket 14, test paper strip 15 and bottom shell 16. The upper shell 12 and the bottom shell 16 are covered to form a cavity inside, and the main body part 13, the sealing gasket 14 and the test strip 15 are all located in the cavity. The sealing gasket 14 is arranged corresponding to the main body part 13, the main body part 13 is provided with an amplification reaction region 17 and a liquid storage region 18, the liquid storage region 18 is provided with a sealing film 131 corresponding to the bottom of the main body part 13, the sealing film 131 is a layer of film, and the film can be damaged under the extrusion of external force, so that the diluent in the liquid storage region 18 flows out. The sealing pad 14 is provided with a groove 141 corresponding to the liquid storage region 18, an extrusion channel 142 is provided in the groove 141, and the extrusion channel 142 is communicated with the amplification reaction region 17. Specifically, in this embodiment, the sealing pad 14 is a rubber pad, and the bottom of the groove 141 can deform under the action of an external force, so as to press the pressing channel 142. The sealing film 131 is located in the groove 141, the bottom case 16 is provided with a first through hole 161 corresponding to the groove 141, and an external structure can be pressed against the bottom of the groove 141 through the first through hole 161 to press the groove 141. The upper case 12 is provided with a second through hole 121 corresponding to the amplification reaction region 17, the rubber cover 11 is positioned above the upper case 12 and covers the second through hole 121, and a sample can be added to the amplification reaction region 17 through the second through hole 121 by opening the rubber cover 11. One end of the test strip 15 is fixed to the end of the sealing pad 14 away from the groove 141, the end of the main body 13 close to the test strip 15 is provided with a micro-channel 132, the micro-channel 132 is communicated with the amplification reaction region 17, an inlet of the micro-channel 132 is arranged on the side wall of the amplification reaction region 17, and an outlet of the micro-channel 132 is arranged close to the reagent strip 15. The other end of the test strip 15 is provided with a color reaction area 19, the upper shell 12 is provided with a window 122 corresponding to the color reaction area 19, after the amplification reaction is completed, the sample is diluted, and then the bottom of the groove 141 is extruded to extrude the extrusion channel 142, so that the diluted amplification sample in the amplification reaction area 17 flows onto the test strip 15 through the micro flow channel 132, the color reaction is carried out on the color reaction area 19, and finally the final color result can be observed through the window 122. The tail end of epitheca 12 is equipped with identification area 123, specifically, in this embodiment the identification area is the two-dimensional code, through in advance with the two-dimensional code that chip 1 corresponds scans and shoots, uploads to control module, the follow-up discernment chip of being convenient for. The chip 1 integrates all reaction systems of reagents, including sample amplification, dilution and hybridization color development, liquid reagents and test strips are packaged in a shell, and the liquid reagents and the test strips are extruded to extrude the channel 142 and transfer of liquid can be realized through the micro channel 132, the chip 1 is compact in structure and small in size, the whole test process is carried out in the chip 1, the chip 1 is disposable material consumption, and the biological risk that the internal liquid overflows the detection device can be avoided.
Referring to fig. 1 to 3, the temperature control module 2 includes: the chip comprises a heater 21, a temperature sensor 22, a chip slot 23, a temperature overheating protection switch, a chip cover plate 24 and an elastic piece, wherein the chip 1 is inserted into the chip slot 23, and the elastic piece can clamp the chip 1 to prevent the chip 1 from loosening. The temperature control module 2 adopts a silica gel heating film to be matched with a PID temperature control system to realize constant temperature of the chip 1, so that the temperature is more uniform, the temperature control is more accurate, and compared with the conventional PCR amplification, the power consumption of the detection device is reduced, and the energy is saved.
