CN113174324A - PCR reaction device - Google Patents
PCR reaction device Download PDFInfo
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- CN113174324A CN113174324A CN202110057507.0A CN202110057507A CN113174324A CN 113174324 A CN113174324 A CN 113174324A CN 202110057507 A CN202110057507 A CN 202110057507A CN 113174324 A CN113174324 A CN 113174324A
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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
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L7/00—Heating or cooling apparatus; Heat insulating devices
- B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/06—Valves, specific forms thereof
Abstract
The invention relates to a PCR reaction device, which comprises a PCR reaction main body, a display module and a power supply device, wherein the display module is arranged on the PCR reaction main body; the PCR reaction main body comprises a nucleic acid extraction device, a nucleic acid purification device, an amplification reaction device and a shell, wherein the nucleic acid extraction device is used for cracking a sample to obtain a mixture; the nucleic acid purification device is used for separating nucleic acid from the mixture; the nucleic acid amplification reaction device is used for amplifying nucleic acid at a specified temperature by using the temperature control unit, and the nucleic acid extraction device, the nucleic acid purification device and the nucleic acid amplification reaction device are all arranged in the shell; the display module is arranged at the end part of the PCR reaction main body and is used for displaying the working states of the nucleic acid extraction device, the nucleic acid purification device and the amplification reaction device; the power supply device is used for supplying power to the PCR reaction main body and the display module, and the power supply device is arranged on the right side of the PCR reaction main body. The integration level of experimental equipment is high, the continuity of experiments is high, and extraction, purification and amplification can be realized on one pipeline layer.
Description
Technical Field
The invention relates to the field of biological detection, in particular to a PCR reaction device.
Background
The extraction, purification and amplification of nucleic acid are routine operations of nucleic acid detection tests and are key steps in the process of gene analysis. In the gene analysis process, specific nucleic acid fragments are often required to be separated and extracted from a mixed sample for subsequent PCR amplification, so that the purification and recovery effects of nucleic acid directly influence the progress and the result of the whole gene analysis process.
The Chinese patent discloses an automatic nucleic acid detection device, which comprises a fixed seat, a main supporting plate vertically fixed on the fixed seat, an automatic sample reaction mechanism and a fixed frame thereof, a nucleic acid detection light path structure and a bearing frame and a controller thereof; the sample reaction automatic mechanism fixing frame is fixed on the main supporting plate, and an outlet of the sample reaction automatic mechanism extends to the lower part of the fixing frame and is positioned outside the fixing frame body; the nucleic acid detection light path structure bearing frame is positioned below the outlet of the sample reaction automatic mechanism and is fixedly connected with the main supporting plate, and the inlet of the nucleic acid detection light path structure is connected with the outlet of the sample reaction automatic mechanism; the controller is arranged on the fixed seat and is respectively and electrically connected with the sample reaction automatic mechanism and the nucleic acid detection light path structure.
Among the above-mentioned technical scheme, on the one hand, detection device requires highly to the environment, and the measuring accuracy is low, and simultaneously, the inside pipeline of chip does not integrate, and reaction solution is difficult to mix and test as expected, and the extraction, purification, the amplification of common nucleic acid need utilize different instruments to accomplish, complex operation.
Disclosure of Invention
Therefore, the PCR reaction device provided by the invention can realize the continuity of experiments, does not need manual intervention in the process, and is convenient and efficient.
In order to achieve the above object, the present invention provides a PCR reaction apparatus comprising: the PCR reaction device comprises a PCR reaction main body, a display module and a power supply device; the PCR reaction main body comprises a nucleic acid extraction device, a nucleic acid purification device, an amplification reaction device and a shell, wherein the nucleic acid extraction device is used for cracking a sample to obtain a mixture of nucleic acid and protein; the nucleic acid purification device is used for separating the nucleic acid from the mixture separately; the nucleic acid amplification reaction device is used for amplifying the nucleic acid at a specified temperature by using an excitation light source, and the nucleic acid extraction device, the nucleic acid purification device and the nucleic acid amplification reaction device are all arranged in the shell; the display module is arranged at the end part of the PCR reaction main body and used for displaying the working states of the nucleic acid extraction device, the nucleic acid purification device and the amplification reaction device; the power supply device is used for supplying power to the PCR reaction main body and the display module, and is arranged on the right side of the PCR reaction main body;
the magnetic control system is characterized by further comprising a central control unit, wherein the central control unit is respectively connected with the ultrasonic unit, the magnetic suction unit and the temperature control unit and is used for controlling the real-time vibration frequency F of the ultrasonic unit, the real-time position L of the magnetic suction unit and the real-time temperature T of the temperature control unit;
a standard reaction matrix R0(F0, L0 and T0) is arranged in the central control unit, wherein F0 represents the standard vibration frequency of the ultrasonic unit, L0 represents the standard position of the magnetic suction unit, and T0 represents the standard temperature of the temperature control unit;
the central control unit controls the ultrasonic unit, the magnetic attraction unit and the temperature control unit according to the standard reaction matrix R0(F0, L0, T0);
a time matrix t (t1, t2, t3) is further arranged in the central control unit, wherein t1 represents the vibration time of the ultrasonic unit, t2 represents the time interval for starting the magnetic suction unit after the ultrasonic unit stops vibrating, and t3 represents the time interval for starting the temperature control unit after the magnetic suction unit stops working;
in the reaction process, if the real-time vibration frequency F of the ultrasonic unit is lower than the standard vibration frequency F0 of the ultrasonic unit, increasing the vibration time t11 of the ultrasonic unit, updating the time matrix t1(t11, t21, t31) in the central control unit, changing the vibration time t11 of the ultrasonic unit to t1(1+ F/F0), changing the time interval for starting the magnetic suction unit to t21 to t2(1-F/F0), and adaptively shortening the time interval of the corresponding temperature control unit to t31 to t3 (1-F/F0);
if the real-time vibration frequency of the ultrasonic unit is higher than or equal to the standard vibration frequency F0 of the ultrasonic unit, maintaining the vibration time t1 of the ultrasonic unit and maintaining the time matrix t (t1, t2, t3) in the central control unit.
Further, the shell comprises an upper shell, a lower shell, an upper cover and a light source cover plate, the nucleic acid extraction device comprises a sample adding layer and a pipeline layer, the nucleic acid purification device comprises an ultrasonic unit and a magnetic absorption unit, the nucleic acid amplification reaction device comprises an excitation light source and a temperature control module, the upper shell is provided with an excitation light source bin and a chip installation bin, the excitation light source bin is used for placing the excitation light source, the chip installation bin is used for placing the pipeline layer and the chip layer, the ultrasonic unit and the magnetic absorption unit are both arranged between the upper cover and the lower cover, and the temperature control module is arranged between the upper cover and the lower cover; the light source cover plate is used for covering the excitation light source bin, and the upper cover is used for covering the chip mounting bin.
