CN211057123U - Full-automatic nucleic acid extraction and real-time fluorescence quantitative PCR integrated device - Google Patents
Full-automatic nucleic acid extraction and real-time fluorescence quantitative PCR integrated device Download PDFInfo
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- CN211057123U CN211057123U CN201921786362.2U CN201921786362U CN211057123U CN 211057123 U CN211057123 U CN 211057123U CN 201921786362 U CN201921786362 U CN 201921786362U CN 211057123 U CN211057123 U CN 211057123U
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Abstract
The utility model relates to a full-automatic nucleic acid extraction and real-time fluorescence quantitative PCR integrated device; the full process from uncapping sampling of the blood sampling tube sample tube to timely detection of PCR amplification realizes full-automatic unattended operation, and saves manpower and material resources; the doors of the first buffer area and the second buffer area can not be opened at the same time, and materials are transferred through the first buffer area and the second buffer area and then transferred, so that mutual pollution of all compartments is avoided; assuming that ventilation and air leakage occur, the air flow direction is as follows: the system construction area flows to the nucleic acid extraction area, and reagents placed in the system construction area cannot be polluted if aerosol pollution occurs; the system construction area flows to the amplification area, and if aerosol is polluted in the amplification area, reagents which cannot pollute the system construction area are generated; the outside atmosphere flows to the amplification region, so that even if the amplification region is polluted by aerosol, the aerosol cannot flow out to the air to pollute a laboratory.
Description
Technical Field
The utility model belongs to the technical field of medical instrument, concretely relates to integrative device of full-automatic nucleic acid extraction and real-time fluorescence ration PCR.
Background
In order to realize the anti-pollution function, the existing PCR laboratory is distributed by partition isolation and mainly comprises a nucleic acid extraction chamber, a PCR system construction chamber and a PCR amplification chamber, and PCR materials are conveyed among the chambers through an intermediate isolation channel. Therefore, testers need to be arranged in the middle of each partition room, automation is not realized, the efficiency is not high, and the cost for isolating the partition rooms is very high.
Therefore, it is necessary to develop a fully automatic nucleic acid extraction and real-time fluorescence quantitative PCR integrated device to solve the above problems.
Disclosure of Invention
The utility model aims to solve the above problems and provide an integrated device for full-automatic nucleic acid extraction and real-time fluorescence quantitative PCR.
The utility model discloses a following technical scheme realizes above-mentioned purpose:
a full-automatic nucleic acid extraction and real-time fluorescence quantitative PCR integrated device comprises:
a nucleic acid extraction region for automated extraction of nucleic acids;
an architectonic region for on-demand distribution of PCR reagents into a PCR plate;
the amplification area is used for circularly amplifying the mixed solution of the purified nucleic acid and the PCR reagent at high and low temperatures and recording the change of a fluorescence signal of the device at proper time;
a first buffer area; the nucleic acid extraction area is connected with the system construction area through a first buffer area, and the logistics transfer is carried out through the first buffer area;
a second buffer area; the system construction area is connected with the amplification area through a second buffer area, and the material flow is transmitted through the second buffer area;
the nucleic acid extraction region, the system construction region and the amplification region are all independently arranged, the internal pressure of the nucleic acid extraction region is P1, the internal pressure of the system construction region is P2, the internal pressure of the amplification region is P3, P2> P1> P3, and P3 is negative pressure.