Referring again to fig. 1 to 3, the motion module 3 includes: driving motor 31, guiding mechanism 32 and link mechanism 33, temperature control module 2 is installed on guiding mechanism 32, driving motor 31 can drive guiding mechanism 32 moves, and then drives temperature control module 2 moves, link mechanism includes: the middle part of the connecting rod 33 is fixed on the temperature control module 2 through a rotating shaft 333, the connecting rod 33 comprises a contact end 331 and a free end 332, the contact end 331 penetrates through the bottom plate of the chip slot 23 to be in contact with the bottom surface of the groove 141, the connecting rod 33 can rotate around the rotating shaft 333 under the driving of the guide mechanism 32, the free end 332 can be in contact with the boss 34 in the rotating process, specifically, the end surface of the boss 34 close to the side of the connecting rod 33 is a downward inclined surface or a curved surface, and the top end a of the inclined surface or the curved surface is higher than the bottom end b of the inclined surface or the curved surface. The top end a of the boss 34 abuts against the free end 332 of the connecting rod 33, the contact end 331 rotates upwards and can pass through the first through hole 161 to press the bottom surface of the groove 141, the bottom surface of the groove 141 is pressed and concaved inwards to break the sealing membrane 131, and the diluent in the liquid storage area 18 flows out; the connecting rod 33 further rotates, the free end 332 continues to slide down the inclined surface, and the contact end 331 continues to press the groove 141 and further the pressing channel 142, so that the diluent flows into the amplification reaction zone 17 through the pressing channel 142 to dilute the reaction sample; the connecting rod 33 further rotates, and when the free end reaches the bottom end b of the boss 34, the contact end 331 presses the pressing channel 142, so that the diluted reaction sample in the amplification reaction region 17 flows to the test strip 15 through the micro flow channel 132, and further flows to the color reaction region 19 for color reaction. One end of the reset spring 35 is fixed on the temperature control module 2, the other end of the reset spring is fixed on the connecting rod 33, after the color reaction is completed, the driving motor 31 drives the guide mechanism 32 to drive the temperature control module 2 to retreat and reset, and at the moment, the connecting rod 33 realizes the reset through the direction elasticity of the reset spring 35. The motion module 3 realizes the motion of the chip 1 at different positions through one driving motor 31, the link mechanism 33 can realize the function of actively transferring liquid in the chip 1, and no additional sample adding mechanism or cleaning mechanism is needed to realize the liquid transfer, so that the structure of the detection device is greatly simplified, the cost is reduced, the size is small, and the portability and the operability of the detection device are improved.
Referring to fig. 1 to 3 again, the detection module 4 is disposed right above the temperature control module 2, and the detection module 4 includes: a light source 41 and a camera assembly 42, the camera assembly 42 comprising a camera and a lens, the camera being a CCD camera or a COMS camera. After the color reaction is completed, the control module 5 controls the detection module 4 to photograph the color reaction area 19, and uploads the image to the control module 5 for analysis and result display. The detection module 4 adopts a photographing mode to detect, and is simple and convenient. The control module 5 comprises: the control circuit board is connected with the driving motor 31, the temperature control module 2, the light source and the microcomputer 51, the microcomputer 51 is connected with the camera, the camera shoots the image of the color reaction area 19 and then transmits the image to the microcomputer 51, the microcomputer 51 carries out image analysis and corresponds to a chip two-dimensional code which is collected in advance, then the analysis result is sent to the touch display screen 7 to be displayed, and meanwhile, the result can be printed out through the printing module 8.
Referring to fig. 1 to 6, the present invention further provides a method for detecting nucleic acid, specifically, the method for detecting nucleic acid by using the nucleic acid extracting apparatus in this embodiment includes the following steps:
and S1, putting the chip with the sample into a chip slot.
And S2, scanning the two-dimensional code of the chip, and controlling the movement module to move the chip to the bar code scanning area by the through hole control module to scan the two-dimensional code of the chip.
And step S3, carrying out amplification reaction on the sample. The temperature control module is controlled by the control module to heat the chip at a constant temperature, and the chip is insulated when heated to a certain temperature, so that the sample is subjected to amplification reaction.
And S4, diluting the amplified sample. After the amplification reaction is finished, the movement module drives the temperature control module to further drive the chip to move, the free end of the connecting rod is abutted against the front end of the boss, the connecting rod is triggered to rotate around the rotating shaft, the contact end of the connecting rod rotates upwards to extrude the bottom of the groove of the sealing gasket in the chip, the sealing film at the bottom of the liquid storage tank is damaged, and then diluent in the liquid storage tank flows out and flows into the amplification reaction area through the extrusion channel to dilute and amplify the sample.
And S5, carrying out hybridization and color reaction on the amplified sample. After the amplified sample is diluted, the motion module continues to move to drive the temperature control module and further drive the chip to continue to move, the free end of the connecting rod moves downwards along the inclined plane of the boss, the contact end of the connecting rod continues to rotate upwards to continue to extrude the bottom of the groove, and further extrude the extrusion channel inside the groove, so that the amplified sample diluted in the amplification reaction area flows into the color development reaction area on the test strip through the micro-channel, and after a period of reaction, hybridization and color development reaction are completed.
And S6, carrying out photo collection on the color development area. And after the hybridization and the color reaction are finished, the microcomputer controls to turn on the light source and simultaneously controls the camera to photograph the color reaction area of the chip to obtain the picture of the color reaction area of the chip.
