CN115340946B - Amplification device for nucleic acid detection and detection method based on amplification device - Google Patents
Amplification device for nucleic acid detection and detection method based on amplification device Download PDFInfo
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- CN115340946B CN115340946B CN202211278258.9A CN202211278258A CN115340946B CN 115340946 B CN115340946 B CN 115340946B CN 202211278258 A CN202211278258 A CN 202211278258A CN 115340946 B CN115340946 B CN 115340946B
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Abstract
The invention relates to the technical field of nucleic acid detection, in particular to an amplification device for nucleic acid detection and a detection method based on the amplification device. The amplification device for nucleic acid detection comprises a microfluidic chip, a first temperature control module, a second temperature control module, a mounting rack assembly, a lifting mechanism, an optical detection module and a horizontal moving mechanism. The first side of the micro-fluidic chip is provided with a first sealing film for sealing the first side of the chamber, the second side of the micro-fluidic chip is provided with a second sealing film for sealing the second side of the chamber, the first temperature control module is used for heating the first side of the chamber, the second temperature control module is used for heating the second side of the chamber, and the horizontal moving mechanism drives the optical detection module to horizontally move along the length direction of the micro-fluidic chip so as to sequentially detect samples in the heated chamber in the micro-fluidic chip. Because the first temperature control module and the second temperature control module are arranged, the temperature change treatment can be simultaneously carried out on the first side and the second side of the chamber, and the PCR reaction time can be greatly shortened.
Description
Technical Field
The invention relates to the technical field of nucleic acid detection, in particular to an amplification device for nucleic acid detection and a detection method based on the amplification device.
Background
The molecular diagnosis is carried out by directly detecting a gene sequence, has good specificity and high sensitivity, and is widely applied to the fields of infectious disease diagnosis, genetic gene screening, tumor targeted drug typing and the like. The molecular diagnosis technology comprises molecular hybridization, polymerase Chain Reaction (PCR), gene chip, sequencing and the like, wherein the PCR technology accounts for more than 50 percent and is the standard for diagnosing infectious diseases such as pneumonia and the like.
The existing nucleic acid detection products generally only aim at single or a few pathogens to detect, such as respiratory tract infection, and often only screen a few targets such as influenza A, influenza B and the like, common infectious pathogens further comprise syncytial virus, adenovirus, various coronaviruses, rhinovirus, enterovirus, various parainfluenza virus, chlamydia pneumoniae, mycoplasma, streptococcus and the like, and the multiple nucleic acid detection technology is favorable for more comprehensive and accurate diagnosis, thereby providing a basis for accurate symptomatic treatment.
At present, the following methods are mainly used for detecting multiple nucleic acids:
multi-tube multi-method: one reaction tube detects 1-4 targets, and 12 reaction tubes detect 12-48 targets. Because each reaction tube has different detection targets and needs to be respectively configured with different reaction systems, and each reaction tube needs to be added with the same sample, the operation is complex, the detection time is long, the sample consumption is high, the method depends on a cold chain, and the comprehensive cost is high.
Molecular hybridization method: the reaction system is configured to carry out multiple amplification on a PCR instrument, reaction products are taken out to carry out hybridization, membrane washing and drying are carried out after the hybridization is finished, and then result interpretation is carried out on a fluorescence imaging instrument. But the detection time is long, the whole process needs more than 4 and a half hours at the fastest speed, the operation is complex, the requirement on the professional level of personnel is high, and aerosol pollution is easily caused.
A chip amplification method based on a loop-mediated isothermal amplification technology comprises the following steps: although the detection speed is high and the requirement on the instrument is low, the method cannot be relatively quantitative, can only be used for qualitative detection of an end point, needs cold chain support and has the problem of high false positive rate.
In summary, there is a need for a more accurate, faster, and more convenient multiple nucleic acid detection device in clinical applications.
Disclosure of Invention
Technical problem to be solved
In view of the above disadvantages and shortcomings of the prior art, the present invention provides an amplification device for nucleic acid detection, which has a simple structure and a fast temperature change speed, and can greatly shorten the reaction time of PCR and thus the nucleic acid detection time, and at the same time, can realize PCR detection of more targets.
