CN219792989U - Nucleic acid amplification device - Google Patents
Nucleic acid amplification device Download PDFInfo
- Publication number
- CN219792989U CN219792989U CN202320882154.2U CN202320882154U CN219792989U CN 219792989 U CN219792989 U CN 219792989U CN 202320882154 U CN202320882154 U CN 202320882154U CN 219792989 U CN219792989 U CN 219792989U
- Authority
- CN
- China
- Prior art keywords
- temperature
- heating
- cooling
- heating mechanism
- nucleic acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000003199 nucleic acid amplification method Methods 0.000 title claims abstract description 79
- 230000003321 amplification Effects 0.000 title claims abstract description 66
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 62
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 62
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 62
- 238000010438 heat treatment Methods 0.000 claims abstract description 439
- 230000007246 mechanism Effects 0.000 claims abstract description 411
- 238000001816 cooling Methods 0.000 claims abstract description 202
- 238000006243 chemical reaction Methods 0.000 claims abstract description 128
- 239000004020 conductor Substances 0.000 claims abstract description 3
- 238000001514 detection method Methods 0.000 claims description 88
- 239000002826 coolant Substances 0.000 claims description 20
- 238000002791 soaking Methods 0.000 claims description 16
- 238000005507 spraying Methods 0.000 claims description 5
- 230000004308 accommodation Effects 0.000 claims 1
- 230000000630 rising effect Effects 0.000 abstract description 18
- 238000012546 transfer Methods 0.000 abstract description 10
- 238000000034 method Methods 0.000 description 71
- 238000009529 body temperature measurement Methods 0.000 description 37
- 230000008569 process Effects 0.000 description 34
- 230000008859 change Effects 0.000 description 20
- 238000000137 annealing Methods 0.000 description 18
- 238000004925 denaturation Methods 0.000 description 16
- 230000036425 denaturation Effects 0.000 description 16
- 238000005057 refrigeration Methods 0.000 description 11
- 238000004321 preservation Methods 0.000 description 10
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 239000000463 material Substances 0.000 description 5
- 229910052802 copper Inorganic materials 0.000 description 4
- 239000010949 copper Substances 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000007921 spray Substances 0.000 description 4
- 238000004861 thermometry Methods 0.000 description 4
- 229910052782 aluminium Inorganic materials 0.000 description 3
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 3
- 239000000919 ceramic Substances 0.000 description 3
- 238000005265 energy consumption Methods 0.000 description 3
- 229910052751 metal Inorganic materials 0.000 description 3
- 239000002184 metal Substances 0.000 description 3
- 238000003752 polymerase chain reaction Methods 0.000 description 3
- 238000004904 shortening Methods 0.000 description 3
- 238000010792 warming Methods 0.000 description 3
- 230000001419 dependent effect Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000020169 heat generation Effects 0.000 description 2
- 238000010030 laminating Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 239000004065 semiconductor Substances 0.000 description 2
- 238000003466 welding Methods 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 1
- 241001652065 Trigonopeltastes delta Species 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000000429 assembly Methods 0.000 description 1
- 230000000712 assembly Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 239000000110 cooling liquid Substances 0.000 description 1
- 238000012937 correction Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000000994 depressogenic effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000017525 heat dissipation Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001746 injection moulding Methods 0.000 description 1
- 239000011810 insulating material Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 239000007769 metal material Substances 0.000 description 1
- 239000002985 plastic film Substances 0.000 description 1
- 229920006255 plastic film Polymers 0.000 description 1
- 229920001296 polysiloxane Polymers 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
Landscapes
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The utility model relates to a nucleic acid amplification device, which comprises a cooling mechanism and a heating mechanism, wherein the heating mechanism is provided with a containing cavity, the containing cavity is of a flat structure, and the surface, close to the cooling mechanism, of the heating mechanism is formed by a heat conducting material. The holding cavity is of a flat structure, the reaction sample arranged in the holding cavity can be very thin, and the temperature on the surface of the reaction sample can be quickly transferred to the center, so that the temperature rising and reducing speed of the reaction sample is greatly improved. The holding cavity is directly formed on the heating mechanism, so that a heat transfer interface can be reduced, and the temperature rising and reducing speed of the reaction sample is further improved.
Description
Technical Field
The utility model relates to the technical field of medical treatment, in particular to a nucleic acid amplification device.
