CN114177963A - Nucleic acid analysis device - Google Patents
Nucleic acid analysis device Download PDFInfo
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- CN114177963A CN114177963A CN202210036525.5A CN202210036525A CN114177963A CN 114177963 A CN114177963 A CN 114177963A CN 202210036525 A CN202210036525 A CN 202210036525A CN 114177963 A CN114177963 A CN 114177963A
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- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 48
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 44
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 44
- 238000004458 analytical method Methods 0.000 title claims abstract description 26
- 230000003321 amplification Effects 0.000 claims abstract description 54
- 238000003199 nucleic acid amplification method Methods 0.000 claims abstract description 54
- 238000000605 extraction Methods 0.000 claims abstract description 53
- 230000007246 mechanism Effects 0.000 claims abstract description 50
- 238000001514 detection method Methods 0.000 claims abstract description 26
- 230000003287 optical effect Effects 0.000 claims abstract description 7
- 230000001105 regulatory effect Effects 0.000 claims abstract description 4
- 238000007789 sealing Methods 0.000 claims abstract description 3
- 238000010438 heat treatment Methods 0.000 claims description 26
- 238000009413 insulation Methods 0.000 claims description 12
- 230000005540 biological transmission Effects 0.000 claims description 8
- 230000001276 controlling effect Effects 0.000 claims description 6
- 238000000034 method Methods 0.000 abstract description 8
- 230000008569 process Effects 0.000 abstract description 8
- 238000012408 PCR amplification Methods 0.000 description 25
- 230000005284 excitation Effects 0.000 description 11
- 239000004020 conductor Substances 0.000 description 5
- 238000010586 diagram Methods 0.000 description 5
- 239000000835 fiber Substances 0.000 description 5
- 238000001917 fluorescence detection Methods 0.000 description 5
- 239000013307 optical fiber Substances 0.000 description 5
- 239000007788 liquid Substances 0.000 description 4
- 238000003752 polymerase chain reaction Methods 0.000 description 4
- 238000002203 pretreatment Methods 0.000 description 4
- 239000002470 thermal conductor Substances 0.000 description 4
- 238000013459 approach Methods 0.000 description 3
- 238000005192 partition Methods 0.000 description 3
- 238000010828 elution Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000004065 semiconductor Substances 0.000 description 2
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000001506 fluorescence spectroscopy Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000007781 pre-processing Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 125000006850 spacer group Chemical group 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
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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
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
-
- 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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- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Clinical Laboratory Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Hematology (AREA)
- Dispersion Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Automatic Analysis And Handling Materials Therefor (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The present invention relates to a nucleic acid analysis device including: the centrifugal driving mechanism is used for driving the micro-fluidic chip to rotate and comprises a rotating shaft, and the rotating shaft is used for being connected with the micro-fluidic chip; the extraction temperature control mechanism comprises a top temperature control unit and a bottom temperature control unit which are positioned on two sides of the microfluidic chip; the amplification temperature control mechanism is positioned beside the extraction temperature control mechanism and comprises an upper temperature control module and a lower temperature control module, and the upper temperature control module and the lower temperature control module are used for sealing the amplification cavity and regulating the temperature of the amplification cavity when the amplification cavity carries out amplification reaction; and the detection mechanism forms an optical detection channel with the amplification cavity through the amplification temperature control module. The nucleic acid analysis device controls the temperature of different extraction temperature areas on the microfluidic chip and independently controls the temperature of the amplification cavity, so that the temperature influence among different areas of the microfluidic chip is reduced, and the detection process is convenient and quick.
Description
Technical Field
The present invention relates to the field of biomedical technology, and in particular, to a nucleic acid analyzer.
Background
Nucleic acid detection is one type of molecular diagnostics. It is widely used because of its high accuracy and early diagnostic timeliness. The detection process can be divided into: sampling, extracting nucleic acid, amplifying, detecting and the like. The nucleic acid extraction process generally requires the sequential steps of lysis, washing and elution to remove substances present in the sample that may have an effect on the amplification step. Each step needs to add different reagents correspondingly and is carried out at a certain temperature, so that pure nucleic acid is finally obtained for subsequent amplification.
Nucleic acid amplification usually employs Polymerase Chain Reaction (PCR) to amplify a nucleic acid sequence to be detected, and the purpose of amplification is to amplify a target nucleic acid, which is a trace and difficult to detect, to a large amount and then to easily detect it. PCR amplification is generally performed for 40 cycles, and one cycle can be divided into three steps of denaturation, annealing and extension, and theoretically, the number of target nucleic acid fragments is doubled after one cycle. Wherein, each step is realized by the temperature change of the PCR reaction system, namely, each step corresponds to one temperature. It is known that the key to the realization of PCR amplification lies in the rapid and accurate temperature control. In the PCR amplification reaction system, a target nucleic acid fragment is marked by utilizing a fluorescent group, and the fluorescence becomes stronger along with the amplification of the target nucleic acid fragment. The fluorescence data is collected in real time through the optical module, so that an S-shaped amplification curve can be drawn for analysis, and detection is further completed.