Furthermore, the sample adding layer is arranged above the pipeline layer, the ultrasonic unit and the magnetic suction unit are both arranged below the purification bin in the pipeline layer, the excitation light source is arranged on the side surface of the pipeline layer, and the temperature control device module is arranged below the amplification bin in the pipeline layer; the utility model discloses a reagent adding device, including application of sample layer, application of sample layer upside is provided with the application of sample hole for to the internal sample that adds of pipeline, the below of application of sample hole is the application of sample storehouse that a plurality of intervals set up one side of application of sample storehouse still is provided with the pressurization structure, and it includes the pipe wall, is provided with the piston in the pipe wall is inside, the piston along pipe wall reciprocating motion promotes its interior reagent to export outflow or take out to the reagent.
Furthermore, one end of the pressurizing structure is further connected with a motor set, the motor set comprises a connecting piece and a motor, the piston is arranged at the groove of the connecting piece, and the motor drives the connecting piece to rotate so as to drive the piston to move along the pipe wall in a reciprocating mode.
The ultrasonic unit comprises a lower plate, an upper plate, a plurality of guide rails arranged between the upper plate and the lower plate, a spring sleeved on each guide rail to provide reaction force for the upper plate, and an ultrasonic horn penetrating through the upper plate, wherein the lower part of the ultrasonic horn is connected with an ultrasonic transducer through a connecting flange to change the amplitude of the ultrasonic transducer; the ultrasonic amplitude transformer vibrates up and down in the process of transmitting and changing the amplitude; the guide rail is used for limiting the moving direction of the spring when the spring is compressed and ensuring that the ultrasonic transducer is in contact with the purification bin of the pipeline layer.
Further, the magnetic suction unit comprises a support, a magnet and a magnet driving device, wherein the magnet driving device drives the magnet to move on a guide rail on the support to the position below the purification bin.
Furthermore, the temperature control device comprises a heating unit and a heat dissipation unit, the heating unit is a semiconductor refrigeration sheet, the heating unit and the heat dissipation unit are both arranged right below the chip installation bin and are in contact with the chip to be installed, the heat dissipation unit is used for dissipating heat of the amplification bin in the chip, and the heating unit is used for heating the amplification bin in the chip so as to enable the temperature in the amplification bin to be within a preset temperature range; the heat dissipation unit is electrically connected with the control module, and the control module is used for controlling the working states of the heat dissipation unit and the heating unit.
Further, still include the industrial control board, the industrial control board sets up in the display module assembly, the display module assembly is used for showing operating condition, the industrial control board is used for controlling temperature control device, the motor unit the supersound unit and the unit is inhaled to magnetism, display module sets up on the inclined plane of epitheca.
Further, still be provided with the stop valve in the pipeline layer for the break-make to the intraformational corresponding pipeline of pipeline is controlled, stop valve, transfer pump and motor unit cooperation is realized in the intraformational control of pipeline to cracking liquid, washing liquid and eluant, accomplishes corresponding extraction, washing and elution.
Further, the excitation light source is an LED lamp or an LD lamp.
Compared with the prior art, the invention has the advantages that the processes of extraction, purification and amplification of nucleic acid are integrated, and the motor is used for logic control, so that the extraction process of nucleic acid does not need manual intervention, the corresponding purification reaction can be carried out only by controlling the ultrasonic unit and the magnetic attraction unit in the purification process of nucleic acid, and the power supply device carries out whole-process power supply, so that the extraction, purification and amplification processes of nucleic acid are not limited by environment, and the amplification reaction can be completed by controlling the motor and exciting light to work.
Further, ultrasonic energy transfer is to the purification storehouse in, acts on the magnetic bead in the purification storehouse, and the magnetic bead in with the purification storehouse produces small clearance under ultrasonic resonance's effect between magnetic bead and the magnetic bead, and the magnetic substance in the nucleic acid can adsorb on the surface of magnetic bead this moment, and under the vibration effect, the magnetic bead in the purification storehouse does not contact, the absorption of the nucleic acid of being convenient for absorbent more even.
Further, this embodiment integrates the pipeline layer at the chip to complicated experimentation, can control the liquid trend to can improve work efficiency effectively.
Further, after the magnetic unit adsorbs the nucleic acid, the nucleic acid is cleaned, at the moment, the magnet driving device drives the magnet to move to the lower part of the purification bin along the guide rail, and after the cleaning is finished, the nucleic acid is required to be transferred to the amplification bin, the magnet is required to be far away from the purification bin, so that the subsequent transfer of the nucleic acid from the purification bin to the amplification bin is facilitated, and the continuity of the experiment is ensured.
Furthermore, the stop valve is controlled to be opened or closed through the stop valve control panel, so that the on-off of a liquid path on a pipeline layer is controlled, extraction, purification and amplification are realized on the pipeline layer, the continuity of the whole experimental process is guaranteed, and the operation is efficient and rapid.
Drawings
FIG. 1 is an exploded view of a PCR reaction apparatus according to an embodiment of the present invention;
FIG. 2 is a front view of a PCR reaction apparatus according to an embodiment of the present invention;
FIG. 3 is a side view of a PCR reaction apparatus according to an embodiment of the present invention;
FIG. 4 is a rear view of a PCR reaction apparatus according to an embodiment of the present invention;
FIG. 5 is a schematic perspective view of a detection chip for nucleic acid detection according to an embodiment of the present invention;
FIG. 6 is a schematic diagram showing an exploded structure of a detection chip for nucleic acid detection according to an embodiment of the present invention;
fig. 7 is a schematic structural diagram of a sample adding structure according to an embodiment of the present invention.
Detailed Description
In order that the objects and advantages of the invention will be more clearly understood, the invention is further described below with reference to examples; it should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Preferred embodiments of the present invention are described below with reference to the accompanying drawings. It should be understood by those skilled in the art that these embodiments are only for explaining the technical principle of the present invention, and do not limit the scope of the present invention.
It should be noted that in the description of the present invention, the terms of direction or positional relationship indicated by the terms "upper", "lower", "left", "right", "inner", "outer", etc. are based on the directions or positional relationships shown in the drawings, which are only for convenience of description, and do not indicate or imply that the device or element must have a specific orientation, be constructed in a specific orientation, and be operated, and thus, should not be construed as limiting the present invention.
Furthermore, it should be noted that, in the description of the present invention, unless otherwise explicitly specified or limited, the terms "mounted," "connected," and "connected" are to be construed broadly, and may be, for example, fixedly connected, detachably connected, or integrally connected; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meanings of the above terms in the present invention can be understood by those skilled in the art according to specific situations.