The detection method based on the amplification device can realize convenient and fast detection.
(II) technical scheme
In order to achieve the purpose, the invention adopts the main technical scheme that:
the present invention provides an amplification device for nucleic acid detection, comprising: the micro-fluidic chip is provided with a plurality of communicated chambers, a first sealing film is arranged on the first side of the micro-fluidic chip to seal the first side of the chambers, and a second sealing film is arranged on the second side of the micro-fluidic chip to seal the second side of the chambers; the first temperature control module is in contact with the first sealing film and used for heating the first side of the cavity; the second temperature control module is in contact with the second sealing film and used for heating the second side of the cavity, and the first temperature control module and the second temperature control module are arranged oppositely up and down; a mounting bracket assembly; the lifting mechanism is arranged on the mounting frame assembly and is connected with the first temperature control module to drive the first temperature control module to move up and down; the optical detection module is used for generating a detection light beam to detect the sample; and the horizontal moving mechanism is connected with the optical detection module and drives the optical detection module to horizontally move along the length direction of the microfluidic chip so as to sequentially detect the samples in the heated chambers in the microfluidic chip.
Preferably, the first temperature control module comprises a first radiator, a first heating plate, a first heat conducting plate and a first heat insulation frame which are connected in sequence; the first heat conducting plate extends out of the first heat insulation frame to contact the first sealing film and heat the first side of the chamber.
Preferably, a first side surface of the first heat insulation frame is concavely provided with a first placing groove for accommodating the first heating plate and the first heat conduction plate; the second side surface of the first heat insulation frame is provided with at least one first through hole, the first through hole is communicated with the first containing groove, the first heat conduction plate is provided with a first protruding part, and the first protruding part extends out of the first through hole; and two sides of the first heat insulation frame extend towards the second side face of the first heat insulation frame to form a light shielding part, the light shielding part is provided with a plurality of detection grooves at intervals, and the detection grooves correspond to the side walls of the inner cavity of the microfluidic chip one by one.
Preferably, the second temperature control module comprises a second radiator, a second heating plate, a second heat conducting plate and a second heat insulation frame which are connected in sequence; the second heat conducting plate extends out of the second heat insulation frame to contact the second sealing film and heat the second side of the chamber.
Preferably, a second placing groove for accommodating the second heating plate and the second heat conducting plate is concavely arranged on the first side surface of the second heat insulation frame; the second side surface of the second heat insulation frame is provided with at least one second through hole, the second through hole is communicated with the second placing groove, the second heat conduction plate is provided with a second protruding part, and the second protruding part extends out of the second through hole; the second side of the second heat insulation frame is also provided with a guide groove along the length direction.
Preferably, the mount assembly comprises a support frame; the lifting mechanism is arranged at the top end of the supporting frame, the first temperature control module is arranged in the first mounting seat, and the lifting mechanism is connected with the first temperature control module through the first mounting seat; the second temperature control module is arranged on the mounting surface through a second mounting seat; the mounting bracket assembly further comprises an auxiliary guide unit, one side of the auxiliary guide unit is connected with the supporting frame, and the other side of the auxiliary guide unit is connected with the first mounting seat.
Preferably, the first temperature control module further comprises a first fan, and the first fan is located on one side of the first radiator and connected with the first mounting seat; the second temperature control module further comprises a second fan, and the second fan is located at one end of the second radiator and connected with the second mounting seat.
Preferably, a stopper is arranged on the second side face of the first heat insulation frame.
Preferably, the auxiliary guide unit comprises a connecting frame arranged on two opposite sides of the supporting frame, a guide rail arranged on the connecting frame, and a sliding block arranged on the first mounting seat, and the sliding block is connected with the guide rail in a sliding manner.