Because the nucleic acid detection process has more steps, in the existing scheme for completing the whole nucleic acid detection, a PCR instrument and a nucleic acid extraction instrument are used, and auxiliary equipment such as a blending instrument, a centrifugal machine, a liquid transfer gun and the like is also needed, so that the sample circulation is complicated, the detection process is complex, and the cost is higher.
Disclosure of Invention
In view of this, it is necessary to provide a nucleic acid analyzer in order to solve the problem of redundancy of the conventional nucleic acid detecting apparatus.
A nucleic acid analysis device for analyzing a sample in a microfluidic chip, the microfluidic chip comprising an extraction region and an amplification chamber, the extraction region comprising a plurality of extraction temperature zones, comprising:
the centrifugal driving mechanism is used for driving the micro-fluidic chip to rotate and comprises a rotating shaft, and the rotating shaft is used for being connected with the micro-fluidic chip;
the extraction temperature control mechanism comprises a top temperature control unit and a bottom temperature control unit which are positioned at two sides of the microfluidic chip and are used for respectively controlling the air temperature of different extraction temperature areas on the microfluidic chip;
the amplification temperature control mechanism is positioned beside the extraction temperature control mechanism and comprises an upper temperature control module and a lower temperature control module, and the upper temperature control module and the lower temperature control module are used for sealing the amplification cavity and regulating the temperature of the amplification cavity when the amplification cavity carries out amplification reaction;
and the detection mechanism forms an optical detection channel with the amplification cavity through the amplification temperature control module.
In one embodiment, the top temperature control unit comprises a top heat insulation cover and a top heating element, the top heating element is located in the top heat insulation cover, the bottom temperature control unit comprises a bottom heat insulation cover and a bottom heating element, the bottom heating element is located in the bottom heat insulation cover, and when the microfluidic chip is connected to the rotating shaft, the top heat insulation cover and the bottom heat insulation cover are both not in contact with the microfluidic chip.
In one embodiment, the top temperature control unit further includes a top fan structure and a top deflector disc located in the top heat insulation cover, and the top fan structure is sleeved on the rotating shaft and located on one side of the top deflector disc; the bottom temperature control unit further comprises a bottom fan blade structure, a bottom flow guide disc and an idle shaft, wherein the bottom fan blade structure, the bottom flow guide disc and the idle shaft are located in the bottom heat insulation cover, the bottom fan blade structure is sleeved on the idle shaft and located on one side of the bottom flow guide disc, and the idle shaft is used for being connected with the rotating shaft to rotate synchronously along with the rotating shaft.
In one embodiment, the top deflector disc and the bottom deflector disc are both provided with a through air inlet and at least one air outlet.
In one embodiment, a through air inlet is arranged in the central area of the top flow guide disc, and a through air outlet is arranged at the edge of the top flow guide disc; the central area of the bottom flow guide disc is provided with a through central air inlet, and the bottom flow guide disc is provided with an inner ring air outlet and an outer ring air outlet along the radial direction.
In one embodiment, the bottom deflector comprises a first air distribution plate and a second air distribution plate which are stacked, and the first air distribution plate and the second air distribution plate can rotate relatively to enable one of the inner ring air outlet and the outer ring air outlet to be communicated with the central air inlet.
In one embodiment, a containing groove is disposed on the upper temperature control module or the lower temperature control module, and the containing groove is used for containing the amplification cavity.
In one embodiment, the amplification temperature control mechanism further comprises a driving module, the movable module comprises an upper connecting rod, a lower connecting rod, and a vertical sliding block guide rail and a horizontal sliding block guide rail, the vertical sliding block guide rail comprises an upper sliding block connected with the upper temperature control module and a lower sliding block connected with the lower temperature control module, one end of the upper connecting rod is hinged to the upper sliding block, the other end of the upper connecting rod is hinged to the sliding block on the horizontal sliding block guide rail, one end of the lower connecting rod is hinged to the lower sliding block, and the other end of the lower connecting rod is hinged to the sliding block on the horizontal sliding block guide rail.
In one embodiment, the driving module further includes a transmission shaft and a cam sleeved on the transmission shaft, the cam abuts against the upper temperature control module, and the upper temperature control module and the lower temperature control module can approach or separate from each other in the rotation process of the cam.
In one embodiment, the amplification temperature control mechanism further includes an elastic member, one end of the elastic member is connected to the upper temperature control module, the other end of the elastic member is connected to the lower temperature control module, when the upper temperature control module and the lower temperature control module are away from each other, the elastic member is in a deformed state, and when the upper temperature control module and the lower temperature control module are in contact with each other, the elastic member is in an original state.
On one hand, the nucleic acid analysis device adopts the microfluidic chip to integrate nucleic acid extraction, amplification and analysis, thereby reducing equipment investment. On the other hand, the extraction temperature control mechanism controls the temperature of different extraction temperature areas on the micro-fluidic chip, so that the temperature influence among the extraction temperature areas can be reduced, and the amplification temperature control mechanism controls the temperature of an amplification cavity on the micro-fluidic chip independently, so that the temperature required by the amplification cavity can be controlled accurately and quickly. On the other hand, an optical detection channel can be formed between the detection mechanism and the amplification cavity, and the detection process is convenient and quick.