Background
PCR (polymerase chain reaction) is a molecular biological experimental method for in vitro enzymatic synthesis of specific DNA fragments, and PCR amplification, namely nucleic acid amplification, mainly comprises three repeated thermal cycles of high-temperature denaturation, low-temperature annealing and temperature-adaptive extension.
In the prior art, the new crown detection needs to keep warm for 10s-20s in three stages of high temperature denaturation, low temperature annealing and temperature adaptation extension, one-time heating or cooling needs to be about 30s, and the cycle needs to be 40-50 times, and the whole detection period needs about 1 hour. However, the existing nucleic acid amplification method needs to wait for a long time, and cannot meet the demands of users.
Disclosure of Invention
A nucleic acid amplification apparatus includes a cooling mechanism and a heating mechanism, the heating mechanism being formed with a housing chamber, the housing chamber being of a flat configuration.
Optionally, the cooling medium of the cooling mechanism is in direct contact with the heating mechanism for cooling.
Optionally, at least part of the upper surface of the heating means, or at least part of the upper surface of the recess of the heating means, acts as a bottom wall of the receiving cavity.
Optionally, the upper surface of the heating mechanism includes a soaking layer in direct contact with the reaction sample in the receiving cavity.
Optionally, the heating mechanism comprises a mechanism body and a bottom wall of the accommodating cavity;
the bottom wall is closely contacted with at least part of the upper surface of the mechanism body, or the bottom wall is closely contacted with the upper surface of the recess of the mechanism body, or
A flexible heat conducting element is arranged between the bottom wall and the mechanism body.
Optionally, at least two independent subchambers are arranged in the accommodating chamber; or the number of the accommodating cavities is one or at least two.
Optionally, the heating mechanism includes a heating body and a temperature calibration part for reflecting the temperature of the heating body.
Optionally, the nucleic acid amplification apparatus may further include a resistance detection member for detecting a resistance of the heating body.
Optionally, the heating mechanism further comprises an upper conduction assembly and a lower conduction assembly, and the heating body is sandwiched between the upper conduction assembly and the lower conduction assembly.
Optionally, the heating mechanism further includes a rapid conduction portion for conducting heat of the heating body to the temperature calibration portion.
Optionally, one side of the rapid conduction part is connected to one side of the upper conduction assembly close to the heating body or connected to one side of the lower conduction assembly close to the heating body, and the other side of the rapid conduction part is connected to the temperature calibration part.
Optionally, the temperature calibration part is located at the bottom of the heating mechanism.
Optionally, a receiving groove is formed in one side, far away from the heating body, of the lower conduction assembly, and the temperature calibration part is located in the receiving groove and connected with the bottom of the receiving groove.
Optionally, the quick conduction part includes one or more first guide posts, one end of the one or more first guide posts is attached to one side of the upper conduction assembly close to the heating body or attached to one side of the lower conduction assembly close to the heating body, and the other end of the one or more first guide posts is connected with the temperature calibration part.
Optionally, the quick conduction portion includes paster and one or more second guide pillar, the paster with go up the conduction subassembly is close to one side laminating of heating member or with lower conduction subassembly is close to one side laminating of heating member, one end of one or more second guide pillar connect in the paster, the other end wear to locate lower conduction subassembly and with temperature calibration portion connects.
Optionally, the lower conductive component is provided with a third through hole, and the second guide pillar is disposed in the third through hole.
Optionally, the nucleic acid amplification apparatus further comprises a temperature detection unit for detecting a temperature of the temperature calibration part.
Optionally, the cooling mechanism is provided with an avoiding portion for avoiding the temperature calibration portion and the temperature detection unit.
Optionally, the cooling mechanism includes the cooling main part and with the cold head that the cooling main part is connected, heating mechanism can paste tightly the cold head, the cooling main part can be for the cold head cooling.
Optionally, the cooling body cools the coldhead by spraying a cooling medium; or alternatively, the first and second heat exchangers may be,
the cooling medium for cooling the cold head flows through the cooling main body.
Optionally, the nucleic acid amplification apparatus further comprises a temperature control unit.
From the above, the accommodating cavity of the nucleic acid amplification device provided by the utility model is of a flat structure, the reaction sample arranged in the accommodating cavity can be very thin, and the temperature on the surface of the reaction sample can be quickly transferred to the center, so that the heating and cooling speed of the reaction sample is greatly improved. The holding cavity is directly formed on the heating mechanism, so that a heat transfer interface can be reduced, and the temperature rising and reducing speed of the reaction sample is further improved.