The invention also provides a detection method based on the amplification device, which adopts the amplification device for nucleic acid detection, the amplification device also comprises a controller, the controller is respectively connected with the first temperature control module, the second temperature control module and the optical detection module in a communication way, and the detection method comprises the following steps:
s1, a lifting mechanism drives a first temperature control module to ascend so as to reserve a floating space for placing a microfluidic chip;
s2, placing the microfluidic chip on a second temperature control module, driving the first temperature control module to descend by a lifting mechanism so as to compress the microfluidic chip, and enabling the first temperature control module and the second temperature control module to be in contact with a first sealing film and a second sealing film of the microfluidic chip respectively;
s3, the controller controls the first temperature control module and the second temperature control module to respectively heat the first side and the second side of the cavity in the microfluidic chip;
and S4, the controller controls the horizontal moving mechanism to drive the optical detection module to horizontally move along the length direction of the micro-fluidic chip so as to sequentially detect the samples in the heated chambers in the micro-fluidic chip.
(III) advantageous effects
The beneficial effects of the invention are:
according to the amplification device for nucleic acid detection, the first temperature control module and the second temperature control module are arranged, so that the temperature change treatment can be simultaneously carried out on the first side and the second side of the cavity in the microfluidic chip, the PCR reaction time can be greatly shortened, the nucleic acid detection time is further shortened, the PCR detection of more targets can be realized by matching with a plurality of cavities in the microfluidic chip, and the structure is simple.
The detection method of the amplification device provided by the invention can be used for simultaneously carrying out temperature-changing treatment on the samples in the chambers on the first side and the second side in the chamber in the microfluidic chip, so that the PCR reaction time can be greatly shortened, the nucleic acid detection time is further shortened, and the PCR detection of more targets can be realized by matching with a plurality of chambers in the microfluidic chip.
Drawings
FIG. 1 is a schematic diagram of a first view of an amplification apparatus for nucleic acid detection according to the present invention;
FIG. 2 is a schematic diagram of a second perspective view of an amplification apparatus for nucleic acid detection according to the present invention;
FIG. 3 is a schematic diagram of a third perspective view of an amplification apparatus for nucleic acid detection according to the present invention (the mounting frame assembly is not shown);
FIG. 4 is a schematic cross-sectional view of an optical detection module;
FIG. 5 is a schematic diagram of the operation of the optical detection module;
FIG. 6 is a schematic structural diagram of a first temperature control module (first fan not shown);
FIG. 7 is an exploded view of FIG. 6;
FIG. 8 is a schematic view of a first insulating frame;
FIG. 9 is a schematic view of a second temperature control module (second fan not shown)
FIG. 10 is an exploded view of FIG. 9;
FIG. 11 is a schematic view of a second insulating frame.
[ instruction of reference ]
1: a microfluidic chip;
2: a first temperature control module; 21: a first heat sink; 22: a first heating plate; 23: a first heat-conducting plate; 231: a first projecting portion; 24: a first heat insulating frame; 241: a first through hole; 242: a light shielding portion; 2421: detecting a groove; 243: a stopper portion; 25: a first fan;
3: a second temperature control module; 31: a second heat sink; 32: a second heating plate; 33: a second heat-conducting plate; 331: a second projection; 34: a second heat insulating frame; 341: a second through hole; 342: a guide groove; 35: a second fan;
4: a mounting bracket assembly; 41: a support frame; 42: an auxiliary guide unit; 421: a connecting frame; 422: a guide rail;
5: a lifting mechanism; 51: a first driving member; 52: a first lead screw; 53: a first slider;
6: an optical detection module; 61: an excitation light source; 62: a first lens; 63: a second lens; 64: a third lens; 65: a first optical filter; 66: a second optical filter; 67: a dichroic mirror; 68: a photodetector;
7: a horizontal movement mechanism; 71: a second driving member; 72: a second lead screw; 73: a second slider; 74: a push rod; 75: a supporting seat; 76: a third slider; 77: a guide rail; 78: a connecting seat;
8: a first mounting seat;
9: and a second mounting seat.
Detailed Description
In order to better understand the above technical solutions, exemplary embodiments of the present invention will be described in more detail below with reference to the accompanying drawings. While exemplary embodiments of the invention are shown in the drawings, it should be understood that the invention can be embodied in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.