Drawings
FIG. 1 is a schematic structural view of a nucleic acid analysis apparatus according to an embodiment of the present invention.
Fig. 2 is a schematic structural diagram of a microfluidic chip according to an embodiment of the present invention.
FIG. 3 is a schematic structural diagram of a centrifugal temperature control device in the nucleic acid analysis apparatus according to an embodiment of the present invention.
Fig. 4 is a cross-sectional view of fig. 3.
FIG. 5 is a cross-sectional view of a top fan blade structure and a top deflector in a nucleic acid analysis device according to an embodiment of the present invention.
FIG. 6 is a schematic structural diagram of a bottom baffle in the nucleic acid analysis apparatus according to an embodiment of the present invention.
FIG. 7 is a schematic diagram of an amplification temperature control mechanism of a nucleic acid analysis apparatus according to an embodiment of the present invention.
FIG. 8 is a schematic structural view of an amplification temperature control mechanism of a nucleic acid analysis apparatus according to an embodiment of the present invention, with a support frame omitted.
FIG. 9 is a schematic view of the nucleic acid analyzer from another perspective in one embodiment of the present invention.
Detailed Description
This invention can be embodied in many different forms than those herein described and many modifications may be made by those skilled in the art without departing from the spirit of the invention.
In the description of the present invention, the terms "vertical", "horizontal", "upper", "lower", "left", "right", "center", "longitudinal", "lateral", "length", and the like are used to indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, only for the convenience of description of the present invention and for simplicity of description. The first feature may be directly on or directly under the second feature or may be indirectly on or directly under the second feature via intervening media. The terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. In the description of the present invention, "a plurality" means at least two, e.g., two, three, etc., unless specifically limited otherwise.
In the present invention, the terms "mounted," "connected," "secured," and the like are to be construed broadly unless otherwise specifically indicated and limited. When an element is referred to as being "secured to" or "disposed on" another element, it can be directly on the other element or intervening elements may also be present. When an element is referred to as being "connected" to another element, it can be directly connected to the other element or intervening elements may also be present.
Referring to fig. 1, fig. 1 is a schematic structural diagram of a nucleic acid analysis apparatus according to an embodiment of the present invention, and the nucleic acid analysis apparatus according to the embodiment of the present invention may be used in conjunction with a microfluidic chip 10 to implement molecular diagnosis, such as nucleic acid detection. The micro-fluidic chip 10 integrates nucleic acid extraction and PCR amplification functions, and before the PCR amplification function is performed, pretreatment such as cracking-cleaning and elution is required to be performed on a sample to realize nucleic acid extraction and purification. These pre-treatment sections need to be performed at different temperatures, and the temperatures required for the pre-treatment sections often differ from the temperatures required for PCR amplification. For this purpose, the nucleic acid analysis apparatus includes a centrifugal temperature control device 20, an amplification temperature control mechanism 300, and a detection mechanism 500, wherein the centrifugal temperature control device 20 is used for providing a liquid flow driving force for the microfluidic chip 10 and controlling the temperature during the pre-processing. The amplification temperature control mechanism 300 is used for temperature control of PCR amplification. The detection mechanism 500 is used for detecting the product in the amplification chamber on the microfluidic chip 10 after the microfluidic chip 10 completes the PCR amplification.
Referring to fig. 2, in the present embodiment, the microfluidic chip 10 is a centrifugal microfluidic chip, and is in a disc shape, and is provided with an extraction region for performing a pre-treatment and a PCR amplification chamber, because the temperatures required in the steps of the extraction region are different, in the present embodiment, the part of the microfluidic chip 10 located in the extraction region is named as a first extraction temperature region a, a second extraction temperature region B, and a third extraction temperature region C according to different functions to be implemented, and the first extraction temperature region, the second extraction temperature region, and the third extraction temperature region are concentric annular regions. The PCR amplification chamber is marked with P in the figure and is positioned at the edge of the microfluidic chip 10. The PCR amplification cavity protrudes out of the surface of the microfluidic chip 10. The center of the microfluidic chip 10 is provided with a spline hole for connecting with the centrifugal temperature control device 20.
Referring to FIG. 3, FIG. 3 is a schematic structural view of a centrifugal temperature control device 20 in a nucleic acid analysis apparatus according to an embodiment of the present invention. The centrifugal temperature control device 20 includes a frame 21, and a bracket 23, an extraction temperature control mechanism 100, a centrifugal drive mechanism 900, and a lifting mechanism 200, which are located on the frame 21. The extraction temperature control mechanism 100 includes a top temperature control unit 110 and a bottom temperature control unit 130, which are vertically spaced. The top temperature control unit 110 is connected to the frame 21 and the bottom temperature control unit 130 is connected to the lift mechanism 200. The bracket 23 is used for pushing the microfluidic chip 10 to a set position, the lifting mechanism 200 is used for pushing the microfluidic chip 10 to be connected with the centrifugal driving mechanism 900, and the top temperature control unit 110 and the bottom temperature control unit 130 are respectively located at two sides of the microfluidic chip 10 and used for regulating and controlling the temperature of different areas of the microfluidic chip 10. When the microfluidic chip 10 is mounted on the centrifugal driving mechanism 900, the top temperature control unit 110 and the bottom temperature control unit 130 on both sides of the microfluidic chip 10 are not in contact with the microfluidic chip 10.