Referring to fig. 1 to 4, a PCR reaction apparatus provided in an embodiment of the present invention includes a nucleic acid extracting device 10-1, a nucleic acid purifying device, and a nucleic acid amplification reaction device, wherein the nucleic acid extracting mechanism 10-1, the nucleic acid purifying device, and the nucleic acid amplification reaction device are all disposed in a housing to protect the nucleic acid extracting mechanism 10-1, the nucleic acid purifying device, and the nucleic acid amplification reaction device from damage. The body acid extraction mechanism 10-1 comprises a sample adding device and a pipeline layer, the sample adding device comprises a sample adding layer, a connecting piece and a motor, a plurality of groups of reagent tubes are arranged in the sample adding layer, one ends of the reagent tubes are arranged in grooves of the connecting piece, and the motor controls the connecting piece to rotate so as to drive the reagent tubes to realize liquid pushing and liquid drawing. The sample adding device comprises an ultrasonic unit 90-1 and a magnetic absorption unit 80-1, wherein the ultrasonic unit 90-1 vibrates to scatter magnetic beads in the purification bin, so that the nucleic acid in the mixture is in contact with the magnetic beads more fully, and the magnetic absorption unit 80-1 is used for fixing the nucleic acid and the magnetic beads at the purification bin, at the moment, the motor connected with the first reagent pipe is required to be pulled outwards, the motor connected with the second reagent pipe is pushed inwards, the mixture is further pushed into the purification bin, the mixture is in contact with magnetic beads in the purification bin at the moment, an ultrasonic unit 90-1 and a magnetic suction unit 80-1 are arranged below the pipeline layer, and the ultrasonic unit 90-1 and the magnetic suction unit 80-1 are arranged side by side.
In practical application, in order to prevent the deadweight effect of the ultrasonic unit 90-1 and the magnetic unit 80-1, the embodiment of the invention is further provided with the protective frame 91-1, the protective frame 91-1 is used for protecting the ultrasonic unit 90-1 and the magnetic unit 80-1 in the use process, and meanwhile, the ultrasonic unit and the magnetic unit can be fixed to prevent the ultrasonic unit and the magnetic unit from deviating in the use process. The vibration of the ultrasonic unit 90-1 is used to break up the magnetic beads in the purification chamber, so that the nucleic acid in the mixture is more fully contacted with the magnetic beads, and further, the mixture in the purification chamber needs to be further processed to wash away the proteins and the like in the mixture, and the nucleic acid is retained. Before washing, magnetism unit 80-1 includes support, magnet and magnet drive arrangement, magnet drive arrangement drive magnet is in the guide rail on the support moves to the below in purification storehouse, utilize the attraction relation of magnetic bead and magnet, tightly fix magnetic bead and nucleic acid in purification storehouse department, utilize the third motor of third reagent union coupling to inject the washing liquid inwards to the purification storehouse this moment, it is first reagent pipe and second reagent pipe to need with the cooperation of third reagent pipe, the motor that corresponds can outwards be taken out for impurity such as protein etc. are taken out the purification storehouse, store the waste liquid to application hole or second reagent mouth, in practical application, in order to wash nucleic acid clean, can wash many times, specific process can repeat above-mentioned cleaning process, no longer give unnecessary details here.
Specifically, the infusion pump in the embodiment of the present invention functions to deliver the lysis solution, the cleaning solution, and the eluent to the pipeline layer, and the first reagent tube, the second reagent tube, the third reagent tube, and the fifth reagent tube in the embodiment of the present invention are all infusion pumps.
And finally, in order to separate the nucleic acid adsorbed on the magnetic beads from the magnetic beads, the fifth motor connected with the fifth reagent tube is utilized to inject eluent inwards to the purification bin, and the magnets are arranged at the uppermost end of the support and are in close contact with the purification bin in the cleaning and eluting processes. And (3) introducing the nucleic acid in the purification bin into the amplification bin under the action of a pump valve, and finally closing a double valve connected with the amplification bin to wait for amplification reaction. Then, an excitation light source 20-1 arranged in an excitation light source mechanism bin of the upper shell 40-1 is opened, excitation light with a fixed waveband generated by the excitation light source 20-1 is irradiated on the amplification bin to carry out amplification reaction, the nucleic acid amplification reaction device comprises the excitation light source 20-1, a temperature control module and a lighting module control plate 11-1, the temperature control module is arranged below the pipeline layer amplification bin to control the temperature in the amplification reaction process, and the lighting module control plate 11 receives emergent light of nucleic acid substances in the amplification bin and judges the nucleic acid substances according to collected light signals. The motor group comprises a first motor, a second motor, a third motor, a fourth motor and a fifth motor, the motor group and the magnet driving device are electrically connected with a motor control panel card, control signals of the ultrasonic unit 90-1 and the magnetic attraction unit 80-1 are from the first control panel card 81-1, all control signals are sent from the industrial control panel 100-1, the industrial control panel 100-1 is arranged in the display module 110-1, and the display module 110-1 is arranged on an inclined plane of the end part of the upper shell 40-1 and used for displaying the progress of an experiment and corresponding experiment operation steps. The power supply device 120-1 is used for supplying power to the PCR reaction main body and the display module 110-1, and the power supply device 120-1 is arranged on one side of the PCR reaction main body. The PCR reaction device provided by the embodiment of the invention is provided with the power supply device, realizes ultra-long standby, is convenient to move, is not limited by regions, and can carry out PCR reaction under any environment.
Specifically, the PCR reaction apparatus provided by the embodiment of the present invention further includes a central control unit, wherein the central control unit is respectively connected to the ultrasonic unit, the magnetic attraction unit and the temperature control unit, and is configured to control the real-time vibration frequency F of the ultrasonic unit, the real-time position L of the magnetic attraction unit and the real-time temperature T of the temperature control unit;
a standard reaction matrix R0(F0, L0 and T0) is arranged in the central control unit, wherein F0 represents the standard vibration frequency of the ultrasonic unit, L0 represents the standard position of the magnetic suction unit, and T0 represents the standard temperature of the temperature control unit;
the central control unit controls the ultrasonic unit, the magnetic attraction unit and the temperature control unit according to the standard reaction matrix R0(F0, L0, T0);
a time matrix t (t1, t2, t3) is further arranged in the central control unit, wherein t1 represents the vibration time of the ultrasonic unit, t2 represents the time interval for starting the magnetic suction unit after the ultrasonic unit stops vibrating, and t3 represents the time interval for starting the temperature control unit after the magnetic suction unit stops working;
in the reaction process, if the real-time vibration frequency F of the ultrasonic unit is lower than the standard vibration frequency F0 of the ultrasonic unit, increasing the vibration time t11 of the ultrasonic unit, and updating the time matrix t1(t11, t2, t3) in the central control unit, wherein the vibration time t11 of the ultrasonic unit is t1(1+ F/F0);
if the real-time vibration frequency of the ultrasonic unit is higher than or equal to the standard vibration frequency F0 of the ultrasonic unit, maintaining the vibration time t1 of the ultrasonic unit and maintaining the time matrix t (t1, t2, t3) in the central control unit.