The beneficial effects of the utility model reside in that:
the utility model relates to a full-automatic nucleic acid extraction and real-time fluorescence quantitative PCR integrated device;
1. the full process from uncapping sampling of the blood sampling tube sample tube to timely detection of PCR amplification realizes full-automatic unattended operation, and saves manpower and material resources;
2. the nucleic acid extraction zone, the system construction zone and the amplification zone are designed in the device, and the three compartments are not ventilated with each other and are separated by a first buffer zone and a second buffer zone. The doors of the first buffer area and the second buffer area can not be opened at the same time, and materials are transferred through the first buffer area and the second buffer area and then transferred, so that mutual pollution of all compartments is avoided;
3. samples exposed in the air can not appear in the system construction area and the amplification area, and the PCR tube filled with the samples is sealed, so that the aerosol pollution risk can not be generated;
4. the nucleic acid extraction zone, the system construction zone and the amplification zone are provided with independent ventilation systems, and the internal pressure P1 of the nucleic acid extraction zone, the internal pressure P2 of the system construction zone and the internal pressure of the amplification zone are controlled to be P3; p2> P1> P3, wherein P3 is negative pressure; the purpose of this arrangement is to assume that ventilation and air leakage occur among the three compartments of the nucleic acid extraction zone, the system construction zone and the amplification zone, and the air flow direction is as follows: the system construction area flows to the nucleic acid extraction area, and reagents placed in the system construction area cannot be polluted if aerosol pollution occurs; the system construction area flows to the amplification area, and if aerosol is polluted in the amplification area, reagents which cannot pollute the system construction area are generated; the outside atmosphere flows to the amplification region, so that even if the amplification region is polluted by aerosol, the aerosol cannot flow out to the air to pollute a laboratory.
Drawings
FIG. 1 is a schematic structural diagram of the present application;
FIG. 2 is a schematic diagram of the structure of the nucleic acid extraction zone of the present application;
FIG. 3 is a schematic diagram of the structure of a PCR plate according to the present application;
FIG. 4 is a schematic diagram of the architecture of the architectural region of the present application;
FIG. 5 is a schematic diagram of the structure of an amplification region in the present application.
Detailed Description
The present invention will be further explained with reference to the accompanying drawings:
as shown in FIG. 1, a full-automatic nucleic acid extraction and real-time fluorescence quantitative PCR integrated device comprises:
a nucleic acid extraction region 100 for automatic extraction of nucleic acid;
an architecture region 200 for dispensing PCR reagents on demand into a PCR plate;
an amplification area 300 for performing high and low temperature cyclic amplification on the mixture of the purified nucleic acid and the PCR reagent and recording the change of the fluorescence signal of the device in time;
a first buffer 400; the nucleic acid extraction zone 100 is connected with the system construction zone 200 through a first buffer zone 400, and is in logistics transmission through the first buffer zone 400;
a second buffer 500; the system construction area 200 is connected with the amplification area 300 through a second buffer area 500, and the material flow is transmitted through the second buffer area 500;
the nucleic acid extraction region 100, the architecture region 200 and the amplification region 300 are all separately arranged, and the internal pressure of the nucleic acid extraction region 100 is P1, the internal pressure of the architecture region 200 is P2, the internal pressure of the amplification region 300 is P3, P2> P1> P3, wherein P3 is negative pressure.
As shown in FIG. 1, each of the first buffer area 400 and the second buffer area includes at least two doors, a transfer device, and when the transfer device transfers the product in the nucleic acid extraction area 100 to the system construction area 200, initially, all the doors are closed; thirdly, at least one door close to the system construction area 200 is kept for isolating the nucleic acid extraction area 100 from the system construction area 200, and the product is transferred to the first buffer area 400 by the transfer device; thirdly, at least one door close to the nucleic acid extraction zone 100 is kept closed for isolating the nucleic acid extraction zone 100 from the system construction zone 200; the transfer device transfers the product into the system construction zone 200 with all doors closed;
when the transfer device transfers the products in the system construction region 200 to the amplification region 300, initially, all the doors are closed; thirdly, reserving at least one door close to the amplification zone 300 for isolating the system construction zone 200 and the amplification zone 300, and transferring the product to the second buffer zone 500 by the transfer equipment; third, at least one door close to the system construction zone 200 is kept closed for isolating the system construction zone 200 from the amplification zone 300; the transfer device transfers the product to the amplification zone 300, all doors closed.