Example one
As shown in fig. 1 to 3, the present invention provides an amplification apparatus for nucleic acid detection, which includes a microfluidic chip 1, a first temperature control module 2, a second temperature control module 3, a mounting rack assembly 4, a lifting mechanism 5, an optical detection module 6, and a horizontal movement mechanism 7.
The micro-fluidic chip 1 comprises a detection sheet which is of a rectangular structure, the micro-fluidic chip 1 with different shapes can be designed according to actual conditions, and a plurality of chambers are formed in the detection sheet.
The first side of the detection sheet is provided with a first sealing film for sealing the first side of the chamber, and the second side of the detection sheet is provided with a second sealing film for sealing the second side of the chamber. Wherein, the cavity is the through-hole, and the cavity is convenient for make the time and is convenient for first temperature control module 2 and second temperature control module 3 to go on heating up and cooling down fast to the both sides of sample in the cavity for through-hole and first seal membrane and the cooperation of second seal membrane.
The first temperature control module 2 in the amplification device for nucleic acid detection is in contact with the first sealing film and is used for heating the first side of the chamber in the microfluidic chip 1, and the second temperature control module 3 is in contact with the second sealing film and is used for heating the second side of the chamber in the microfluidic chip 1. As shown in fig. 1 to 3, the first temperature control module 2 and the second temperature control module 3 are arranged to face each other in the up-down direction.
As shown in fig. 1, the lifting mechanism 5 is disposed on the mounting frame assembly 4, and the lifting mechanism 5 is connected to the first temperature control module 2 to drive the first temperature control module 2 to move up and down, when the lifting mechanism 5 drives the first temperature control module 2 to move up and down, a floating space for placing the microfluidic chip 1 is reserved by increasing a distance between the first temperature control module 2 and the second temperature control module 3 so as to place the microfluidic chip 1, and after the microfluidic chip 1 is placed, the lifting mechanism 5 drives the first temperature control module 2 to move down to make the first temperature control module 2 contact with the first sealing film of the chamber. As shown in fig. 3, the lifting mechanism 5 includes a first driving member 51, a first lead screw 52, and a first sliding block 53 sleeved on the first lead screw 52, wherein the first driving member 51 drives the first lead screw 52 to rotate, so that the first sliding block 53 reciprocates up and down.
As shown in fig. 2, the mounting frame assembly 4 includes a supporting frame 41, a first driving member 51 of the lifting mechanism 5 is disposed at a top end of the supporting frame 41, the first temperature control module 2 is disposed in the first mounting seat 8, a first lead screw 52 on the lifting mechanism 5 penetrates through the supporting frame 41, a first slider 53 on the lifting mechanism 5 is fixedly connected with a top end of the first mounting seat 8, and when the first driving member 51 drives the first lead screw 52 to rotate, the first slider 53 moves along the first lead screw 52 and then drives the first temperature control module 2 to move up and down through the first mounting seat 8. In order to make the lifting of the first mounting seat 8 more stable, the mounting bracket assembly 4 further includes an auxiliary guide unit 42, one side of the auxiliary guide unit 42 is connected with the support frame 41, and the other side of the auxiliary guide unit 42 is connected with the first mounting seat 8.
Wherein, supplementary guide unit 42 is including setting up the linking frame 421 in the relative both sides of carriage 41, guided way 422 and the slider of setting on first mount pad 8 of setting on linking frame 421 (not shown), the slider passes through the connecting plate to be connected with first mount pad 8, slider and guided way 422 sliding connection, when first driving piece 51 drive first mount pad 8 drives the 2 elevating movement of first temperature control module on it, the slider slides along guided way 422, so that first mount pad 8 steady movement and then avoided rocking of 2 elevating movement in-process of first temperature control module.