Drawings
FIG. 1 is a flow chart of a nucleic acid amplification method according to an embodiment of the present utility model;
FIG. 2 is a graph showing the comparison of the trend of temperature change of the water layers of continuous refrigeration and indirect refrigeration provided by the embodiment of the utility model;
FIG. 3 is a flowchart showing a method for amplifying nucleic acid according to an embodiment of the present utility model;
FIG. 4 is a graph showing the temperature profile of a reaction sample provided by an embodiment of the present utility model;
FIG. 5 is a schematic view of a first heating mechanism according to an embodiment of the present utility model;
FIG. 6 is a schematic diagram of a nucleic acid amplification apparatus according to an embodiment of the present utility model;
FIG. 7a is a schematic diagram showing the structure of another nucleic acid amplification apparatus according to an embodiment of the present utility model;
FIG. 7b is a schematic diagram showing the structure of another nucleic acid amplification apparatus according to an embodiment of the present utility model;
FIG. 8 is a schematic diagram showing the structure of another nucleic acid amplification apparatus according to an embodiment of the present utility model;
FIG. 9 is a cross-sectional view of one of the nucleic acid amplification apparatuses according to the embodiment of the present utility model;
FIG. 10 is a schematic view of a second heating mechanism according to an embodiment of the present utility model;
FIG. 11 is a schematic view of a third heating mechanism according to an embodiment of the present utility model;
FIG. 12 is a schematic view of a fourth heating mechanism according to an embodiment of the present utility model;
FIG. 13 is a schematic view of a fifth heating mechanism according to an embodiment of the present utility model;
fig. 14 is a schematic structural view of a sixth heating mechanism according to an embodiment of the present utility model;
fig. 15 is a schematic structural view of a cooling mechanism according to an embodiment of the present utility model.
In the figure:
1. a cooling mechanism; 11. a cooling flow passage; 12. a heat conductive plate; 13. a cooling body; 14. a cold head; 15. an avoidance unit;
2. a heating mechanism; 21. a receiving chamber; 22. a bottom wall; 23. a heating body; 24. an upper conductive assembly; 241. a soaking layer; 25. a temperature calibration unit; 251. an external electrical connection contact; 252. an electrical connection lead; 26. a fast conduction part; 261. a patch; 262. a second guide post; 263. a first guide post; 27. a lower conductive assembly; 271. an insulating thermal resistance layer; 273. a first through hole; 275. a receiving groove; 29. a flexible heat conducting member; 20. a second contact;
3. A first contact temperature detection unit; 4. a second contact temperature detection unit; 5. a resistance detecting member; 6. a non-contact temperature detection unit.
Detailed Description
The technical scheme of the utility model is further described below by the specific embodiments with reference to the accompanying drawings. It is to be understood that the specific embodiments described herein are merely illustrative of the utility model and are not limiting thereof. It should be further noted that, for convenience of description, only some, but not all of the drawings related to the present utility model are shown.
In the present utility model, directional terms such as "upper", "lower", "left", "right", "inner" and "outer" are used for convenience of understanding, and thus do not limit the scope of the present utility model unless otherwise specified.
In the present utility model, unless expressly stated or limited otherwise, a first feature "above" or "below" a second feature may include both the first and second features being in direct contact, as well as the first and second features not being in direct contact but being in contact with each other through additional features therebetween. Moreover, a first feature being "above," "over" and "on" a second feature includes the first feature being directly above and obliquely above the second feature, or simply indicating that the first feature is higher in level than the second feature. The first feature being "under", "below" and "beneath" the second feature includes the first feature being directly under and obliquely below the second feature, or simply means that the first feature is less level than the second feature.
In the description of the present utility model, unless explicitly stated and limited otherwise, the terms "connected," "connected," and "fixed" are to be construed broadly, and may be, for example, fixedly connected, detachably connected, or integrally formed; can be mechanically or electrically connected; can be directly connected or indirectly connected through an intermediate medium, and can be communicated with the inside of two elements or the interaction relationship of the two elements. The specific meaning of the above terms in the present utility model will be understood in specific cases by those of ordinary skill in the art.