In some embodiments, the first buffer 400 includes gate a401, gate B402;
the second buffer 500 includes a gate C501, a gate D502;
when the transfer device transfers the products in the nucleic acid extraction region 100 to the system construction region 200, initially, all the doors are closed; thirdly, the door B402 is closed for isolating the nucleic acid extraction zone 100 from the system construction zone 200, the door A401 is opened, and the transfer device transfers the product into the first buffer zone 400; again, gate A401 is closed, isolating the nucleic acid extraction zone 100 from the architecture zone 200; door B402 is opened, the transfer device transfers the product into the system building area 200, and all doors are closed;
when the transfer device transfers the products in the system construction region 200 to the amplification region 300, initially, all the doors are closed; thirdly, the door D502 is closed and used for isolating the system building area 200 and the amplification area 300, the door C501 is opened, and the transfer equipment transfers the products to the second buffer area 500; third, gate C501 is closed, isolating the architecture construction zone 200 from the amplification zone 300; door D502 is opened and the transfer device transfers product to the amplification zone 300, all doors are closed.
The nucleic acid extraction zone 100 is not vented from the construction zone 200 and is separated from the first buffer zone 400. the first buffer zone 400 has two automatically opening and closing doors which cannot be opened at the same time but can be closed at the same time. Opening the gate A401 allows the nucleic acid extraction zone 100 to communicate with the first buffer zone 400; opening door B402 may place the architectural zone 200 in communication with the first buffer zone 400. The PCR plate may be placed back and forth between the nucleic acid extraction zone 100 and the architecture zone 200 through the first buffer zone 400, but the nucleic acid extraction zone 100 and the architecture zone 200 cannot be in direct communication.
The system construction zone 200 is not vented from the amplification zone 300 and is separated from the second buffer zone 500. the second buffer zone 500 has two automatically opening and closing doors that cannot be opened at the same time but can be closed at the same time. Opening door C501 may place the architectural build area 200 in communication with the second buffer area 500; opening gate D502 allows the amplification zone 300 to communicate with the second buffer zone 500. The PCR plate can be placed back and forth between the architecture region 200 and the amplification region 300 through the second buffer region 500, but the architecture region 200 and the amplification region 300 cannot be in direct communication.
The door in the embodiment can be driven by a motor, an air cylinder or an oil cylinder;
in some embodiments, the transfer apparatus in the first buffer zone 400 employs a first clamping jaw 403; the first clamping jaw 403 can be a pneumatic clamping jaw, an electric clamping jaw or an oil cylinder driven clamping jaw;
in some embodiments, the transfer apparatus in the second buffer zone 500 employs a second jaw 503; the second clamping jaw 503 can be a pneumatic clamping jaw, an electric clamping jaw or an oil cylinder driven clamping jaw;
as shown in fig. 2 and 3, the nucleic acid extraction region 100 includes:
an uncapping module 101 for uncovering the sample tube, sucking the sample out by the liquid separating module 105 and then covering the sample tube with the cover; the cover opening module 101 consists of a clamping jaw and a moving assembly and is used for completing actions of clamping and loosening a cover and also performing lifting and descending actions;
a transfer module 102 for transferring the sample tube to the decapping module 101; the transfer module 102 consists of a clamping jaw and a transmission device; the transmission device is belt transmission or screw rod transmission; the sample tube clamping device is used for completing actions of clamping and loosening the sample tube and also performing a moving action of the sample tube;
an extraction module 104 for mixing and oscillating the sample and the extraction reagent to obtain a purified nucleic acid solution; the extraction module 104 consists of a vibration mixing device, a heating device, a magnetic absorption device and a mixing container; the oscillating mixing device drives an eccentric shaft or an eccentric wheel by a motor, and then drives the sample tube to rotate and oscillate by the eccentric shaft or the eccentric wheel; the heat source of the heating device can use a resistance wire or a ceramic heating plate; when the magnetism needs to be absorbed, the electromagnet is electrified or the permanent magnet is close to and attached to the mixing container, and the magnet in the mixing container is absorbed and attached to the pipe wall on one side close to the magnet; the mixing container is used for storing and mixing the sample and the extraction reagent;
a liquid separation module 105 for transferring the sample to the extraction module 104, mixing with the extraction reagent, and distributing the purified nucleic acid to the prepared PCR plate; the liquid separating module 105 consists of a liquid absorbing device and a transmission device; the liquid suction and liquid spray mode of the liquid suction device can be a mode that a motor drives a piston or a mode that a plunger pump sucks liquid, sprays liquid and the like; the transmission mode of the transmission device can be belt transmission, screw rod transmission and the like.