In this embodiment, the top temperature control unit 110 is used for controlling the air temperature above the first extraction temperature zone of the microfluidic chip 10, and the bottom temperature control unit 130 is used for controlling the air temperature above the second extraction temperature zone and the air temperature above the third extraction temperature zone of the microfluidic chip 10.
Referring to fig. 3, the centrifugal driving mechanism 900 includes a driving motor 920 and a rotating shaft 930, the driving motor 920 is located at the top end of the frame 21, a power output end of the driving motor 920 is connected to one end of the rotating shaft 930, and the other end of the rotating shaft 930 is used for being connected to the microfluidic chip 10. In one embodiment, the end of the rotating shaft 930 for connecting to the microfluidic chip 10 is provided with a chuck, so as to connect to the spline hole of the microfluidic chip 10 through the chuck.
Referring to fig. 4, the top temperature control unit 110 includes a top heat insulating cover 111 and a top heating element 120. The top thermal shield 111 is fixed to the frame 21 on a side facing away from the driving motor 920. The top heat insulating cover 111 is hollow, the top heating element 120 is located in the top heat insulating cover 111, and the rotating shaft 930 is partially inserted into the top heat insulating cover 111. The top heating member 120 is used to generate heat and conduct the heat out to heat the air in the area covered by the heat insulating cover 111, so as to heat the corresponding area on the microfluidic chip 10 connected to the rotating shaft 930. The side of the top thermal shield 111 away from the top of the frame 21 has a small gap with the microfluidic chip 10.
In this embodiment, the top heating element 120 is a spiral-shaped annular resistance wire whose radial dimension is adapted to the first temperature range.
Referring to fig. 4, the top temperature control unit 110 further comprises a top airflow circulation assembly located within the top thermal shield 111, the top airflow circulation assembly comprising a top fan blade structure 113 and a top deflector 115. The top baffle 115 is connected to an inner wall of the top heat insulating cover 111, and divides an inner space of the top heat insulating cover 111 into an upper chamber and a lower chamber. The top heating element 120 and the top fan blade structure 113 are both located on the side of the top deflector 115 facing away from the microfluidic chip 10, i.e. both located in the upper chamber of the top thermal shield 111. The top heating member 120 surrounds the outside of the top fan blade structure 113. The top fan blade structure 113 is sleeved on the rotating shaft 930 and can rotate synchronously under the rotation of the rotating shaft 930.
For ease of viewing, fig. 5 shows a schematic cross-sectional view of top vane structure 113 and top diaphragm 115 in isolation. The top blade structure 113 includes a plurality of circumferentially disposed blades. The top deflector 115 is provided with an air inlet 112 and an air outlet 114 which are through, the air inlet 112 is formed in the central area of the top deflector 115, and the air outlet 114 is formed at the edge of the top deflector 115. The air outlets 114 are provided in a plurality and are uniformly distributed around the central axis of the top baffle 115. The upper chamber and the lower chamber inside the top thermal insulation cover 111 are communicated with the air outlet 114 through the air inlet 112. In this embodiment, the top heating element 120 is located at the air outlet 114 of the top deflector 115.
Referring to fig. 4 and 5, when the top fan blade structure 113 is driven by the rotating shaft 930 to rotate, a negative pressure is generated in the central region of the top fan blade structure 113, so as to cause the air in the top thermal insulation cover 111 to flow. Air flows into the intake 112 and out of the outtake 114 to create a flowing air stream, and so on. The air flow of the circulation flow drives the air flow in the vicinity of the top heating element 120, whereby the temperature of the air in the top heat insulating cover 111 can be made uniform. And because the air outlet 114 is located at the periphery of the air inlet 112, that is, the air inlet 112 is located in the area surrounded by the air outlet 114, and a small gap exists between the top heat insulating cover 111 and the microfluidic chip 10, an "air short circuit" is formed between the air outlet 114 and the air inlet 112, that is, flowing gas is constrained to flow tracks between the air outlet 114 and the air inlet 112 and cannot diffuse, thereby ensuring that the gas flows circularly inside the top heat insulating cover 111.
Referring to fig. 4, the bottom temperature control unit 130 includes a bottom heat insulating cover 131 and a bottom heating element 140. The bottom heat insulating cover 131 is hollow, and the bottom heating element 140 is located in the bottom heat insulating cover 131. There is a small gap in the vertical direction between the bottom thermal shield 131 and the microfluidic chip 10. The bottom heating member 140 is used for heating the air in the region covered by the bottom thermal insulation cover 131, so as to heat the corresponding region of the microfluidic chip 10.
The bottom heating element 140 is a helical, circular resistance wire.