Example 1
The present embodiment provides a nucleic acid amplification method for use in a new crown detection, but is not limited to this, and may be used in other detection to improve the detection efficiency and reduce the time required for the detection.
The nucleic acid amplification method provided in this embodiment includes keeping the reaction sample continuously refrigerated, and controlling the temperature of the reaction sample by controlling the heating mechanism 2.
The continuous cooling of the reaction sample may be that the cooling mechanism 1 keeps continuous cooling so that the cooling mechanism 1 continuously cools the reaction sample, for example, the cooling mechanism 1 continuously cools the reaction sample indirectly, specifically, the reaction sample is placed in the heating mechanism 2, the cooling mechanism 1 is located at a side of the heating mechanism 2 away from the reaction sample, or the reaction sample is located at an upper side of the heating mechanism 2, the cooling mechanism 1 is located at a side of the heating mechanism 2 away from the reaction sample, and the cooling mechanism 1 further cools the reaction sample in the heating mechanism 2 by cooling the heating mechanism 2. For another example, the heating mechanism 2 is not arranged between the cooling mechanism 1 and the reaction sample, but directly refrigerates the reaction sample. For example, the reaction sample is heated by radiation, and the cooling mechanism 1 refrigerates at least one of the surfaces of the reaction sample.
This embodiment maintains the portion of the heating mechanism 2 close to the cooling mechanism 1 at a lower temperature at all times by keeping the reaction sample continuously refrigerated. When the heat of the reaction sample is transferred through the heating mechanism 2 to heat or cool the reaction sample, or the cooling mechanism 1 is located at one side of the heating mechanism 2 away from the reaction sample, the part of the heating mechanism 2, which is close to the cooling mechanism 1, is always kept at a lower temperature, so that when the reaction sample needs to be cooled, the cooling mechanism 1 only needs to cool the other part of the heating mechanism 2 and the reaction sample. That is, since the volume of the heating mechanism 2 to be cooled is reduced, the volume of the whole portion to be cooled is reduced, thereby shortening the cooling time.
In the cooling, heating, low-temperature annealing stage, high-temperature denaturation stage or temperature-adaptive extension stage, the heating mechanism 2 is controlled, so that the other part of the heating mechanism 2 close to the reaction sample reaches the required temperature by controlling the heating mechanism 2, the heat preservation, the rapid cooling and the rapid heating of the reaction sample are realized, the process of lifting and cooling the reaction sample is controlled within 2.5s, and the detection time is greatly shortened.
It is understood that when the heating mechanism 2 includes the heating body 23, the portion of the heating mechanism 2 close to the cooling mechanism 1 may be a portion of the heating mechanism 2 closer to the cooling mechanism 1 than the heating body 23. Another portion of the heating mechanism 2 may include a heating body 23 of the heating mechanism 2, and a portion farther from the cooling mechanism 1 than the heating body 23.
When the reaction sample is kept continuously refrigerated, the cooling mechanism 1 continuously refrigerates, and the cooling mechanism 1 is cooled in advance and is cooled for the heating mechanism 2 before the reaction sample needs to be cooled, so that the cooling mechanism can be in seamless connection with the cooling requirement of the reaction sample, and therefore, the cooling speed can be improved especially.
In a specific embodiment, as shown in fig. 2, a comparison curve of the trend of the temperature change of the water layer in the heating mechanism 2 of two different refrigeration modes, namely, the cooling mechanism 1 continuously refrigerates the reaction sample and the cooling mechanism intermittently refrigerates the reaction sample, is given. The nucleic acid high temperature denaturation temperature was assumed to be 95℃and the low temperature annealing temperature was assumed to be 55 ℃. For keeping the reaction sample continuously cooled, the cooling mechanism 1 is kept continuously in a low temperature state, for example, the temperature of the cooling mechanism 1 is kept at 5 ℃ all the time. In the case of intermittent cooling of the cooling mechanism 1, the cooling mechanism 1 is raised to the denaturation temperature or the cooling mechanism 1 does not actively cool any more, and then the cooling mechanism cools (e.g., down to 5 ℃) the heating mechanism 2 during the cooling stage of the reaction sample, so that the temperature of the sample is reduced to the annealing temperature. By finite element simulation analysis, comparing the cooling time in the above two cases, as shown in fig. 2, the cooling mechanism 1 keeps continuously cooling to 55 ℃ in advance of the intermittent cooling scheme by about 1.5 seconds. From this, the continuous refrigeration scheme is more advantageous to reduce the cooling time than the intermittent refrigeration scheme, thereby further reducing the nucleic acid amplification time.