In the present embodiment, the second temperature control module 3 is disposed on the mounting surface by the second mounting seat 9. The optical detection module 6 is used for generating a detection light beam to detect a sample, and the horizontal moving mechanism 7 is connected with the optical detection module 6 and drives the optical detection module 6 to horizontally move along the length direction of the microfluidic chip 1 so as to sequentially detect the sample in the heated chamber in the microfluidic chip 1. As shown in fig. 4, the optical detection module 6 includes an excitation light source 61, a first lens 62, a second lens 63, a third lens 64, a first filter 65, a second filter 66, a dichroic mirror 67, and a photodetector 68, the excitation light source 61 can select different wavelengths according to amplification detection requirements, and preferably, the excitation light source 61 uses LEDs to reduce instrument cost and volume of the optical detection module 6, wherein the excitation light source 61 is preferably a small-sized and low-energy-consumption LED, which can meet the requirement of a more compact structural design. In the practical application process, the lens uses the aspheric lens to further ensure the compactness of the whole structure of the optical detection module and the collection efficiency of the optical signal, which is also beneficial to the design requirement of small-volume instruments.
As shown in fig. 5, the optical detection module 6 works according to the following principle: light emitted by the excitation light source 61 is shaped by the first lens 62 and then filtered by the first optical filter 65 to obtain a light source with a specific wavelength, the light source is reflected by the dichroic mirror 67 to the third lens 64, focused and irradiated to the microfluidic chip to be detected, a sample to be detected in the microfluidic chip is excited to emit fluorescence, the fluorescence is shaped by the third lens 64, filtered by the dichroic mirror 67 to the second optical filter 66, and focused to the photoelectric detector 68 by the second lens 63, and then the detection of a fluorescence signal is completed.
As shown in fig. 2 to 3, the horizontal movement mechanism 7 includes a second driving member 71, a second lead screw 72, a second slider 73, a push rod 74, and a connecting seat 78. The second driving member 71 is disposed on the supporting seat 75, the second driving member 71 is connected to the second lead screw 72 and drives the second lead screw 72 to rotate, the second sliding block 73 is sleeved on the second lead screw 72, in this embodiment, the second sliding block 73 is respectively connected to the two connecting seats 78 through the two push rods 74, the optical detection module 6 is disposed on the connecting seats 78, and when the second driving member 71 drives the second lead screw 72 to rotate, the second sliding block 73 moves along the second lead screw 72 horizontally and linearly, and then pushes the connecting seats 78 to move horizontally through the push rods 74, so that the optical detection module 6 on the connecting seats 78 sequentially performs amplification detection on the chambers in the microfluidic chip 1. In order to avoid the shake of the optical detection module 6 during movement, the horizontal movement mechanism 7 further includes a third slider 76 and a guide rail 77, wherein the guide rail 77 is connected to the second mounting base 9 through a mounting frame, the extending direction of the guide rail 77 is the same as the moving direction of the second slider 73, the third slider 76 is disposed at the bottom end of the connecting base 78, and the third slider 76 is slidably connected to the guide rail 77.
As shown in fig. 6 and 7, the first temperature control module 2 includes a first heat sink 21, a first heating plate 22, a first heat-conducting plate 23 and a first heat-insulating frame 24 connected in sequence, and the first heat-conducting plate 23 partially extends out of the first heat-insulating frame 24 to contact the first sealing film and heat the first side of the chamber. The first heating plate 22 and the first heat conducting plate 23 can be insulated by arranging the first heat insulation frame 24 to avoid heat loss, and the rest parts of the microfluidic chip 1 except the cavity are prevented from being heated to avoid the damage of the microfluidic chip 1.
In practical applications, the first heating plate 22 may preferably be an electric heating film, an electric heating ceramic plate, or a semiconductor cooling plate, and the first heat conducting plate 23 is made of a heat conducting metal block. Among them, between the first heat sink 21 and the first heating plate 22, and between the first heating plate 22 and the first heat conduction plate 23, better heat conduction can be achieved through a gap heat conduction medium, and the gap heat conduction medium can use heat conduction silicone grease, phase change heat conduction material, or high-strength heat conduction film, etc.