A sealing module 106 for sealing the PCR plate; the film sealing module 106 consists of a stop block, a heat equalizing block, a heat source and a power source; the stopper is used for supporting the PCR plate; the function of the soaking block is to compress the PCR plate and the film and transfer heat to cause the PCR plate and the film to be locally fused together; the heat source can adopt resistance wires or ceramic heating sheets and the like; the power source can be a motor or a cylinder; the film can be sealed by a pressure-sensitive film or a heat-sealing film;
a liquid separation region 107 for placing a PCR plate 801;
an extraction consumable rack 109 for placing extraction consumables;
an extraction reagent rack 111 for holding an extraction reagent.
As shown in fig. 3 and 4, the architecture region 200 includes:
a PCR plate holding block 204 for placing a PCR plate 801;
a PCR reagent holding block 205 for placing PCR reagents;
a PCR reagent separating device 201 for dispensing PCR reagents into the PCR plate 801 according to the requirement; the liquid separating device 201 consists of a liquid absorbing device and a transmission device; the liquid suction and liquid spray mode of the liquid suction device can be a mode that a motor drives a piston or a mode that a plunger pump sucks liquid, sprays liquid and the like; the transmission mode of the transmission device can be belt transmission, screw rod transmission and the like.
A PCR transport device 202 for grasping and transporting the PCR plate 801; the transfer device in the first buffer area 400 is used for transferring the film-sealed PCR plate 801 from the liquid separation area 107 to the PCR conveying device 202; the PCR conveying device 202 consists of a clamping device and a transmission device; the clamping device can be a pneumatic clamping jaw or an electric clamping jaw; the transmission mode of the transmission device can be belt transmission, screw rod transmission and the like.
As shown in fig. 5, the amplification region 300 includes:
a PCR plate transfer means 301 for transferring the PCR plate to the metal bath means 302 or the waste cassette 304; the PCR plate transfer device 301 consists of a clamping device and a transmission device; the clamping device can be a pneumatic clamping jaw or an electric clamping jaw; the transmission mode of the transmission device can be belt transmission, screw rod transmission and the like.
The clamping jaws 503 of the second buffer zone 500 transfer the membrane-enclosed PCR plate 801 from the PCR transport apparatus 202 to the PCR plate transfer apparatus 301.
A metal bath device 302 for controlling the temperature of the mixed solution in the PCR plate 801; the metal bath device consists of a semiconductor refrigeration sheet, a soaking plate and a radiator; the semiconductor refrigerating sheet is used for rapidly heating and cooling; the soaking plate is used for heat transfer; the radiator is used for quickly discharging heat generated by the semiconductor refrigerating sheet;
an optical detection means 303 for detecting a fluorescent signal in the PCR plate 801; the optical detection device 303 is composed of a signal acquisition device, a light filtering device, a transmission device, a light source and the like; the signal acquisition device is used for acquiring a fluorescent signal in the PCR plate 801, and the flow is that a light source obtains light rays with a specific wavelength range after passing through a filter, the light rays with the specific wavelength are used for irradiating a mixture in the PCR plate 801, so that fluorescent substances in the mixture excite fluorescence with the specific wavelength range, and then a camera or a photomultiplier is used for acquiring the fluorescent signal; the filter device is used for filtering the light source to obtain light rays in a specific wavelength range; the light filtering device comprises a plurality of groups of filters for filtering different wavelength ranges; the transmission device is used for switching the filters in different wavelength ranges;
a waste box 304 for collecting the tested PCR plate 801;
as shown in fig. 1-5, the integrated device further comprises:
a frame case 700 for hermetically sealing the integrated device; a sealing door is arranged on the frame shell 700;
the frame casing 700 is provided with a first air inlet 110 at the top of the chamber of the nucleic acid extraction zone 100 and is provided with a first booster fan for blowing air to the inside of the chamber of the nucleic acid extraction zone 100; a first exhaust port 108 is arranged at the back of the chamber of the nucleic acid extraction zone 100 and a first exhaust fan is arranged for exhausting the inside of the chamber of the nucleic acid extraction zone 100; the first booster fan and the first exhaust fan have different air volumes, so that the circulating air flow is formed in the extraction chamber, and the pressure in the chamber is kept at a certain pressure P1;