Specifically, the continuous reaction apparatus for nucleic acid extraction, purification and amplification provided by the embodiment of the present invention includes a standard reaction matrix R0(F0, L0 and T0) and a time matrix T (T1, T2 and T3) disposed in a central control unit, where F0 represents a standard vibration frequency of the ultrasonic unit, L0 represents a standard position of the magnetic attraction unit, T0 represents a standard temperature of the temperature control unit, T1 represents a vibration time of the ultrasonic unit, T2 represents a time interval for activating the magnetic attraction unit after the ultrasonic unit stops vibrating, T3 represents a time interval for activating the temperature control unit after the magnetic attraction unit stops vibrating, the vibration time of the ultrasonic unit is controlled according to a real-time frequency of the ultrasonic unit, and if the real-time frequency F of the ultrasonic unit is lower than a standard vibration frequency F0 of the ultrasonic unit, the vibration time T11 of the ultrasonic unit is increased, the vibration time t11 of the ultrasonic unit is t1(1+ F/F0), the real-time vibration frequency and the standard vibration frequency of the ultrasonic unit are compared, and the vibration time of the ultrasonic unit is adjusted according to the comparison result, so that the magnetic beads and nucleic acid substances in the purification bin are fully contacted and adsorbed in the reaction process, and the adsorption effect is better.
Particularly, well accuse unit can set up on the mainboard, can also set up in other positions, as long as can realize with supersound unit, magnetism inhale the unit with the control by temperature change unit be connected can, be convenient for have more the real-time change of the data in the well accuse unit change the vibration time of supersound unit, and magnetism inhale the unit with the start-up time of control by temperature change unit practices thrift the test time, improves experimental efficiency.
In the reaction process, the central control unit controls the vibration frequency of the ultrasonic unit and adjusts the vibration time of the ultrasonic unit according to the relation between the real-time vibration frequency and the standard vibration frequency, if the vibration time of the ultrasonic unit is increased, the time interval for starting the magnetic attraction unit after the ultrasonic unit stops vibrating can be shortened, because the magnetic beads and the nucleic acid substances have better adsorption effect in the ultrasonic vibration process, when the magnetic attraction unit is started to attract the magnetic beads in the amplification bin, the magnetic unit can be started more quickly, the magnetic beads in the amplification bin are adsorbed by the magnet of the magnetic unit, to perform the cleaning process, and save the reaction time, specifically, the time interval for activating the magnetic attraction unit can be changed to t 21-t 2(1-F/F0), the time interval of the corresponding temperature control unit can also be adaptively shortened to t 31-t 3 (1-F/F0).
According to the continuous reaction device based on nucleic acid extraction, purification and amplification, provided by the embodiment of the invention, the vibration time of the ultrasonic unit is changed through the change of the vibration frequency of the ultrasonic unit, and the time interval for starting the magnetic suction unit and the temperature control unit is further controlled according to the vibration time of the ultrasonic unit, so that the experiment progress is accelerated, the time required by the experiment is greatly shortened, and the experiment efficiency is improved.
According to the PCR reaction device provided by the embodiment of the invention, the processes of extraction, purification and amplification of nucleic acid are integrated, and the motor is used for logic control, so that the extraction process of nucleic acid does not need manual intervention, the corresponding purification reaction can be carried out only by controlling the ultrasonic unit and the magnetic attraction unit in the process of nucleic acid purification, and the amplification reaction can be completed by controlling the motor and exciting light to work.
Specifically, the embodiment of the invention also comprises a shell, wherein the nucleic acid extraction device, the nucleic acid purification device and the nucleic acid amplification reaction device are arranged in the shell, the shell comprises an upper shell 40-1, a lower shell 30-1, an upper cover 50-1 and a light source cover plate 60-1, the upper case 40-1 is provided with an excitation light source chamber 42-1 and a chip mounting chamber 41-1, the excitation light source bin 42-1 is used for placing the excitation light source, the chip mounting bin 41-1 is used for placing the pipeline layer and the sample adding layer, the ultrasonic unit 90-1 and the magnetic attraction unit 80-1 are both arranged between the upper shell 40-1 and the lower shell 30-1, the temperature control device is disposed between the upper case 40-1 and the lower case 30-1; the light source cover plate 60-1 is used for covering the excitation light source chamber 42-1, and the upper cover 50-1 is used for covering the chip mounting chamber 41-1. The nucleic acid extraction device 10-1, the nucleic acid purification device and the nucleic acid amplification reaction device are all arranged in the shell, so that the internal structure of the shell is effectively protected, in addition, in the experimental process, a certain amount of heat can be generated by an excitation light source in the upper shell 40-1 in the working process, so that the temperature of the excitation light source bin 42-1 is influenced, and optionally, an air inlet fan 31-1 is arranged below the excitation light source bin 42-1, so that the heat in the excitation light source bin 42-1 is released.
Specifically, the nucleic acid extraction device 10-1 comprises a sample adding layer and a pipeline layer, the sample adding layer and the pipeline layer are arranged in a chip mounting bin 41-1, a valve control plate 12-1 is further arranged on the nucleic acid extraction device 10-1, a lighting module control plate 11-1 is arranged below an upper cover 50-1, the lighting control plate is used for emitting excitation light to nucleic acid substances in an amplification bin and receiving and collecting fluorescence generated in the amplification bin, the fluorescence is transmitted to an image acquisition device through a glass port in the upper cover 50-1, and the valve control plate 12-1 is used for controlling the on-off of a stop valve on the pipeline layer so as to control the flow direction of liquid in a liquid channel on the pipeline layer. One section of the upper cover 50-1 is provided with a connecting lug which is matched with a connecting hole on the upper shell 40-1 to realize the movable connection of the upper cover 50-1 and the upper shell 40-1. The upper cover 50-1 and the upper shell 40-1 are clamped to fix the nucleic acid extraction device 10-1 in the chip mounting bin 41-1.
Specifically, referring to fig. 5 and 6, the upper side of the sample adding layer 3 is provided with a sample adding hole 302 for adding a sample into the chip, and the sample injected into the chip undergoes nucleic acid extraction, purification, amplification, and reaction. The sample adding layer 3 and the pipeline layer 101 of the present embodiment are movably connected to the limiting frame 106 disposed at the side of the pipeline layer 101 through the clamping strip 304, correspondingly, the inner side of the limiting frame 106 is provided with the first clamping groove 107, and the first clamping groove 107 is connected to each other through the clamping strip 304 in a matching manner, so as to realize the switching and fixing of the relative positions of the sample adding layer 3 and the pipeline layer 101. The switching of relative position indicates the change of the relative distance of application of sample layer 3 and pipeline layer 101 exactly, application of sample layer 3 is by the process of first draw-in groove 107 switching second draw-in groove for the distance between application of sample layer 3 and pipeline layer 101 has become nearly, take out gasket 2 after, application of sample layer 3 and pipeline layer 101 communicate, specifically speaking, gasket 2's primary function is protection application of sample layer 3 and pipeline layer 101 do not communicate, take out the gasket during the use again, wherein, seal membrane 104 is pasted in the downside of pipeline layer 101, in order to realize sealed. The assembled sample adding layer 3, the gasket 2, the pipeline layer 101 and the sealing film 104 form a completely closed whole, and viruses in the sample cannot leak.