As shown in fig. 8, a first accommodating groove for accommodating the first heating plate 22 and the first heat conducting plate 23 is concavely formed on a first side surface of the first heat insulating frame 24, at least one first through hole 241 is formed on a second side surface of the first heat insulating frame 24, and the first through hole 241 is communicated with the first accommodating groove. In order to improve the detection accuracy of the optical detection module 6, the light-shielding parts 242 extend from two sides of the first heat-insulating frame 24 to the second side surface of the first heat-insulating frame 24, the light-shielding parts 242 are provided with a plurality of detection grooves 2421 at intervals, and the detection grooves 2421 correspond to the side walls of the inner cavity of the microfluidic chip 1 one by one. Wherein, the first heat conducting plate 23 is provided with a first protruding part 231 matching with the first through hole 241, and the first protruding part 231 can protrude out of the first through hole 241 so as to directly contact the cavity in the microfluidic chip 1. In this embodiment, since the microfluidic chip 1 has two rows of cavities, in order to match the two rows of cavities, the first heat-insulating frame 24 is provided with two first through holes 241, and the first heat-conducting plate 23 is provided with two matching first protrusions 231. In practical applications, a stop 243 is further disposed on the second side of the first heat insulating frame 24 to allow the micro fluidic chip 1 to reach a precise mounting position.
As shown in fig. 9-10, the second temperature control module 3 includes a second heat sink 31, a second heating plate 32, a second heat-conducting plate 33 and a second heat-insulating frame 34 connected in sequence, the second heat-conducting plate 33 partially extends out of the second heat-insulating frame 34 to contact the second sealing film and heat the second side of the chamber. The second heat insulating frame 34 can keep the second heating plate 32 and the second heat transfer plate 33 warm to prevent heat loss.
As shown in fig. 11, a second accommodating groove for accommodating the second heating plate 32 and the second heat conducting plate 33 is concavely provided on a first side surface of the second heat insulating frame 34, at least one second through hole 341 is provided on a second side surface of the second heat insulating frame 34, the second through hole 341 is communicated with the second accommodating groove, in order to facilitate the installation of the microfluidic chip 1, a guide groove 342 is further provided on the second side surface of the second heat insulating frame 34 along the length direction thereof, wherein a cover sheet in the microfluidic chip 1 forms a guide block and extends into the guide groove 342, the second heat conducting plate 33 is provided with a second protrusion 331, the second protrusion 331 extends out of the second through hole 341, in this embodiment, since the microfluidic chip 1 is provided with two rows of chambers, the second heat insulating frame 34 is provided with two second through holes 341, and the second heat conducting plate 33 is provided with two second protrusions 331.
In this embodiment, in order to improve the heat dissipation effect in the first temperature control module 2 and the second temperature control module 3, the first temperature control module 2 further includes a first fan 25, the first fan 25 is located at one side of the fins in the first heat sink 21 and connected to the first mounting seat 8, the second temperature control module 3 further includes a second fan 35, and the second fan 35 is located at one end of the fins in the second heat sink 31 and connected to the second mounting seat 9.
The amplification device for nucleic acid detection provided by the embodiment is provided with the first temperature control module 2 and the second temperature control module 3 which are matched with the first sealing film and the second sealing film in the microfluidic chip 1, so that the first side and the second side of the cavity in the microfluidic chip 1 can be subjected to rapid temperature change treatment simultaneously, the sample can be detected within 0.5-1 hour, the PCR reaction time can be greatly shortened, the nucleic acid detection time is shortened, and due to the temperature change of the two sides of the cavity, when the temperature reaches a stable state, the heat loss of the cavity to the outside is less, the uniformity of heating the sample in the cavity is good, and the amplification detection efficiency is further improved. Meanwhile, multiple chambers in the microfluidic chip 1 can realize PCR detection of more targets and have a simple structure.