the frame casing 700 is provided with a second air inlet 206 at the top of the chamber of the system construction area 200 and is provided with a second booster fan for blowing air into the chamber of the system construction area 200; a second exhaust port 203 is arranged at the back of the chamber of the system construction area 200, and a second exhaust fan is arranged for exhausting the gas in the chamber of the system construction area 200; the air volume of the second booster fan is different from that of the second exhaust fan, so that the inside of the chamber of the system construction area 200 forms a circulating air flow and the inside of the chamber is kept at a certain pressure P2;
the frame casing 700 is provided with a third air inlet 306 at the top of the chamber of the amplification zone 300 and is provided with a third booster fan for blowing air into the chamber of the amplification zone 300; a third exhaust port 305 is arranged at the back of the chamber of the amplification zone 300 and is provided with a third exhaust fan for exhausting the inside of the chamber of the amplification zone 300; the third booster fan and the third exhaust fan have different air volumes, so that the circulating air flow is formed in the extraction chamber and the pressure in the chamber is kept at a certain pressure P3;
the frame case 700 is provided with an exhaust port 701 communicating with the first exhaust port 108, the second exhaust port 203, and the third exhaust port 305, respectively.
In some embodiments, a butt joint is provided at the end of off-gas port 701 for butting against the off-gas duct of the laboratory.
In some embodiments, a filter for filtering contaminants is disposed in front of each of the first exhaust fan, the second exhaust fan, and the third exhaust fan.
The specific implementation mode of nucleic acid extraction in the application is as follows:
1) the transferring module 102 transfers the blood collection tube sample tube to the uncovering module 101;
2) the uncovering module 101 uncovers the cover of the blood collection tube sample tube;
3) the liquid separation module 105 absorbs the sample in the blood collection tube sample tube and transfers the sample into the extraction consumable material in the extraction module 105 for extraction treatment;
4) after the nucleic acid extraction treatment is finished, the purified nucleic acid solution is sucked by the liquid separating module 105 and transferred into a PCR plate 801 prepared in the liquid separating area 107;
5) the membrane sealing module 106 places the membrane 802 on the PCR plate 801 added with the sample and the PCR reagent, and pushes the membrane 802 and the PCR plate 801 into the membrane sealing module 106 for sealing, and the membrane can be cold-pressed by using a pressure-sensitive membrane or hot-pressed by using a heat-sealing membrane;
6) the membrane sealing module 106 pushes the PCR plate 801 sealed with the membrane out of the liquid separating area 107;
the specific implementation mode of system construction:
1) the PCR reagent liquid separating device 201 dispenses the PCR reagent into the PCR plate 801 as required;
2) the clamping jaw 403 in the first buffer area 400 is used for transferring the PCR plate filled with the PCR reagent to the liquid separation area 107 of the nucleic acid extraction area 100 for preparation;
specific embodiments of the amplification zone 300:
1) the clamping jaws 403 in the first buffer area 400 transfer the film-sealed PCR plate 801 from the liquid separation area 107 to the PCR transport device 202;
2) the clamping jaw 503 of the second buffer area 500 transfers the PCR plate 801 with the sealed film from the PCR conveying device 202 to the PCR plate transfer device 301, and then the PCR plate is transferred to the metal bath device 302 by the PCR transfer device 301 to carry out the PCR amplification process;
3) while the PCR amplification process is carried out, the optical detection device 303 detects the fluorescence signal value in the PCR amplification reaction process in real time and carries out analysis processing by the system;
4) after the PCR amplification process is finished, the PCR plate 801 is taken down from the metal bath device 302 by the PCR plate transfer device 301 and put into the waste material box 304;
implementation of the device linkage:
1) preparation of reagents and consumables: placing a blood collection tube sample tube on the sample containing device 103, placing extraction consumables on the extraction consumable rack 109, placing extraction reagents on the extraction reagent rack 111, placing the sample containing device 103, the extraction consumable rack 109 and the extraction reagent rack 111 into the nucleic acid extraction device 100, and closing the sealing door; the PCR plate 801 is placed in the PCR plate holding block 204, the PCR reagent is placed in the PCR reagent holding block 205, the PCR plate holding block 204 and the PCR reagent holding block 205 are placed in the system construction area 200, and the sealing door is closed.