As shown in fig. 7, in the embodiment, a second engaging groove is further disposed on a side surface of the limiting frame 106 at the lower side of the first engaging groove 107, the second engaging groove is located at the lower side of the first engaging groove 107, when the sample adding layer 3 is transported or stored, the first card slot 107 is connected with the sample adding layer 3, when the reagent reaction is carried out, the gasket 3 is drawn out, the sample adding layer 3 is pressed downwards, so that the sample adding layer 3 is connected with the second card slot, at the same time, the pricking pin disposed on the pipeline layer 101 punctures the reagent disposed in the sample addition layer 3, so that the reagent and the sample can be mixed and reacted, and the puncture needle is arranged on the pipeline layer 101, and in the practical application process, the pipeline layer 101 is provided with an upright post, the puncture needle is arranged at the circle center of the upright post, the upper end surface of the upright post is oval, and the end surface of the upright post is inclined, so that the puncture needle on the upright post can be conveniently matched with the tail end of the reagent tube, the reagent tube can be smoothly punctured, and the reagent can be filled. According to the detection chip for nucleic acid detection provided by the embodiment of the invention, the first clamping groove 107 and the second clamping groove are arranged, so that the sample adding layer 3 and the pipeline layer 101 can be pressed to generate relative position change after the gasket 2 is extracted, and meanwhile, the pricker punctures a reagent in the sample adding layer 3 to realize sample adding and make a sample and the reagent perform a series of reactions.
As shown in fig. 6, the lower side of the gasket 2 according to the embodiment of the present invention is further provided with a slide rail 202, and correspondingly, the upper side of the pipeline layer 101 is provided with a slide groove 108, and the slide rail 202 is connected with the slide groove 108 in a matching manner, so as to realize the sliding connection between the gasket 2 and the pipeline layer 101. The slide groove 108 of the present embodiment is provided inside the stopper 106 on the pipe layer 101. The end of the gasket 2 is provided with a plurality of notches and protrusions arranged at intervals, wherein the slide rail 202 is arranged on the bottom surface of the outermost protrusion.
As shown in fig. 6, the sample application hole 302 of the present embodiment is provided with a sample application hole cover 303 for sealing. Still set up buckle structure at application of sample layer 3 and pipeline layer 101, be provided with first buckle 301 in one side of application of sample layer 3, the downside of first buckle 301 stretches out the end and stretches out application of sample layer 3's bottom is installing application of sample layer 3 and pipeline layer 101 cooperation back together, through first buckle 301 joint on the side of pipeline layer 101 to prevent application of sample layer 3 and pipeline layer 101 separation.
As shown in fig. 6, the pipeline layer 101 of the present embodiment is provided with two first single valves 102 for controlling the stopping or flowing of the liquid in the pipeline layer 101 during the reaction process; the pipeline layer 101 is further provided with a double valve 103 for cutting off a road for fluid in the pipeline or allowing the fluid to pass through, the double valve 103 is communicated with the amplification bin through the pipeline, and the double valve 103 is used for controlling two ends of the amplification bin to be closed or opened simultaneously so that a closed cavity is formed inside the amplification bin. In fig. 6, handles 201 are further disposed on both sides of the gasket 2 to facilitate extraction of the detection chip for nucleic acid detection. In the embodiment of the invention, the amplification bin is arranged at the edge of the pipeline layer 101, and the amplification bin is of a semi-elliptical structure, so that not only can the reaction of the reaction reagent be realized, but also the convenient positioning and installation can be realized through the convex semi-elliptical structure when in use.
Continuing to refer to fig. 6, in the present embodiment, a row of pricker 105 is disposed on the pipeline layer 101, after the sample-adding layer 3 and the gasket 2 are clamped together, the handle 201 is operated to slide the gasket 2 along the sliding groove 108, when the gasket 2 is pulled out when the gasket slides to a position where the gasket cannot advance, the sample-adding layer 3 is pressed into the second clamping groove from the first clamping groove 107, so that the pricker 105 can prick a reagent tube in the sample-adding layer 3, and further the pricker 105 is communicated with a reagent in the sample-adding layer 3, and when a fluorescence sequence marked in the reagent is complementarily matched with a nucleic acid pricker at a corresponding position, a group of probe sequences with completely complementary sequences is obtained by determining a probe position with the strongest fluorescence intensity. A baffle is disposed outside the needle 105, and it acts as a stop and a positioning function when the sample injection layer 3 is matched with the pipeline layer 101.
Specifically, in the embodiment of the present invention, in the sample loading state, a plurality of sets of reagent tubes are disposed in the sample loading layer 3, the sample loading layer 3 is clamped with the first clamping groove 107 through the clamping strip 304 thereon, in the initial installation state, the sample loading layer 3 is matched with the pipeline layer 101 from top to bottom, and the reagent in the reagent tubes is isolated from the lancet through the gasket 2, so that the lancet and the reagent are prevented from being mixed due to vibration in the transportation process, the sample loading layer and the pipeline layer are protected from being disconnected, and puncturing is avoided. When needs are tested, outwards take gasket 2 out along spout 108, outwards take back along spout 108 for gasket 2 presses application of sample layer 3 downwards for card strip 304 and the second draw-in groove joint on application of sample layer 3, and at this moment, felting needle 105 on setting up on pipeline layer 101 mixes with application of sample layer 3's reagent, introduces reagent into pipeline layer 101 and tests.
Specifically, the gasket structure is arranged, so that the detection chip for detecting nucleic acid can be stored completely in the process of storing and transporting the reagent, and when the detection chip is used, the reagent can be introduced into the pipeline layer only by drawing out the gasket and pressing the sample adding layer downwards.
FIG. 7 is a schematic view of a sample-adding layer according to an embodiment of the present invention; in this embodiment, a sample loading chamber is located below the sample loading hole 302, the sample loading chamber can be connected to a reagent tube for loading a reagent or a sample, a reagent outlet 312 is located at the lower portion of the sample loading chamber, a sealing structure 313 is located between the reagent outlet 312 and the sample loading chamber for sealing, and when a reagent needs to be loaded, the spike 105 can puncture 313 to allow the reagent to enter the fluid pipeline along the reagent outlet 312. A pressurizing structure is arranged on one side of the sample adding bin and comprises a tube wall 305, a piston 308 is arranged in the tube wall 305, and the piston 308 moves towards the sample adding bin to push the reagent in the sample adding bin to flow out to a reagent outlet 312; of course, when the reagent is required to be withdrawn, the piston 308 may also withdraw the reagent or other waste liquid, and a sealing ring 311 is provided at the end of the piston rod of the piston 308 for sealing.
Continuing to refer to fig. 7, the piston rod of this embodiment is further provided with a nut 307, which is in threaded connection with the nut 307 to realize relative rotation, and correspondingly, an output structure, such as an air cylinder and an oil cylinder, is provided at one end of the piston rod, or connected to the piston rod by rotating the output structure, such as a motor and a lead screw, at this time, the piston rod rotates, and only the reagent needs to be pushed to flow out of the reagent outlet. Correspondingly, a guide sleeve 306 is sleeved outside the nut 307, and a corresponding shaft shoulder is arranged inside the pipe wall 305 to position and fix the guide sleeve 306; snap rings 314 are further provided at the outer sides of both ends of the guide sleeve 306 to catch the corresponding guide sleeve 306. A sheath 309 is also provided outside the guide sleeve 306 to protect the piston rod, the nut 307 and the guide sleeve 306. When reagent is injected into the pipeline layer 101, the piston moves toward the sample loading bin to increase the pressure therein, so as to push the reagent to flow toward the reagent outlet 312, thereby realizing reagent injection. With reference to fig. 6, in the embodiment of the present invention, a plurality of sets of reagent tubes are provided, and in the embodiment, five sets of reagent tubes are provided, and different or the same reagents are sequentially applied to the pipeline layer according to the experiment requirement, so that the use efficiency can be greatly improved.