Referring to fig. 4, similar to the top temperature control unit 110, the bottom temperature control unit 130 further comprises a bottom airflow circulation assembly located within the bottom thermal shield 131, the bottom airflow circulation assembly comprising a bottom fan blade structure 133, a bottom baffle 135, and an idler shaft 230. The bottom blade structure 133 and the top blade structure 113 have similar structures and different sizes, and the central axes of the two structures are located on the same straight line. The bottom diaphragm 135 is similar in structure and function to the top diaphragm 115. The axis of the idle shaft 230 is aligned with the axis of the rotation shaft 930. The bottom fan blade structure 133 and the bottom heating element 140 are both located on a side of the bottom deflector 135 facing away from the microfluidic chip 10. The bottom heating member 140 surrounds the bottom blade structure 133, and the bottom blade structure 133 is sleeved on the idle shaft 230. When the microfluidic chip 10 is connected to the rotating shaft 930, the idle shaft 230 is also connected to the rotating shaft 930, so that only one driving motor 920 is needed to drive the idle shaft 230 and the rotating shaft 930 to rotate simultaneously, and further drive the bottom blade structure 133 and the top blade structure 113 to rotate. In one embodiment, the idler shaft 230 is also coupled to the spindle 930 via the chuck.
When the bottom blade structure 133 rotates, the bottom deflector 135 and the bottom blade structure 133 cooperate with each other to form a circulating airflow in the bottom heat-insulating cover 131, so that heat generated by the bottom heating element 140 is uniformly distributed in the bottom heat-insulating cover 131.
When the bottom temperature control unit 130 needs to control at least two extraction temperature zones, for example, the second extraction temperature zone and the third extraction temperature zone of the microfluidic chip 10, it is required that the air with the set temperature in the bottom heat-insulating cover 131 can be used to heat the different extraction temperature zones in a targeted manner. In addition to the same points as above, the bottom baffle 135 and the top baffle 115 are different in that in the present embodiment, the bottom baffle 135 has a central air inlet, an inner ring air outlet and an outer ring air outlet, which are through, that is, a plurality of air outlets are radially arranged compared to the top baffle 115. The central air inlet is disposed in the central region of the bottom diaphragm 135, and the inner ring air outlet is closer to the center of the bottom diaphragm 135 than the outer ring air outlet. The airflow inlet is normally open, and the inner ring air outlet and the outer ring air outlet can be selectively opened or closed. When the bottom fan blade structure 133 rotates, the inner ring air outlet and the outer ring air outlet are respectively controlled to open and close, so that two different airflow circulation paths are provided in the bottom temperature control unit 130.
Referring to fig. 6, in one embodiment, the bottom baffle 135 includes first and second air distribution plates 137, 139 that are the same size. The first air distribution plate 137 is provided with a first air inlet 132, a first inner ring air outlet 134 and a first outer ring air outlet 136 which are communicated. The first air inlet 132 is formed in a central region of the first air distribution plate 137, and the first inner ring air outlet 134 is closer to a central point of the first air distribution plate 137 than the first outer ring air outlet 136. The first inner ring air outlet 134 and the first outer ring air outlet 136 are arranged in a plurality of numbers and are uniformly arranged in the circumferential direction. The second air distribution plate 139 is provided with a second air inlet 142, a second inner ring air outlet 144 and a second outer ring air outlet 146 which are through. The second air inlet 142 is formed in a central region of the second air distribution plate 139, and the second inner ring air outlet 144 is closer to a central point of the second air distribution plate 139 than the second outer ring air outlet 146. The second inner ring air outlet 144 and the second outer ring air outlet 146 are both provided in a plurality of numbers and are uniformly arranged in the circumferential direction.
In this embodiment, a connection line between the first inner ring outlet 134 and the center point of the first air distribution plate 137 and a connection line between the first outer ring outlet 136 and the center point of the first air distribution plate 137 are not located on the same radial line. The connecting line of the second inner ring air outlet 144 and the center point of the second air distribution plate 139 and the connecting line of the second outer ring air outlet 146 and the second air distribution plate 139 are positioned on the same radial line.
The first air distributor plate 137 and the second air distributor plate 139 are stacked to form the bottom deflector 135. The first intake vent 132 and the second intake vent 142 collectively form a central intake vent of the bottom diaphragm 135. When the first air distribution plate 137 and the second air distribution plate 139 rotate relatively, the first inner ring air outlet 134 and the second inner ring air outlet 144 are communicated, meanwhile, no air flow passes through the first outer ring air outlet 136 and the second outer ring air outlet 146, and the communicated first inner ring air outlet 134 and the second inner ring air outlet 144 form an inner ring air outlet of the bottom deflector 135 together; or the first outer ring air outlet 136 is communicated with the second outer ring air outlet 146, and meanwhile, no air flow passes through the first inner ring air outlet 134 and the second inner ring air outlet 144, at this time, the communicated first outer ring air outlet 136 and the second outer ring air outlet 146 together form an outer ring air outlet of the bottom deflector 135. By relatively rotating the first air distribution plate 137 and the second air distribution plate 139 to a predetermined angle, the central air inlet of the bottom deflector 135 can be respectively communicated with the inner ring air outlet or the outer ring air outlet.