2) The transferring module 102 transfers the blood collection tube sample tube to the uncovering module 101;
3) the uncovering module 101 uncovers the cover of the blood collection tube sample tube;
4) the liquid separation module 105 absorbs the sample in the blood collection tube sample tube and transfers the sample into the extraction consumable material in the extraction module 104 for extraction treatment;
5) the PCR reagent liquid separating device 201 dispenses the PCR reagent into the PCR plate 801 as required;
6) the first clamping jaw 403 in the first buffer area 400 is used for transferring the PCR plate 801 loaded with the PCR reagents to the liquid separation area 107 of the nucleic acid extraction area 100 for preparation;
7) after the nucleic acid extraction treatment is finished, the extracted nucleic acid solution is sucked by the liquid separation module 105 and transferred into a PCR plate 801 prepared in the liquid separation area 107;
8) the membrane sealing module 106 places the membrane on the PCR plate 801 added with the sample and the PCR reagent, and pushes the membrane sealing module 106 to seal, and the membrane 802 can be cold-pressed by using a pressure-sensitive membrane or hot-pressed by using a heat-sealing membrane;
9) the membrane sealing module 106 pushes out the PCR plate 801 sealed with the membrane;
10) the second clamping jaw 503 of the second buffer area 500 transfers the PCR plate 801 sealed with the film to the PCR plate transfer device 301, and then the PCR plate transfer device 301 transfers the PCR plate 801 to the metal bath device 302 for PCR amplification process;
11) while the PCR amplification process is carried out, the optical detection device 303 timely detects the fluorescence signal value in the PCR amplification reaction process, and the fluorescence signal value is analyzed and processed by the system;
12) after the PCR amplification process is finished, the PCR plate 801 is taken down from the metal bath device 302 by the PCR plate transfer device 301 and put into the waste material box 304;
the protection point of this patent is:
1) the full process from uncapping sampling of the blood sampling tube sample tube to timely detection of PCR amplification realizes full-automatic unattended operation, and saves manpower and material resources;
2) the nucleic acid extraction zone 100, the architecture zone 200, and the amplification zone 300 are designed inside the apparatus, and these three compartments are not ventilated from each other and are separated by the first buffer zone 400 and the second buffer zone 500. The doors of the first buffer area 400 and the second buffer area 500 can not be opened at the same time, and materials are transferred after transferring through the first buffer area 400 and the second buffer area 500, so that mutual pollution of all compartments is avoided;
3) the PCR tube which is exposed in the air and is provided with the sample is not sealed in the system construction area 200 and the amplification area 300, and the aerosol pollution risk is not generated;
4) the nucleic acid extraction zone 100, the architecture zone 200 and the amplification zone 300 are each provided with independent ventilation systems, and the pressure P1 inside the nucleic acid extraction zone 100, the pressure P2 inside the architecture zone 200 and the pressure P3 inside the amplification zone 300 are controlled so that the sizes of P1, P2 and P3 are in the following relationship: p2> P1> P3, wherein P3 is negative pressure.