Optionally, the nucleic acid amplification method comprises the steps of:
s0, keeping continuous refrigeration on a reaction sample; if the cooling mechanism 1 keeps continuously refrigerating, the cooling mechanism 1 continuously refrigerates the reaction sample;
step S1, a heating process is carried out, and a heating mechanism 2 is controlled to heat a reaction sample to a denaturation temperature or an extension temperature;
and S2, controlling the heating mechanism 2 to cool the reaction sample to the minimum value of the annealing temperature in the cooling process.
In the heating process and the cooling process, the cooling mechanism 1 only needs to keep continuously refrigerating, and only through the change of the heating mechanism 2, such as through controlling the power change of the heating mechanism 2, the part of the heating mechanism 2 close to the cooling mechanism 1 is always kept at a lower temperature in the heating process and the cooling process, so that when the reaction sample needs to be cooled, the cooling mechanism 1 only needs to cool the other part of the heating mechanism 2 and the reaction sample. That is, since the volume of the heating mechanism 2 to be cooled is reduced, the volume of the whole part to be cooled is reduced, thereby shortening the cooling time, and simultaneously, the cooling mechanism 1 does not need to perform pre-cooling and moving actions again, thereby achieving the purpose of rapid temperature rise and rapid temperature reduction of the reaction sample, and the control method is simple and has high control precision. Another reason is that since the portion of the heating mechanism 2 close to the cooling mechanism 1 is always maintained at a low temperature, when the reaction sample needs to be cooled down, the cooling mechanism 1 only needs to cool down the other portion of the heating mechanism 2 and the reaction sample. That is, since the volume of the heating mechanism 2 to be cooled is reduced, the volume of the whole portion to be cooled is reduced, thereby shortening the cooling time.
Specifically, the heat of the heating mechanism 2 is controlled to counter-charge the heat of the cooling mechanism 1, so that the reaction sample is warmed, cooled or kept at a temperature. The heat quantity of the cooling mechanism 1 may be opposed by controlling the heat quantity of the heating mechanism 2, and the heat quantity of the cooling mechanism 1 transferred to the heating mechanism 2 may be opposed by controlling the heat quantity of the heating mechanism 2. In the heating stage, the heat of the heating mechanism 2 is increased, for example, the heat generation amount of the heating mechanism 2 in unit time is increased, the heat transferred to the heating mechanism 2 by the cooling mechanism 1 is opposed to the part of the heat of the heating mechanism 2, so that the rapid heating of the reaction sample is realized, and therefore, the rapid heating of the reaction sample can be controlled by the heating mechanism 2, and the cooling mechanism 1 continuously refrigerates the reaction sample and does not influence the rapid heating of the reaction sample. In the cooling stage, the heat of the heating mechanism 2 is reduced, for example, the heating mechanism 2 reduces the heat generation amount per unit time, and the continuous refrigeration of the reaction sample and the control of the heating mechanism 2 by the cooling mechanism 1 realize the rapid cooling of the heating mechanism 2 and the reaction sample. While maintaining the temperature, the heat of the heating mechanism 2 is controlled according to the detected temperature of the reaction sample, and the heat transferred to the heating mechanism 2 by the cooling mechanism 1 is further opposed to the reaction sample so as to maintain the reaction sample at a desired temperature.
Alternatively, the temperature of the reaction sample is controlled by controlling the power of the heating mechanism 2. The temperature of the heating mechanism 2 can be controlled by controlling the power of the heating mechanism 2, so that the temperature of the reaction sample is controlled, the control method is simple, and the control precision is high. Because the power is the product of the current and the voltage, the temperature of the reaction sample can be controlled by controlling the current of the heating mechanism 2.
Alternatively, the heating means 2 is operated at a higher power during the warming up than during the cooling down of the heating means 2.
In the heating process, the heating mechanism 2 operates at high power to offset the low temperature of the cooling mechanism 1, so that the nucleic acid amplification method with continuous refrigeration can achieve the purpose of rapid heating by the high-power operation of the heating mechanism 2 relative to intermittent refrigeration.