In cooperation with different air outlets of the bottom flow guiding disc 135, the bottom temperature control unit 130 further includes a partition plate 138 located on the bottom flow guiding disc 135 and located on a side of the bottom flow guiding disc 135 facing the microfluidic chip 10. The spacer 138 has a small gap with the microfluidic chip 10. At least two partition plates 138 are respectively positioned between the central air inlet and the inner ring air outlet, between the inner ring air outlet and the outer ring air outlet, and between the outer ring air outlet and the side surface of the bottom deflector 135, so as to enclose different annular spaces to correspond to different extraction temperature regions on the microfluidic chip 10. The provision of different partition plates 138 avoids the influence of the air flow having a predetermined temperature on the adjacent extraction temperature zones.
It should be noted that, if the second extraction temperature region of the microfluidic chip 10 is subjected to the corresponding pre-treatment, the temperature is not affected any more, that is, even if the temperature of the second extraction temperature region is increased again during the heating process of the third extraction temperature region, the final nucleic acid extraction result is not affected.
Referring to fig. 3, the lifting mechanism 200 includes a lifting motor 250, a lift pin 260, and an idler plate 210. The bottom temperature control unit 130 is located above the idler plate 210, and the lift pin 260 is located below the idler plate 210. A slide rail structure is arranged between the left side and the right side of the idle disk 210 and the frame 21, the top rod 260 can push the idle disk 210 to move in the vertical direction relative to the frame 21 under the driving of the lifting motor 250, and the idle shaft 230 in the bottom temperature control unit 130 can push the microfluidic chip 10 in the bracket 23 to be connected with the rotating shaft 930.
Referring to fig. 4, the lifting mechanism 200 further includes an idle shaft support 220 and an elastic assembly 240, wherein the elastic assembly 240 includes a sheath 241 and an elastic element 243, and the elastic element 243 is located in the sheath 241. One end of the jacket 241 is connected with the top rod 260, and the other end is slidably sleeved on the idle shaft support 220. The idle shaft support 220 has one end abutting against the elastic member 243 located in the jacket 241 and the other end embedded in the center of the idle disk 210 to be rotatably fitted with the idle shaft 230. The elastic member 243 may buffer the sleeve 241 and the idle shaft support 220 when they are relatively moved.
After the bracket 23 sends the microfluidic chip 10 to a set position, the lifting motor 250 drives the sleeve 241 connected to the top rod 260 to move upward, the idle shaft support 220 moves upward under the support of the elastic element 243 in the sleeve 241, and further pushes the idle disk 210 to move upward, and the bottom temperature control unit 130 moves upward along with the idle disk 210. The idle shaft 230 is engaged with the microfluidic chip 10 during the upward movement, and pushes the microfluidic chip 10 to be disengaged from the bracket 23 until the microfluidic chip 10 is engaged with the rotating shaft 930. At this time, the rotation shaft 930, the microfluidic chip 10, and the idle shaft 230 are sequentially connected together. When the driving motor 920 drives the rotating shaft 930 to rotate, the microfluidic chip 10 and the idle shaft 230 also rotate synchronously, and further the top blade structure 113 sleeved on the rotating shaft 930 and the bottom blade structure 133 sleeved on the idle shaft 230 also rotate synchronously. With such an arrangement, a liquid flow driving force can be provided for the liquid in the microfluidic chip 10 by only driving the motor 920 in one step, and the ambient temperatures in the top temperature control unit 110 and the bottom temperature control unit 130 can be adjusted.
Referring to fig. 7, the amplification temperature control mechanism 300 includes a support frame 350, and a driving module 370, an upper temperature control module 310 and a lower temperature control module 330 disposed on the support frame 350, wherein the driving module 370 is configured to enable the upper temperature control module 310 and the lower temperature control module 330 to move towards each other to be clamped at two sides of the PCR amplification chamber of the microfluidic chip 10 or move away from the PCR amplification chamber in a back-to-back manner. The upper temperature control module 310 comprises a first heat radiator 311, a first semiconductor chilling plate 312 and a first heat conductor 313 which are sequentially connected in a stacked manner, and the lower temperature control module 330 comprises a second heat radiator 331, a second semiconductor chilling plate 332 and a second heat conductor 333 which are sequentially connected in a stacked manner. The first thermal conductor 313 is provided with a receiving groove (not shown), and the receiving groove is used for receiving a PCR amplification cavity protruding from the surface of the microfluidic chip. When the temperature of the PCR amplification cavity is controlled, the first heat conductor 313 and the second heat conductor 333 are enclosed at two sides of the PCR amplification cavity, and the PCR amplification cavity is located in the accommodating groove, so that the PCR amplification cavity is completely wrapped, rapid temperature adjustment is realized in a targeted manner, and the reaction time is saved. It can be understood that when the PCR amplification chamber protrudes from the other side surface of the microfluidic chip 10, the accommodating groove is disposed on the second thermal conductor 333.
The first and second heat sinks 311 and 331 may include heat sinks and fans, and the first and second heat conductors 313 and 333 may be made of metal.