The foregoing illustrates and describes the principles, general features, and advantages of the present invention. It will be understood by those skilled in the art that the present invention is not limited to the above embodiments, and that the foregoing embodiments and descriptions are provided only to illustrate the principles of the present invention without departing from the spirit and scope of the present invention. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (10)
1. The utility model provides a full-automatic nucleic acid extraction and integrative device of real-time fluorescence quantitative PCR which characterized in that includes:
a nucleic acid extraction region (100) for automatically extracting nucleic acid;
an architecture region (200) for dispensing PCR reagents on demand into a PCR plate;
an amplification area (300) for performing high-low temperature cyclic amplification on the mixture of the purified nucleic acid and the PCR reagent and recording the change of the fluorescence signal of the device in time;
a first buffer (400); the nucleic acid extraction area (100) is connected with the system construction area (200) through a first buffer area (400), and the physical flow transmission is carried out through the first buffer area (400);
a second buffer (500); the system construction area (200) is connected with the amplification area (300) through a second buffer area (500), and the material flow is transmitted through the second buffer area (500);
the nucleic acid extraction region (100), the system construction region (200) and the amplification region (300) are arranged independently, the internal pressure of the nucleic acid extraction region (100) is P1, the internal pressure of the system construction region (200) is P2, the internal pressure of the amplification region (300) is P3, P2> P1> P3, and P3 is negative pressure.
2. The integrated device for full-automatic nucleic acid extraction and real-time fluorescence quantitative PCR according to claim 1,
the first buffer area (400) and the second buffer area (500) comprise at least two doors and transfer equipment; the transfer device is installed inside the first buffer area (400) and the second buffer area (500); the first buffer area (400) is connected with the nucleic acid extraction area (100) through at least one gate, and the first buffer area (400) is connected with the system construction area (200) through at least one gate; the second buffer zone (500) is connected to the architecture zone (200) by at least one gate, and the second buffer zone (500) is connected to the amplification zone (300) by at least one gate.
3. The integrated device for full-automatic nucleic acid extraction and real-time fluorescence quantitative PCR according to claim 2,
the first buffer area (400) comprises a gate A (401) and a gate B (402);
the second buffer area (500) comprises a gate C (501) and a gate D (502);
the first buffer region (400) is connected to the nucleic acid extraction region (100) through a gate A (401), and the first buffer region (400) is connected to the architecture region (200) through a gate B (402); the second buffer (500) is connected to the architecture region (200) through a gate C (501), and the second buffer (500) is connected to the amplification region (300) through a gate D (502).
4. The integrated device for full-automatic nucleic acid extraction and real-time fluorescence quantitative PCR according to claim 3, wherein the gate A (401), the gate B (402), the gate C (501) and the gate D (502) are all gates driven by a motor, a cylinder or an oil cylinder.
5. The integrated device for full-automatic nucleic acid extraction and real-time fluorescence quantitative PCR according to claim 3,
the transfer devices in the first buffer area (400) and the second buffer area (500) are clamping jaws driven by a motor, an air cylinder or an oil cylinder.
6. The integrated device for full-automatic nucleic acid extraction and real-time fluorescence quantitative PCR according to claim 2, wherein the nucleic acid extraction region (100) comprises:
an uncovering module (101) for uncovering the sample tube, sucking the sample out by the liquid-separating module (105) and then covering the sample tube with the cover;
a transfer module (102) for transferring the sample tube to the decapping module (101);
an extraction module (104) for mixing and oscillating the sample and the extraction reagent to obtain a purified nucleic acid solution;
a liquid separation module (105) for transferring the sample to the extraction module (104), mixing with the extraction reagent, and distributing the purified nucleic acid to the prepared PCR plate;
a sealing module (106) for sealing the PCR plate;
a liquid separating area (107) for placing a PCR plate (801);
an extraction consumable rack (109) for placing extraction consumables;
an extraction reagent rack (111) for placing an extraction reagent.