Example two
The embodiment provides a detection method based on an amplification device, which adopts the amplification device for nucleic acid detection in the first embodiment, the amplification device further comprises a controller, the controller is respectively connected with the first temperature control module 2, the second temperature control module 3 and the optical detection module 6 in a communication way, and the detection method comprises the following steps:
s1, a lifting mechanism 5 drives a first temperature control module 2 to ascend so as to reserve a floating space for placing a micro-fluidic chip 1, and the micro-fluidic chip 1 is placed conveniently;
s2, placing the microfluidic chip 1 on the second temperature control module 3, driving the first temperature control module 2 to descend by the lifting mechanism 5 so as to press the microfluidic chip 1 tightly, and enabling the first temperature control module 2 and the second temperature control module 3 to be in contact with a first sealing film and a second sealing film of the microfluidic chip 1 respectively;
s3, the controller controls the first temperature control module 2 and the second temperature control module 3 to respectively heat the first side and the second side of the cavity in the microfluidic chip 1;
and S4, the controller controls the horizontal moving mechanism 7 to drive the optical detection module 6 to horizontally move along the length direction of the microfluidic chip 1 so as to sequentially detect the samples in the heated chambers in the microfluidic chip 1.
The detection method of the amplification device provided by the embodiment can simultaneously carry out temperature-changing treatment on the samples in the chambers on the first side and the second side in the chamber of the microfluidic chip 1, greatly shorten the PCR reaction time, further shorten the nucleic acid detection time, and can realize PCR detection of more targets by matching with a plurality of chambers in the microfluidic chip 1.
In the description of the present invention, it is to be understood that the terms "first", "second" and the like are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implying any number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless specifically defined otherwise.
In the present invention, unless otherwise expressly stated or limited, the terms "mounted," "connected," "secured," and the like are to be construed broadly and can, for example, be fixedly connected, detachably connected, or integrally formed; can be mechanically or electrically connected; can be directly connected or indirectly connected through an intermediate medium; either as communication within the two elements or as an interactive relationship of the 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.
In the description herein, the description of the terms "one embodiment," "some embodiments," "an embodiment," "an example," "a specific example" or "some examples" or the like, means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above are not necessarily intended to refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, various embodiments or examples and features of different embodiments or examples described in this specification can be combined and combined by one skilled in the art without contradiction.
Although embodiments of the present invention have been shown and described above, it should be understood that the above embodiments are illustrative and not restrictive, and that those skilled in the art may make changes, modifications, substitutions and alterations to the above embodiments without departing from the scope of the present invention.
Claims (7)
1. An amplification device for nucleic acid detection, comprising:
the micro-fluidic chip (1) comprises a detection sheet, wherein the detection sheet is of a rectangular structure and is provided with a plurality of communicated chambers, a first sealing film is arranged on the first side of the micro-fluidic chip (1) to seal the first side of the chambers, and a second sealing film is arranged on the second side of the micro-fluidic chip (1) to seal the second side of the chambers;
a first temperature control module (2) in contact with the first sealing membrane and adapted to heat a first side of the chamber;
the second temperature control module (3) is in contact with the second sealing film and used for heating the second side of the chamber, and the first temperature control module (2) and the second temperature control module (3) are arranged oppositely from top to bottom;
a mounting bracket assembly (4);
the lifting mechanism (5) is arranged on the mounting frame assembly (4) and is connected with the first temperature control module (2) to drive the first temperature control module (2) to move up and down;
an optical detection module (6) for generating a detection beam for detecting the sample;
the horizontal moving mechanism (7) is connected with the optical detection module (6) and drives the optical detection module (6) to horizontally move along the length direction of the microfluidic chip (1) so as to sequentially detect the samples in the heated chambers in the microfluidic chip (1);
the first temperature control module (2) comprises a first radiator (21), a first heating plate (22), a first heat conducting plate (23) and a first heat insulation frame (24) which are sequentially connected;
the first heat-conducting plate (23) partially protrudes beyond the first heat-insulating frame (24) to contact the first sealing film and heat the first side of the chamber;
a first side surface of the first heat insulation frame (24) is concavely provided with a first placing groove for accommodating the first heating plate (22) and the first heat conduction plate (23);
the second side surface of the first heat insulation frame (24) is provided with at least one first through hole (241), the first through hole (241) is communicated with the first containing groove, the first heat conduction plate (23) is provided with a first protruding part (231), and the first protruding part (231) extends out of the first through hole (241);
two sides of the first heat insulation frame (24) extend to the second side face of the first heat insulation frame (24) to form a light shielding part (242), the light shielding part (242) is provided with a plurality of detection grooves (2421) at intervals, and the detection grooves (2421) correspond to the side walls of the chambers in the microfluidic chip (1) one by one;
the second temperature control module (3) comprises a second radiator (31), a second heating plate (32), a second heat conducting plate (33) and a second heat insulation frame (34) which are connected in sequence;
the second heat-conducting plate (33) partially protrudes beyond the second heat-insulating frame (34) to contact the second sealing film and heat the second side of the chamber.