As shown in fig. 7, a second buckle 310 is disposed below the sample adding layer 3, and the second buckle 310 is disposed on a side opposite to the first buckle 301 to prevent the sample adding layer 3 from sliding.
It can be seen that this embodiment is integrated at chip pipeline layer to complicated experimentation, can control the liquid trend to can improve work efficiency effectively.
As can be understood by those skilled in the art, the tube structure disposed on the tube layer is unable to perform related nucleic acid detection tests when the sample adding layer is not connected to the tube layer, so that it is necessary to perform nucleic acid extraction, purification and amplification reactions when the tube layer is in contact with the sample adding layer.
Specifically, the pipeline layer comprises a first sample inlet, a first reagent port, a second reagent port, a third reagent port, a fourth reagent port, a purification bin and an amplification bin, the first sample inlet is connected with the first reagent port through a first pipeline, a first single valve is arranged on the first pipeline, the purification bin comprises an inlet and an outlet, the first sample inlet is connected with the inlet through a second pipeline, the first reagent port is connected with the outlet through a third pipeline, and the second reagent port, the third reagent port and the fourth reagent port are all connected with the inlet through a fourth pipeline; the first end of the amplification bin is provided with a first part of the double valve, the second end of the amplification bin is provided with a second part of the double valve, the first part of the double valve is connected with another single valve through a fifth pipeline, the other single valve is connected with the outlet through a sixth pipeline, and the first part of the double valve and the second part of the double valve are opened and closed simultaneously.
Specifically, the ultrasonic unit 90-1 includes a lower plate, an upper plate, a plurality of guide rails disposed between the upper plate and the lower plate, a spring sleeved on each of the guide rails to provide a reaction force to the upper plate, and an ultrasonic horn penetrating through the upper plate, wherein a lower portion of the ultrasonic horn is connected to an ultrasonic transducer through a connecting flange, and the ultrasonic transducer is disposed between the upper plate and the lower plate to be connected to the ultrasonic transducer to change an amplitude of the transducer. The ultrasonic amplitude transformer vibrates up and down in the process of transmitting and changing the amplitude; the guide rail is used for limiting the moving direction of the spring when the spring is compressed, and ensuring good contact between the ultrasonic transducer and the ultrasonic chip and the effectiveness of ultrasonic energy transfer. In the embodiment of the invention, the ultrasonic chip specifically refers to a purification bin of a pipeline layer. The ultrasonic transducer and the ultrasonic amplitude rod of the embodiment of the invention generate longitudinal and transverse amplitudes in the working process, the upper plate is driven to move for a certain amplitude along with the continuous change of the amplitude, and when the amplitude is large enough, if the limiting rod is still at the preset position, the up-and-down movement of the upper plate and the flange is stopped, so that the amplitude output action is stopped. Therefore, the limiting position of the limiting rod can be adjusted. The lower side plate of the embodiment is provided with a plurality of limiting rods, the upper plate is provided with a plurality of limiting holes matched with the limiting rods and used for the limiting rods to pass through, when the ultrasonic transducer and the ultrasonic amplitude transformer work, the ultrasonic transducer and the ultrasonic amplitude transformer generate vertical amplitude, and the ultrasonic amplitude transformer can move in a preset vertical direction through the limiting rods, so that deviation is avoided; meanwhile, the embodiment adopts a combination mode of the limiting rod and the through hole, so that the deviation of the ultrasonic amplitude transformer in the radial direction, namely the transverse direction, can be avoided, and the limiting rod cannot penetrate through the limiting hole when the deviation in the transverse direction is generated. Ultrasonic energy transfer acts on the magnetic bead in the purification storehouse in to the purification storehouse, and the magnetic bead in with the purification storehouse produces small clearance under the effect of ultrasonic resonance between magnetic bead and the magnetic bead, and the magnetic substance in the nucleic acid can adsorb on the surface of magnetic bead this moment, and under the vibration effect, the magnetic bead contactless in the purification storehouse is convenient for the absorption of nucleic acid for absorbent more even.
Particularly, application of sample layer upside is provided with the application of sample hole for to the internal interpolation sample of pipeline, the below of application of sample hole is the application of sample storehouse that a plurality of intervals set up one side of application of sample storehouse still is provided with the pressurization structure, and it includes the pipe wall, is provided with the piston in the pipe wall is inside, the piston along pipe wall reciprocating motion promotes its interior reagent to export outflow or take out to the reagent. One end of the pressurizing structure is further connected with a motor set, the motor set comprises a connecting piece and a motor, the piston is arranged at the groove of the connecting piece, and the motor drives the connecting piece to rotate so as to drive the piston to move along the pipe wall in a reciprocating mode. The embodiment of the invention also provides a zero reset plate 13-1 which is used for controlling the motor in the motor set to carry out zero reset so as to more accurately control the motor to rotate, realize accurate control on the piston and control the dosage of the reagent.
Specifically, when reagent injection is performed on the pipeline layer, the piston moves towards the sample loading bin to increase the pressure in the sample loading bin so as to push the reagent to flow towards the reagent outlet, and the injection of the reagent is realized. In the embodiment of the invention, a plurality of groups of reagent tubes are arranged, in the embodiment, five groups of reagent tubes are arranged, and different or same reagents are sequentially applied to the pipeline layer according to experiment requirements, so that the use efficiency can be greatly improved. And utilize the motor unit to drive the connecting piece and realize the control to the piston, convenient high efficiency.
It can be seen that in the embodiment, a complex experimental process is integrated on a pipeline layer of a chip, and the trend of liquid can be controlled, so that the working efficiency can be effectively improved.
Specifically, the magnetic unit 80-1 includes a support, a magnet, and a magnet driving device, wherein the magnet driving device drives the magnet to move on a guide rail on the support to a position below the purification bin. In the purification reaction process, in order to prevent nucleic acid from being cleaned by using a cleaning solution, the nucleic acid is cleaned after being adsorbed by the ultrasonic unit, at the moment, the magnet is driven by the magnet driving device to move to the lower part of the purification bin along the guide rail, and when the nucleic acid is required to be transferred to the amplification bin after the cleaning is finished, the magnet is far away from the purification bin, so that the subsequent transfer of the nucleic acid from the purification bin to the amplification bin is facilitated, and the continuity of the experiment is ensured.