During the temperature decrease, the heating mechanism 2 is operated at a low power or the heating mechanism 2 stops heating. Wherein the heating mechanism 2 is operated at a low power to avoid the temperature of the reaction sample from falling below the desired temperature.
Optionally, as shown in fig. 1, the nucleic acid amplification method includes the steps of:
s0, keeping continuous refrigeration on the sample;
step S1 is executed, so that the temperature of the reaction sample is raised to the denaturation temperature;
S3, in a high-temperature denaturation stage, controlling the heating mechanism 2 to enable the reaction sample to be kept at a denaturation temperature for a first preset time;
executing the step S2;
s4, in the low-temperature annealing stage, the heating mechanism 2 is controlled to keep the reaction sample at the annealing temperature for a second preset time;
step S1 is executed, so that the reaction sample is heated to an extension temperature;
s5, in the temperature-adaptive extension stage, the heating mechanism 2 is controlled to keep the reaction sample at the extension temperature for a third preset time;
the steps are circulated for a plurality of times until a preset number of cycles or a preset amplification level is reached.
In the heat preservation, heating up process and cooling down process, the cooling mechanism 1 is required to keep continuous refrigeration on the reaction sample, and the temperature of the heating mechanism 2 is changed only through the change of the heating mechanism 2, such as the power change or the current change of the heating mechanism 2, so that the purposes of heat preservation, rapid heating up and rapid cooling down of the reaction sample are achieved, and the control method is simple. In the heat preservation stage, the power or current of the heating mechanism 2 is basically kept unchanged; in the temperature lowering stage, the power or current of the heating mechanism 2 is reduced to lower the temperature of the heating mechanism 2; in the temperature rising stage, the power or current of the heating mechanism 2 is increased to raise the temperature of the heating mechanism 2. Specifically, the heating means 2 is heated by resistance heating, and the heating power and thus the temperature of the heating means 2 are controlled by controlling the current and/or voltage value of the resistance. The power control of the heating means 2 can be achieved typically by means of a controllable current source, a controllable voltage source, or by means of a combination of constant voltage and current switches.
Alternatively, the temperature of the cooling mechanism 1 is opposed by controlling the temperature of the heating mechanism 2 so that the reaction sample is maintained at the denaturation temperature, the annealing temperature or the extension temperature. For example, the power of the heating mechanism 2 is controlled, so that the purpose of controlling the temperature of the heating mechanism 2 is achieved.
Alternatively, the heating mechanism 2 is pulse heating. The heating speed of pulse heating is faster, and the temperature control is easier. Of course, in other alternative embodiments, the heating mechanism 2 may also employ resistive wire heating, electromagnetic heating, or microwave heating. Or the heating mechanism 2 comprises a heat conducting film such as an aluminum plastic film and a resistance wire arranged in the heat conducting film, and after the resistance wire generates heat, the heat conducting film can rapidly transfer heat to a reaction sample, so that the heat conducting efficiency is improved.
Optionally, the sample is kept continuously refrigerated: the cooling mechanism 1 continues to cool the heating mechanism 2, and the temperature of the cooling mechanism 1 is not higher than the minimum value of the annealing temperature. The cooling mechanism 1 continuously refrigerates the heating mechanism 2 to continuously cool the heating mechanism 2 and further cool the reaction sample placed in the heating mechanism 2.