7. The integrated device for full-automatic nucleic acid extraction and real-time fluorescence quantitative PCR according to claim 6, wherein the system building region (200) comprises:
a PCR plate holding means (204) for holding a PCR plate (801);
a PCR reagent containing device (205) for containing a PCR reagent;
a PCR reagent separating device (201) for distributing the PCR reagent into the PCR plate (801) according to the requirement;
a PCR conveying device (202) for grabbing and transporting the PCR plate (801); a transfer device in the first buffer zone (400) is used to transfer the membrane-sealed PCR plate (801) from the liquid separation zone (107) to the PCR transport device (202).
8. The integrated apparatus for full-automatic nucleic acid extraction and real-time fluorescence quantitative PCR according to claim 7, wherein the amplification region (300) comprises:
a PCR plate transfer device (301); the transfer device of the second buffer area (500) is used for transferring the membrane-sealed PCR plate (801) from the PCR transport device (202) to the PCR plate transfer device (301),
a metal bath device (302); transferring the PCR plate to a metal bath device (302) through a PCR transfer device (301) to carry out a PCR amplification process;
an optical detection device (303); while carrying out PCR amplification process, the optical detection device (303) timely detects the fluorescence signal value in the PCR amplification reaction process;
a waste cartridge (304); after the PCR amplification process is completed, the PCR plate 801 is removed from the metal bath device 302 by the PCR plate transfer device 301 and put into the waste box 304.
9. The integrated device for full-automatic nucleic acid extraction and real-time fluorescence quantitative PCR according to any one of claims 1-8, further comprising:
a frame housing (700) for hermetically sealing the integrated device; a sealing door is arranged on the frame shell (700);
the frame shell (700) is provided with a first air inlet (110) at the top of the chamber of the nucleic acid extraction area (100) and is provided with a first booster fan for blowing air to the inside of the chamber of the nucleic acid extraction area (100); a first exhaust port (108) is arranged at the back of the chamber of the nucleic acid extraction area (100) and a first exhaust fan is arranged for exhausting the chamber of the nucleic acid extraction area (100); the first booster fan and the first exhaust fan have different air volumes, so that the circulating air flow is formed in the extraction chamber, and the pressure in the chamber is kept at a certain pressure P1;
the frame shell (700) is provided with a second air inlet (206) at the top of the chamber of the system construction area (200) and is provided with a second booster fan for blowing air into the chamber of the system construction area (200); arranging a second exhaust port (203) at the back of the chamber of the system construction area (200) and installing a second exhaust fan for exhausting the chamber of the system construction area (200); the air quantity of the second booster fan is different from that of the second exhaust fan, so that the inside of the chamber of the system construction area (200) forms circulating air flow and the inside of the chamber is kept at a certain pressure P2;
the frame shell (700) is provided with a third air inlet (306) at the top of the chamber of the amplification area (300) and is provided with a third booster fan for blowing air into the chamber of the amplification area (300); a third exhaust port (305) is arranged at the back of the chamber of the amplification zone (300) and is provided with a third exhaust fan for exhausting the chamber of the amplification zone (300); the third booster fan and the third exhaust fan have different air volumes, so that the circulating air flow is formed in the extraction chamber and the pressure in the chamber is kept at a certain pressure P3;
an exhaust port 701 is provided in the frame case 700 to communicate with the first exhaust port 108, the second exhaust port 203, and the third exhaust port 305, respectively.
10. The integrated apparatus for full-automatic nucleic acid extraction and real-time fluorescence quantitative PCR according to claim 9, wherein a filter for filtering contaminants is disposed in front of each of the first exhaust fan, the second exhaust fan and the third exhaust fan.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201921786362.2U CN211057123U (en) | 2019-10-23 | 2019-10-23 | Full-automatic nucleic acid extraction and real-time fluorescence quantitative PCR integrated device |
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| CN201921786362.2U CN211057123U (en) | 2019-10-23 | 2019-10-23 | Full-automatic nucleic acid extraction and real-time fluorescence quantitative PCR integrated device |
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| CN211057123U true CN211057123U (en) | 2020-07-21 |
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