And S7, processing and analyzing the picture and outputting a result. And after the camera shoots the picture of the color development reaction area, the microcomputer acquires the picture from the camera, analyzes and processes the picture, and finally displays the detection result through the touch display screen and outputs the result through the printing module.
In conclusion, the detection device integrates the functions of constant-temperature sample amplification, dilution, color development, image identification and analysis, and has the advantages of compact overall structure, small volume and convenience in detection operation; the chip integrates all reaction systems of the reagents, the liquid reagents and the test paper strips are packaged in the shell, active transfer of liquid can be realized, the chip is compact in structure and small in size, the whole reagent reaction testing process is carried out in the chip, and the chip consumes materials at one time, so that the biological risk that the internal liquid overflows the detection device can be avoided; the temperature control module reduces the power consumption of the instrument and saves energy; the motion module can realize the motion of chip and the initiative transfer of liquid in the chip simultaneously, does not need extra application of sample mechanism, and the wiper mechanism realizes the liquid transfer, has greatly simplified the instrument structure, has reduced the instrument cost, and is small, has improved the portability and the operability of instrument.
The present invention is not limited to the above preferred embodiments, and any modifications, equivalent substitutions and improvements made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. An apparatus for detecting nucleic acid amplified at a constant temperature, comprising: a chip, a temperature control module, a motion module, a detection module and a control module, wherein the control module is connected with the motion module, the temperature control module and the detection module,
the chip is arranged on the temperature control module and can perform amplification reaction of a sample, dilution of an amplified sample and color reaction;
the temperature control module is arranged on the motion module and can heat the chip and keep the chip at a constant temperature;
the motion module can drive the temperature control module to further drive the chip to move to different functional positions and can drive the liquid in the chip to transfer;
the detection module can photograph the chip;
the control module can receive the picture of the detection module, process and analyze the picture and output a detection result;
the chip includes: the device comprises a rubber cover, an upper shell, a main body part, a sealing gasket, a test strip and a bottom shell, wherein a cavity is formed inside the upper shell and the bottom shell after the upper shell and the bottom shell are covered, the main body part, the sealing gasket and the test strip are located in the cavity, the sealing gasket is arranged corresponding to the main body part, an amplification reaction area and a liquid storage area are arranged on the main body part, a sealing membrane is arranged at the bottom of the liquid storage area, the sealing gasket is provided with a groove corresponding to the liquid storage area, an extrusion channel is arranged in the groove and communicated with the amplification reaction area, the bottom of the groove can deform under the action of external force, the sealing membrane is located in the groove, the bottom shell is provided with a first through hole corresponding to the groove, the upper shell is provided with a second through hole corresponding to the amplification reaction area, the rubber cover is located above the upper shell and covers the second through hole, one end of the test strip is fixed at the end, away from the groove, the main body part is provided with a micro-flow channel, the micro-flow channel is communicated with the amplification reaction area, the other end of the test strip is provided with a color development reaction area, the upper shell is provided with a window, and an identification area;
the temperature control module includes: the temperature control device comprises a heater, a temperature sensor, a chip slot, a temperature overheating protection switch, a chip cover plate and an elastic piece, wherein the chip is inserted into the chip slot, and the elastic piece can clamp the chip;
the motion module includes: driving motor, guiding mechanism and link mechanism, the control by temperature change module is installed on guiding mechanism, driving motor can drive the guiding mechanism motion, and then drives the control by temperature change module motion, link mechanism includes: the middle part of the connecting rod is fixed on the temperature control module through a rotating shaft, the connecting rod comprises a contact end and a free end, the contact end penetrates through a bottom plate of the chip slot and extends into the first through hole to be contacted with the bottom surface of the groove, the connecting rod can be driven by the guide mechanism to rotate around the rotating shaft, the free end can be contacted with the boss in the rotating process, the boss abuts against the free end of the connecting rod, the contact end rotates upwards to press the bottom surface of the groove, the bottom surface of the groove is pressed to be inwards concave to damage the sealing membrane, one end of the reset spring is fixed on the temperature control module, the other end of the reset spring is fixed on the connecting rod, and the connecting rod resets through the reset spring.
2. The isothermal amplification nucleic acid detecting device according to claim 1,
the detection module is located directly over the temperature control module, the detection module includes: a light source and a camera assembly;
the control module includes: the control circuit board is connected with the driving motor, the temperature control module, the light source and the microcomputer, and the microcomputer is connected with the camera.
3. The isothermal amplification nucleic acid detection device according to claim 2, wherein the camera assembly comprises a camera and a lens, wherein the camera is a CCD camera or a COMS camera; the end surface of the boss close to the side of the connecting rod is an inclined plane or a curved surface which inclines downwards; the chip is a micro-fluidic chip, and the identification area is a two-dimensional code.