The cooling mechanism 1 is always used for refrigerating the heating mechanism 2, on one hand, when a reaction sample needs to be cooled, the heating power of the heating mechanism 2 is reduced, so that the temperature distribution reaches new balance, and the purpose of cooling is achieved, and the cooling structure 1 only needs to cool the reaction sample and part of the heating mechanism 2, so that the cooling time is shortened. Meanwhile, when cooling is needed, the cold area mechanism can immediately cool the heating mechanism 2, so that the cooling speed of the heating mechanism 2 is increased; in addition, when the cooling mechanism 1 can adopt a contact type mode to cool the heating mechanism 2, the cooling mechanism 1 is a contact type cooling mechanism which is used for cooling by contacting with the heating mechanism 3, so that the cooling mechanism 1 does not need to reciprocate, a driving mechanism for driving the cooling mechanism 1 to move is omitted, the cost is reduced, and the control method is simplified; meanwhile, the cooling mechanism 1 does not need to repeatedly contact and separate from the heating mechanism 2, so that the impact on the heating mechanism 2 caused by the contact of the cooling mechanism 1 with the heating mechanism 2 is avoided, the stability of the heating mechanism 2 is improved, and the influence on a reaction sample on the heating mechanism 2 is avoided. The temperature of the cooling mechanism 1 is not higher than the annealing temperature, and since the annealing temperature is the lowest temperature of the reaction sample in the temperature raising and lowering cycle, the temperature of the cooling mechanism 1 is not higher than the annealing temperature, the cooling mechanism 1 is always at a lower temperature, so that the reaction sample can be cooled at any time in the temperature raising and lowering cycle, and the cooling mechanism 1 is in a lower temperature state, thereby accelerating the cooling of the heating mechanism 2 and the cooling of the reaction sample placed on the heating mechanism 2. In an alternative embodiment, the temperature of the cooling means 1 is not higher than the annealing temperature, in particular the temperature of the cooling means 1 is substantially the same temperature, e.g. about 5 ℃, so that the temperature of the cooling means 1 is continuously maintained at a temperature that can rapidly cool down the heating means 2.
Considering that the temperature of the cooling mechanism 1 is continuously maintained at a temperature that can rapidly cool down the heating mechanism 2, which increases the energy consumption of the heating mechanism 2 during the temperature rising process and the temperature maintaining process, in other alternative embodiments, the cooling mechanism 1 continuously cooling the heating mechanism 2 may include: the cooling mechanism 1 is continuously contacted with the heating mechanism 2, and the cooling mechanism 1 is at a first temperature in the cooling process; in the heating process, the cooling mechanism 1 is at a second temperature, in the temperature-adaptive extension stage, the cooling mechanism 1 is at a third temperature, in the high-temperature denaturation stage, the cooling mechanism 1 is at a fourth temperature, and the second temperature, the third temperature and the fourth temperature are not lower than the first temperature.
The second temperature, the third temperature, and the fourth temperature may be equal or different. For example, the first temperature may be 5 °, the second, third and fourth temperatures may be other temperatures above 5 ℃, such as 10 °, 20 °, etc. The second temperature, the third temperature and the fourth temperature are not lower than the first temperature, and especially when the second temperature, the third temperature and the fourth temperature are higher than the first temperature, the heat dissipation of the heating mechanism 2 can be reduced, and the energy consumption of the heating mechanism 2 can be further reduced.
Further, in order to further accelerate the cooling efficiency, the cooling mechanism 1 continuously cools the heating mechanism 2 further includes: before the reaction sample is cooled, the temperature of the cooling mechanism 1 is reduced to the first temperature to be prepared in advance for cooling the heating mechanism 2 and the reaction sample. It will be appreciated that the temperature of the reaction sample can be controlled by controlling the heating means 2 in order to maintain the temperature of the reaction sample after the cooling means 1 has cooled down before the reaction sample has cooled down. Such as by increasing the power of the heating mechanism 2, etc., to maintain the desired temperature of the reaction sample.
Alternatively, the cooling mechanism 1 may adopt a contact type manner to cool the heating mechanism 2 and the reaction sample, specifically, the contact type is: the heat exchange is realized by contact between the cooling mechanism 1 and the heating mechanism 2, for example, the cooling mechanism 1 comprises a semiconductor refrigerator (refer to fig. 7 a) contacted with the heating mechanism 2 or comprises a heat conducting plate 12 (refer to fig. 7 b) contacted with the heating mechanism 2, the heat conducting plate 12 can be cooled by water cooling, air cooling and the like, and the semiconductor refrigerator or the heat conducting plate 12 is contacted with the heating mechanism 2, so that the temperature of the heating mechanism 2 is reduced. The heat conductive plate 12 may be made of metal or the like. The cooling mechanism 1 is continuously contacted with the heating mechanism 2 to continuously cool the heating mechanism 2, and further cool the reaction sample placed in the heating mechanism 2 or on the upper side of the heating mechanism 2.
As shown in fig. 6, of course, the cooling mechanism 1 may also adopt a non-contact manner to cool the heating mechanism 2 and the reaction sample, specifically, the non-contact manner is: the cooling mechanism 1 is not in contact with the heating mechanism 2, for example, the heating mechanism 2 may be cooled by a cooling medium. For example, the cooling means 1 may continuously spray water or a cooling medium such as gas to the heating means 2, or may continuously cool the heating means 2, thereby cooling the reaction sample placed in the heating means 2 or above the heating means 2.