Specifically, the temperature control device 70-1 includes a heating unit and a heat dissipation unit, the heating unit is a semiconductor cooling plate, the heating unit and the heat dissipation unit are both disposed under the chip mounting bin 41-1 and are in contact with a chip to be mounted, the heat dissipation unit is configured to dissipate heat from an amplification bin in the chip, and the heating unit is configured to heat the amplification bin in the chip, so that the temperature in the amplification bin is within a preset temperature range; the heat dissipation unit is electrically connected with the control module, and the control module is used for controlling the working states of the heat dissipation unit and the heating unit.
During the experiment, when the amplification reaction is performed, the heating unit of the temperature control device 70-1 starts to perform the first preheating operation; after the chip to be detected is detected again, carrying out second preheating operation on the chip to be detected; heating a chip to be detected; after the chip to be detected reaches the preset detection position, continuously heating the chip to be detected to heat the chip to be detected to the preset reaction temperature; in the heating process, if the temperature of the chip to be detected exceeds a preset detection temperature, starting a heat dissipation unit to perform a first heat dissipation operation; when the chip to be detected reaches the preset detection position and is within the preset detection temperature range, the chip detection device acquires images of the chip to be detected so as to detect the chip. After the reaction is finished, the heat dissipation unit completely dissipates heat for the chip so as to rapidly replace the chip. Through heating unit and radiating element, guarantee to detect that the chip is in suitable temperature range at the amplification reaction in-process, prevent that the temperature from crossing excessively low or too high influence amplification reaction's progress, improve experimental efficiency, guarantee the continuity of experiment. The lower shell 30-1 is further provided with a heat dissipation hole 32-1 and a heat dissipation fan, and in the working process of the PCR reaction device, the air inlet fan 31-1, the heat dissipation fan and the heat dissipation hole 32-1 are arranged below the excitation light source bin 42-1 to form a one-way heat dissipation channel in the PCR reaction device provided by the embodiment of the invention, so that the excitation light source generating heat and the amplification bin can be timely dissipated.
Specifically, the PCR reaction apparatus provided in the embodiment of the present invention further includes a first main board 130-1, a second main board 140-1 and a key board 150-1, wherein the first main board 130-1 is disposed in the lower casing 30-1, and a control circuit for the air intake fan 31-1 and the heat dissipation fan is disposed on the first main board, the heat dissipation fan is disposed on two sidewalls of the lower casing 30, and the heat dissipation hole 30-1 is disposed below the heat dissipation fan. Through holes are formed in the first main board 130-1 and the second main board 140-1, and the temperature control device 70-1 sequentially penetrates through the through holes in the second board card 140-1 and the first board card 130-1, so that the temperature control device 70-1, a heat dissipation fan and a heat dissipation hole are close to each other, and heat dissipation is facilitated. The key board 150-1 is provided with a key for controlling a device corresponding to the key. Specifically, the key may be used to control the power supply device 150-1.
Therefore, the chip detection operation can be rapidly and conveniently carried out by the embodiment. And, through setting up temperature control module in order to heat the chip, through the operating temperature of simulation chip to carry out the detection of chip, not only can improve the detection efficiency of chip, still greatly improved the accuracy when chip detects.
Specifically, in the experimental process, in order to clearly understand the current experimental stage, the device further includes a display module 110-1 and an industrial control board 100-1, the industrial control board 100-1 is disposed in the display module 110-1, the display module 110-1 is used for displaying the working state, the industrial control board 100-1 is used for controlling the temperature control device 70-1, the motor unit, the ultrasonic unit 90-1 and the magnetic unit 80-1, and the display module is disposed on the inclined surface of the upper case. The display module group displays parameters in an experimental stage and an experimental process, so that an experimenter can clearly know the progress in the experimental process, and the experimental process is corrected according to the experimental parameters and an experimental result, the industrial control board 100-1 is connected with the temperature control device 70-1, the motor group, the ultrasonic unit 90-1 and the magnetic suction unit 80-1, the working operation conditions of various devices in the experimental process are integrally grasped, the experimental process is orderly carried out, and the experimental process is continuous and visual.
The pipeline layer is also provided with a pump valve for controlling the on-off of the corresponding pipeline in the pipeline layer, and the pump valve is matched with the motor set to realize the control of the cracking liquid, the cleaning liquid and the eluent in the pipeline layer so as to complete corresponding mixing, cleaning and elution. The pump valve is controlled to be opened or closed through the pump valve control panel, so that the on-off of a liquid path of the pipeline layer is controlled, extraction, purification and amplification are realized on the pipeline layer, the continuity of the whole experimental process is guaranteed, and the operation is efficient and rapid.
The excitation light source is an LED lamp or an LD lamp. The LED lamp or the LD lamp has the advantages of short starting time, high brightness, low energy consumption, small volume, long service life, high safety, low cost and the like, and is widely applied to the field of illumination. And at present, the manufacturing technology of monochromatic high-efficiency LED lamps with various colors is mature and widely applied. The LED light source has low driving voltage (2-5V), small volume and stable output light intensity, and is convenient for miniaturization of an excitation light source device.
So far, the technical solutions of the present invention have been described in connection with the preferred embodiments shown in the drawings, but it is easily understood by those skilled in the art that the scope of the present invention is obviously not limited to these specific embodiments. Equivalent changes or substitutions of related technical features can be made by those skilled in the art without departing from the principle of the invention, and the technical scheme after the changes or substitutions can fall into the protection scope of the invention.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention; various modifications and alterations to this invention will become apparent to those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A PCR reaction apparatus, comprising: the PCR reaction device comprises a PCR reaction main body, a display module and a power supply device; the power supply device is arranged on the side surface of the PCR reaction main body, and the display module is arranged at the end part of the PCR reaction main body;
the PCR reaction main body comprises a nucleic acid extraction device, a nucleic acid purification device, an amplification reaction device and a shell, wherein the nucleic acid extraction device comprises a sample adding layer and a pipeline layer, the sample adding layer is arranged below the pipeline layer, the nucleic acid purification device comprises an ultrasonic unit and a magnetic suction unit, the ultrasonic unit and the magnetic suction unit are arranged below a purification bin in the pipeline layer in parallel, the nucleic acid amplification reaction device comprises a temperature control module, the temperature control module is arranged below the amplification bin in the pipeline layer, and the nucleic acid extraction device is used for cracking a sample to obtain a mixture of nucleic acid and protein; the nucleic acid purification device is used for separating the nucleic acid from the mixture separately; the nucleic acid amplification reaction device is used for amplifying the nucleic acid at a specified temperature by using an excitation light source, and the nucleic acid extraction device, the nucleic acid purification device and the nucleic acid amplification reaction device are all arranged in the shell;
the display module is used for displaying the working states of the nucleic acid extraction device, the nucleic acid purification device and the amplification reaction device, and the power supply device is used for supplying power to the PCR reaction main body and the display module;
the magnetic control system is characterized by further comprising a central control unit, wherein the central control unit is respectively connected with the ultrasonic unit, the magnetic suction unit and the temperature control unit and is used for controlling the real-time vibration frequency F of the ultrasonic unit, the real-time position L of the magnetic suction unit and the real-time temperature T of the temperature control unit;
a standard reaction matrix R0(F0, L0 and T0) is arranged in the central control unit, wherein F0 represents the standard vibration frequency of the ultrasonic unit, L0 represents the standard position of the magnetic suction unit, and T0 represents the standard temperature of the temperature control unit;
the central control unit controls the ultrasonic unit, the magnetic attraction unit and the temperature control unit according to the standard reaction matrix R0(F0, L0, T0);
a time matrix t (t1, t2, t3) is further arranged in the central control unit, wherein t1 represents the vibration time of the ultrasonic unit, t2 represents the time interval for starting the magnetic suction unit after the ultrasonic unit stops vibrating, and t3 represents the time interval for starting the temperature control unit after the magnetic suction unit stops working;
in the reaction process, if the real-time vibration frequency F of the ultrasonic unit is lower than the standard vibration frequency F0 of the ultrasonic unit, increasing the vibration time t11 of the ultrasonic unit, updating the time matrix t1(t11, t21, t31) in the central control unit, wherein the vibration time t11 of the ultrasonic unit is t1(1+ F/F0), the time interval for starting the magnetic suction unit is t 21-t 2(1-F/F0), and the time interval of the temperature control unit is t 31-t 3 (1-F/F0);
if the real-time vibration frequency of the ultrasonic unit is higher than or equal to the standard vibration frequency F0 of the ultrasonic unit, maintaining the vibration time t1 of the ultrasonic unit and maintaining the time matrix t (t1, t2, t3) in the central control unit.