For ease of viewing, the support 350 of the amplification temperature control mechanism 300 is omitted from FIG. 8. The drive module 370 includes a horizontal slider rail 320, a vertical slider rail 340, an upper link 375, and a lower link 376. The vertical slider rail 340 includes a vertical rail 341, an upper slider 342 and a lower slider 343 slidably coupled to the vertical rail 341. The vertical guide rail 341 is fixed on the supporting frame 350, the upper temperature control module 310 is connected with the upper slider 342, and the lower temperature control module 330 is connected with the lower slider 343. The horizontal slider rail 320 includes a cross rail 321, and a front slider 322 slidably coupled to the cross rail 321. The horizontal guide rail 321 is fixed on the supporting frame 350, and the length direction of the horizontal guide rail 321 is perpendicular to the length direction of the vertical guide rail 341. One end of the upper link 375 is hinged to the upper slider 342, the other end is hinged to the front slider 322, one end of the lower link 376 is hinged to the lower slider 343, and the other end is hinged to the front slider 322. When the front slider 322 slides along the horizontal rail 321, the upper link 375 is driven to drive the upper slider 342 to slide along the vertical rail 341, and the lower link 376 is driven to drive the lower slider 343 to slide along the vertical rail 341, so as to enable the upper temperature control module 310 and the lower temperature control module 330 to approach each other or move away from each other.
The required travel of the upper and lower temperature control modules 310, 330 can be accommodated by varying the length of the upper and lower links 375, 376.
In order to make the upper and lower temperature control modules 310 and 330 move smoothly, the vertical slider rails 340 are arranged in two symmetrical groups, and the upper and lower links 375 and 376 are also arranged in two symmetrical groups, and accordingly, a rear slider 323 is slidably disposed on the cross rail 321 to be hinged to the other group of the upper and lower links 375 and 376.
Referring to fig. 8, in the present embodiment, the amplification temperature control mechanism 300 further includes a cam mechanism and an elastic member 360, the cam mechanism includes a transmission shaft 371 and a cam 373 sleeved on the transmission shaft 371, and the cam 373 abuts against the upper slide block 342. One end of the elastic member 360 is connected to the upper temperature control module 310, and the other end is connected to the lower temperature control module 330. When the amplification temperature control mechanism 300 is not in operation, the elastic member 360 is in an undeformed state.
When the cam 373 rotates synchronously with the transmission shaft 371, the upper slider 342 slides along the vertical slider guide 340, and at the same time, the upper slider 342 drives the front slider 322 to slide along the transverse guide 321 through the upper link 375, and the lower link 376 connected with the front slider 322 also drives the lower slider 343 to slide along the vertical slider guide 340. In a series of linkage processes, the upper temperature control module 310 and the lower temperature control module 330 gradually move away from or close to each other; the elastic member 360 is also stretched as the upper and lower temperature control modules 310 and 330 are separated, and gradually restores its shape as the upper and lower temperature control modules 310 and 330 approach each other.
The elastic member 360 is disposed between the upper temperature control module 310 and the lower temperature control module 330, so as to ensure that the cam 373 is always attached to the upper slider 342. In other embodiments, a roller may be disposed between the cam 373 and the upper slider 342, and the roller may rotate relative to the upper slider 342 to reduce friction between the cam 371 and the upper slider 342.
Referring to fig. 9, the detecting mechanism 500 includes an optical fiber type fluorescence detector, the optical fiber type fluorescence detector includes a fixed disk 510 and a rotating disk 530, which are stacked, the fixed disk 510 is provided with an excitation light fiber 511 and an emission light fiber 512, and the rotating disk 530 is provided with an excitation light source module 531 and a fluorescence detecting module 532. One end of the excitation light fiber 511 is coupled to the excitation light source module 531, and the other end is connected to the first thermal conductor 313 in the amplification temperature control mechanism 300. One end of the emission optical fiber 512 is coupled to the fluorescence detection module 532, and the other end is connected to the first thermal conductor 313 in the amplification temperature control mechanism 300. The excitation light source module 531 is used for emitting excitation light, the excitation light is projected to the PCR amplification chamber through the excitation light fiber 511, the fluorescence is generated after the fluorescence in the PCR amplification chamber is excited, and the generated fluorescence is transmitted to the fluorescence detection module 532 through the emission light fiber 512 for processing to obtain a detection result.
In the present embodiment, the excitation-light source modules 531 and the fluorescence detection modules 532 are provided in three groups and are uniformly arranged in the circumferential direction of the rotating disk 530. By rotating the rotating disc 530, different groups of excitation light source modules 531 and fluorescence detection modules 532 can form different optical detection channels with the excitation optical fibers 511 and the emission optical fibers 512, so as to improve the detection efficiency.
According to the nucleic acid analysis device, in the nucleic acid extraction process, the extraction temperatures of different extraction areas of the microfluidic chip 10 are respectively controlled, and the temperature influence between adjacent extraction areas is reduced. In the present application, the top fan blade structure 113 and the bottom fan blade structure 133 are both rotated synchronously with the rotating shaft 930, and the air temperature above the extraction area of the microfluidic chip 10 can be adjusted by using one driving motor 920. Further, the top blade structure 113 drives air to form a dynamic airflow during rotation, and the airflow circulates only inside the top heat insulating cover 111, and similarly, the airflow formed during rotation of the bottom blade structure 133 also circulates only inside the bottom heat insulating cover 131, so that the temperatures of the air above the adjacent extraction regions do not affect each other while the temperatures in the top heat insulating cover 111 and the bottom heat insulating cover 131 are kept balanced. When the PCR amplification cavity carries out amplification reaction, the PCR amplification cavity is completely wrapped by the amplification temperature control mechanism, and the reaction temperature of the PCR amplification cavity can be quickly adjusted due to the large heating area and the short temperature transmission path of the PCR amplification cavity. The detection mechanism 500 is provided with a plurality of groups of optical detection channels, and all detection results can be obtained by rotating the rotating disc 530. Meanwhile, the corresponding groups of excitation light source modules 531 and fluorescence detection modules 532 can be arranged on the rotating disk 530 as required, so that the expansion is convenient. According to the nucleic acid analysis device, the whole nucleic acid detection process can be automatically controlled to be completed after the micro-fluidic chip 10 is placed in the bracket 23, and convenience and rapidness are achieved.