2. The amplification apparatus for nucleic acid detection according to claim 1, wherein:
a second placing groove for accommodating the second heating plate (32) and the second heat conducting plate (33) is concavely arranged on the first side surface of the second heat insulation frame (34);
the second side surface of the second heat insulation frame (34) is provided with at least one second through hole (341), the second through hole (341) is communicated with the second placing groove, the second heat conduction plate (33) is provided with a second protruding part (331), and the second protruding part (331) extends out of the second through hole (341);
the second side surface of the second heat insulation frame (34) is also provided with a guide groove (342) along the length direction.
3. The amplification apparatus for nucleic acid detection according to claim 1, wherein:
the mounting frame assembly (4) comprises a support frame (41);
the lifting mechanism (5) is arranged at the top end of the supporting frame (41), the first temperature control module (2) is arranged in the first mounting seat (8), and the lifting mechanism (5) is connected with the first temperature control module (2) through the first mounting seat (8);
the second temperature control module (3) is arranged on the mounting surface through a second mounting seat (9);
the mounting frame assembly (4) further comprises an auxiliary guide unit (42), one side of the auxiliary guide unit (42) is connected with the support frame (41), and the other side of the auxiliary guide unit (42) is connected with the first mounting seat (8).
4. The amplification apparatus for nucleic acid detection according to claim 3, wherein:
the first temperature control module (2) further comprises a first fan (25), and the first fan (25) is located on one side of the first radiator (21) and connected with the first mounting seat (8);
the second temperature control module (3) further comprises a second fan (35), and the second fan (35) is located at one end of the second radiator (31) and connected with the second mounting seat (9).
5. The amplification apparatus for nucleic acid detection according to claim 2, wherein:
a stop part (243) is arranged on the second side surface of the first heat insulation frame (24).
6. The amplification apparatus for nucleic acid detection according to claim 3, wherein:
the auxiliary guide unit (42) comprises a connecting frame (421) arranged on two opposite sides of the supporting frame (41), a guide rail (422) arranged on the connecting frame (421) and a sliding block arranged on the first mounting seat (8), and the sliding block is connected with the guide rail (422) in a sliding mode.
7. An amplification apparatus-based detection method, wherein the amplification apparatus for nucleic acid detection according to any one of claims 1 to 6 is used, the amplification apparatus further comprises a controller, the controller is respectively connected to the first temperature control module (2), the second temperature control module (3) and the optical detection module (6) in communication, and the detection method comprises the following steps:
s1, a lifting mechanism (5) drives a first temperature control module (2) to rise so as to reserve a floating space for placing a micro-fluidic chip (1);
s2, placing the microfluidic chip (1) on a second temperature control module (3), driving the first temperature control module (2) to descend by the lifting mechanism (5) to compress the microfluidic chip (1), and enabling the first temperature control module (2) and the second temperature control module (3) to be in contact with a first sealing film and a second sealing film of the microfluidic chip (1) respectively;
s3, the controller controls the first temperature control module (2) and the second temperature control module (3) to respectively heat the first side and the second side of the cavity in the microfluidic chip (1);
and S4, the controller controls the horizontal moving mechanism (7) to drive the optical detection module (6) to horizontally move along the length direction of the microfluidic chip (1) so as to sequentially detect the samples in the heated chambers in the microfluidic chip (1).
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