2. The PCR reaction device of claim 1, wherein the nucleic acid amplification reaction device further comprises an optical detection unit, the optical detection unit comprises an excitation light source and a lighting module control board, the housing comprises an upper housing, a lower housing, an upper cover and a light source cover board, the upper housing is provided with an excitation light source chamber and a chip mounting chamber, the excitation light source chamber is used for placing the excitation light source, the chip mounting chamber is used for placing the pipeline layer and the chip layer, the ultrasonic unit and the magnetic unit are both arranged between the upper housing and the lower housing, and the temperature control module is arranged between the upper housing and the lower housing; the light source cover plate is used for covering the excitation light source bin, and the upper cover is used for covering the chip mounting bin.
3. The PCR reaction device of claim 2, wherein the sample adding layer has a sample adding hole on its upper side for adding samples into the pipeline layer, a plurality of sample adding chambers are disposed at intervals under the sample adding hole, and a pressurizing structure is disposed on one side of the sample adding chambers and comprises a tube wall, and a piston is disposed in the tube wall and reciprocates along the tube wall to push the reagents therein to flow out or draw back to the reagent outlet.
4. The PCR reaction device of claim 3, wherein one end of the pressurizing structure is further connected with a motor set, the motor set comprises a connecting piece and a motor, the piston is arranged at the groove of the connecting piece, and the motor drives the connecting piece to rotate so as to drive the piston to reciprocate along the tube wall.
5. The PCR reaction apparatus as claimed in claim 1, wherein the ultrasonic unit comprises a lower plate, an upper plate, a plurality of guide rails disposed between the lower plate and the upper plate, a spring sleeved on each of the guide rails to provide a reaction force to the upper plate, and an ultrasonic horn penetrating through the upper plate, and a lower portion of the ultrasonic horn is connected to an ultrasonic transducer through a connection flange to change an amplitude of the ultrasonic transducer; the ultrasonic amplitude transformer vibrates up and down in the process of transmitting and changing the amplitude; the guide rail is used for limiting the moving direction of the spring when the spring is compressed and ensuring that the ultrasonic transducer is in contact with the purification bin of the pipeline layer.
6. The PCR reaction device of claim 5, wherein the magnetic unit comprises a support, a magnet and a magnet driving device, and the magnet driving device drives the magnet to move on a guide rail on the support to the lower part of the purification bin.
7. The PCR reaction device of claim 6, wherein the temperature control device comprises a heating unit and a heat dissipation unit, the heating unit is a semiconductor cooling plate, the heating unit and the heat dissipation unit are both arranged right below the chip mounting bin and are in contact with the chip to be mounted, the heat dissipation unit is used for dissipating heat of the amplification bin in the chip, and the heating unit is used for heating the amplification bin in the chip so that the temperature in the amplification bin is within a preset temperature range; the heat dissipation unit is electrically connected with the control module, and the control module is used for controlling the working states of the heat dissipation unit and the heating unit.
8. The PCR reaction device of claim 7 further comprising an industrial control board disposed in the display module, the display module is used for displaying the working status, the industrial control board is used for controlling the temperature control device, the motor, the ultrasonic unit and the magnetic unit, and the display module is disposed on the inclined surface of the upper housing.
9. The PCR reaction device of claim 8, wherein a stop valve is further disposed in the pipeline layer for controlling the on-off of the corresponding pipeline in the pipeline layer, and the stop valve, the infusion pump and the motor set cooperate to control the cracking liquid, the cleaning liquid and the eluent in the pipeline layer, so as to complete the corresponding extraction, cleaning and elution.
10. The apparatus for PCR reaction according to claim 2, wherein the excitation light source is an LED lamp or an LD lamp.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120288897A1 (en) * | 2010-02-23 | 2012-11-15 | Genturadx, Inc. | Apparatus and methods for integrated sample preparation, reaction and detection |
| US20160265032A1 (en) * | 2015-03-11 | 2016-09-15 | Hexna Corp. | Portable dna analysis machine |
| CN106916743A (en) * | 2017-03-19 | 2017-07-04 | 北京化工大学 | Integrated nucleic acid extraction and augmentation detection system |
| CN110241008A (en) * | 2019-07-12 | 2019-09-17 | 林艳晶 | A kind of device for extracting and detecting for large biological molecule |
| CN111607506A (en) * | 2020-06-11 | 2020-09-01 | 上海前瞻创新研究院有限公司 | Thin film type nucleic acid amplification micro-fluidic chip and preparation and application method thereof |
-
2021
- 2021-01-15 CN CN202110057507.0A patent/CN113174324B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120288897A1 (en) * | 2010-02-23 | 2012-11-15 | Genturadx, Inc. | Apparatus and methods for integrated sample preparation, reaction and detection |
| US20160265032A1 (en) * | 2015-03-11 | 2016-09-15 | Hexna Corp. | Portable dna analysis machine |
| CN106916743A (en) * | 2017-03-19 | 2017-07-04 | 北京化工大学 | Integrated nucleic acid extraction and augmentation detection system |
| CN110241008A (en) * | 2019-07-12 | 2019-09-17 | 林艳晶 | A kind of device for extracting and detecting for large biological molecule |
| CN111607506A (en) * | 2020-06-11 | 2020-09-01 | 上海前瞻创新研究院有限公司 | Thin film type nucleic acid amplification micro-fluidic chip and preparation and application method thereof |
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Application publication date: 20210727 Assignee: Beijing Linke Biotechnology Co.,Ltd. Assignor: BEIJING INTEGRATED BIOSYSTEMS Co.,Ltd. Contract record no.: X2022990000932 Denomination of invention: PCR reaction device Granted publication date: 20220311 License type: Common License Record date: 20221114 |
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