4. The device for detecting the nucleic acid amplification at the constant temperature according to claim 1, further comprising a housing, a touch display screen arranged at the top of the housing, a printing module arranged at the rear end of the housing, and a movable bin gate arranged at the front side of the housing, wherein the touch display screen and the printing module are both connected with the control module, the movable bin gate is mounted on the housing through a rotating component, and the moving module can drive the chip to push the bin gate open so as to extend the chip out.
5. The apparatus according to claim 4, wherein the rotating member comprises a torsion spring and a pin, and the door is driven by the torsion spring to automatically close.
6. A nucleic acid detection method is characterized by comprising the following specific steps:
s1, placing a chip with a sample into a chip slot;
s2, scanning the two-dimensional code of the chip;
s3, carrying out amplification reaction on the sample;
s4, diluting the amplified sample;
s5, carrying out hybridization and color reaction on the amplified sample;
s6, collecting photos in the color development area;
and S7, processing and analyzing the picture and outputting a result.
7. The method for detecting nucleic acid according to claim 6, wherein in step S2, the temperature control module is driven by the motion module to move the chip to the barcode scanning area, the detection module scans the two-dimensional code of the chip and sends the two-dimensional code image to the control module; and in the step S3, the temperature control module is controlled by the control module to heat the chip at a constant temperature, and the chip is insulated when the chip is heated to a certain temperature, so that the sample is subjected to amplification reaction.
8. The method according to claim 6, wherein in step S4, after the amplification is completed, the motion module drives the temperature control module to drive the chip to move, the free end of the rod abuts against the front end of the boss, the rod is triggered and rotates around the rotating shaft, so that the contact end of the rod rotates upward to press the bottom of the groove of the sealing pad in the chip, the sealing film at the bottom of the liquid storage tank is broken, and the diluent in the liquid storage tank flows out and flows into the amplification reaction region through the pressing channel to dilute the amplified sample.
9. The method according to claim 6, wherein in step S5, after the diluted amplified sample is obtained, the motion module continues to move to drive the temperature control module and further drive the chip to continue to move, the free end of the connecting rod moves downwards along the inclined surface of the boss, the contact end of the connecting rod continues to rotate upwards to further press the bottom of the groove and further press the pressing channel inside the groove, so that the amplified sample diluted in the amplification reaction region flows into the chromogenic reaction region on the test strip through the micro-channel, and after a period of reaction, hybridization and chromogenic reaction are completed.
10. The method for detecting nucleic acid according to claim 6, wherein in step S6, after the hybridization and color reaction are completed, the microcomputer controls to turn on the light source and controls the camera to photograph the color reaction area of the chip to obtain the picture of the color reaction area of the chip;
and in the step S7, after the camera captures the picture of the color reaction area, the microcomputer acquires the picture from the camera, analyzes and processes the picture, finally displays the detection result through the touch display screen, and outputs the result through the printing module.
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| CN110305773B (en) * | 2019-08-22 | 2022-11-08 | 深圳市芯思微生物科技有限公司 | Nucleic acid extraction device and extraction method |
| WO2021134511A1 (en) * | 2019-12-31 | 2021-07-08 | 苏州缔因安生物科技有限公司 | Full-integrated small-scale chip type digital pcr detection system and detection method |
| CN113817592A (en) * | 2020-06-19 | 2021-12-21 | 安徽为臻生物工程技术有限公司 | Anti-pollution detection device and method for rapid detection of nucleic acid amplification product and application |
| CN111748463A (en) * | 2020-08-07 | 2020-10-09 | 天津智善生物科技有限公司 | Amplification device applied to nucleic acid detection and detection method based on amplification device |
| CN114317250A (en) * | 2020-09-30 | 2022-04-12 | 富佳生技股份有限公司 | Heating structure, detection chip, nucleic acid detection box and nucleic acid detection equipment |
| CN113324985B (en) * | 2021-06-16 | 2022-03-11 | 博奥生物集团有限公司 | Centrifugal micro-fluidic detection device and centrifugal micro-fluidic detection system |
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| CN113773952A (en) * | 2021-10-12 | 2021-12-10 | 江苏汇先医药技术有限公司 | Handheld LAMP detector and system |
| CN114047170B (en) * | 2021-11-27 | 2022-08-16 | 广州普世君安生物科技有限公司 | Constant temperature fluorescence detector and multichannel fluorescence detection structure |
| CN114100722A (en) * | 2021-12-27 | 2022-03-01 | 北京梓晶生物科技有限公司 | Portable full-automatic nucleic acid constant temperature amplification device |
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