The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. A nucleic acid analysis device for analyzing a sample in a microfluidic chip, the microfluidic chip including an extraction region and an amplification chamber, the extraction region including a plurality of extraction temperature zones, comprising:
the centrifugal driving mechanism is used for driving the micro-fluidic chip to rotate and comprises a rotating shaft, and the rotating shaft is used for being connected with the micro-fluidic chip;
the extraction temperature control mechanism comprises a top temperature control unit and a bottom temperature control unit which are positioned at two sides of the microfluidic chip and are used for respectively controlling the air temperature of different extraction temperature areas on the microfluidic chip;
the amplification temperature control mechanism is positioned beside the extraction temperature control mechanism and comprises an upper temperature control module and a lower temperature control module, and the upper temperature control module and the lower temperature control module are used for sealing the amplification cavity and regulating the temperature of the amplification cavity when the amplification cavity carries out amplification reaction;
and the detection mechanism forms an optical detection channel with the amplification cavity through the amplification temperature control module.
2. The nucleic acid analysis device of claim 1, wherein the top temperature control unit comprises a top thermal shield and a top heating element, the top heating element is located in the top thermal shield, the bottom temperature control unit comprises a bottom thermal shield and a bottom heating element, the bottom heating element is located in the bottom thermal shield, and both the top thermal shield and the bottom thermal shield are not in contact with the microfluidic chip when the microfluidic chip is connected to the spindle.
3. The apparatus according to claim 2, wherein the top temperature control unit further comprises a top fan structure and a top deflector located in the top thermal shield, the top fan structure being sleeved on the shaft and located on one side of the top deflector; the bottom temperature control unit further comprises a bottom fan blade structure, a bottom flow guide disc and an idle shaft, wherein the bottom fan blade structure, the bottom flow guide disc and the idle shaft are located in the bottom heat insulation cover, the bottom fan blade structure is sleeved on the idle shaft and located on one side of the bottom flow guide disc, and the idle shaft is used for being connected with the rotating shaft to rotate synchronously along with the rotating shaft.
4. The nucleic acid analysis device of claim 3, wherein the top baffle and the bottom baffle are each provided with an air inlet and at least one air outlet therethrough.
5. The nucleic acid analysis device of claim 3, wherein the top deflector has a through air inlet in the central region and a through air outlet at the edge of the top deflector; the central area of the bottom flow guide disc is provided with a through central air inlet, and the bottom flow guide disc is provided with an inner ring air outlet and an outer ring air outlet along the radial direction.
6. The nucleic acid analysis device of claim 5, wherein the bottom baffle comprises a first air distribution plate and a second air distribution plate that are stacked, and the first air distribution plate and the second air distribution plate are relatively rotatable to communicate one of the inner ring air outlet and the outer ring air outlet with the central air inlet.
7. The apparatus according to claim 1, wherein a receiving chamber is provided on the upper temperature control module or the lower temperature control module, and the receiving chamber is configured to receive the amplification chamber.
8. The nucleic acid analysis device of claim 1, wherein the amplification temperature control mechanism further comprises a driving module, the driving module comprises an upper connecting rod, a lower connecting rod, and a vertically arranged horizontal slider rail and a vertical slider rail, the vertical slider rail comprises an upper slider connected to the upper temperature control module, and a lower slider connected to the lower temperature control module, one end of the upper connecting rod is hinged to the upper slider, the other end of the upper connecting rod is hinged to a slider on the horizontal slider rail, one end of the lower connecting rod is hinged to the lower slider, and the other end of the lower connecting rod is hinged to a slider on the horizontal slider rail.
9. The apparatus according to claim 8, wherein the driving module further comprises a transmission shaft and a cam sleeved on the transmission shaft, the cam abuts against the upper temperature control module, and the upper temperature control module and the lower temperature control module can move toward or away from each other during rotation of the cam.
10. The apparatus according to claim 9, wherein the amplification temperature control mechanism further comprises an elastic member, one end of the elastic member is connected to the upper temperature control module, the other end of the elastic member is connected to the lower temperature control module, the elastic member is in a deformed state when the upper temperature control module and the lower temperature control module are away from each other, and the elastic member is in an original state when the upper temperature control module and the lower temperature control module are in contact with each other.
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| CN202210036525.5A CN114177963A (en) | 2022-01-13 | 2022-01-13 | Nucleic acid analysis device |
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