In an alternative embodiment, the nucleic acid amplification method comprises the steps of:
controlling the heating mechanism 2, such as by increasing the power of the heating mechanism 2, to rapidly raise the temperature of the reaction sample to an optimal high temperature denaturation temperature, such as 95 ℃;
controlling the heating mechanism 2, such as by performing pulse width modulation-based switching control on the output power of the heating mechanism 2, so as to control the output average power, so that the reaction sample keeps stable temperature in a certain time in the high-temperature denaturation stage; or, the temperature of the reaction sample is kept stable in a certain time in the high-temperature denaturation stage by controlling the current or voltage of the heating mechanism 2;
controlling the heating mechanism 2, such as turning off the heating mechanism 2, to rapidly reduce the temperature of the reaction sample to an annealing temperature, such as 45 ℃;
Controlling the heating mechanism 2 to keep the temperature of the reaction sample stable in a certain time in the annealing stage;
controlling the heating mechanism 2, such as by increasing the power of the heating mechanism 2, to rapidly raise the temperature of the reaction sample to an extended temperature, such as 75 ℃;
the heating mechanism 2 is controlled to keep the temperature of the reaction sample stable in a certain time in the temperature-delay stage, and one cycle of nucleic acid amplification is completed.
The process is then cycled until detection is complete. It will be appreciated that the user sets the time for the reaction sample to be subjected to the high temperature denaturation stage, the low temperature annealing stage and the temperature elongation stage according to the reaction conditions.
As shown in fig. 3 and 4, in order to improve the accuracy of temperature control, optionally, the nucleic acid amplification method further includes: the temperature measurement is realized by a resistance temperature measurement method and a double temperature measurement method for calibrating the temperature of the heating mechanism 2.
In this embodiment, the heating mechanism 2 adopts a resistive heating mode to perform heating, that is, the heating mechanism 2 may include a heating body 23, and the heating body 23 may be a resistor. Since there is a specific relationship between the resistance and its temperature, the current corresponding temperature value can be obtained by measuring the resistance value of the heating mechanism 2. For the same type of resistor, such as a copper wire resistor, the resistance value at the nominal temperature between the resistors (the resistance value at the nominal temperature is simply referred to as the nominal resistance value), the nominal resistance means that the declared (or marked) resistance value is real at this temperature, wherein this temperature is the nominal temperature, the nominal temperature can be arbitrarily selected according to the requirement) and the temperature coefficient of resistance are slightly different, which can cause the difference between the temperature measured by the resistance temperature measurement method and the actual temperature, and the temperature of the heating mechanism 2 can be calibrated to correct the difference, so that the resistance temperature measurement method can determine the accurate temperature value.
Optionally, the temperature measurement by a resistance temperature measurement method and a dual temperature measurement method for calibrating the temperature of the heating mechanism 2 comprises the following steps:
the temperature calibration value of the heating mechanism 2 is measured by the temperature detection unit, and the temperature value obtained by the resistance thermometry value is calibrated by the temperature calibration value. That is, the temperature detecting unit detects the temperature calibration value of the heating mechanism 2, and the temperature measurement by the resistance temperature measuring method is calibrated by combining the temperature calibration value with the voltage and the current of the heating mechanism 2 detected by the resistance temperature measuring method.
Preferably, the temperature detecting unit detects the temperature at a temperature calibrating portion 25 (described in detail later) of the heating mechanism 2, thereby obtaining a temperature calibration value. It will be appreciated that the temperature calibration value is an artificially selected temperature value that is responsive to the current actual temperature of the heating means 2.
Specifically, the temperature coefficient of resistance and the nominal resistance value of the heating mechanism 2 are obtained from the temperature calibration value to calibrate the temperature value. The temperature calibration value is combined with the resistance value of the heating mechanism 2 under the temperature calibration value, so that the resistance temperature coefficient and the nominal resistance value corresponding to the specific resistance can be obtained, and the actual temperature of the heating mechanism 2 can be obtained by measuring the temperature through the resistance temperature coefficient and the nominal resistance value by using a resistance temperature measurement method.
Optionally, the timing of the temperature calibration value acquisition is: the temperature calibration is measured prior to the nucleic acid amplification process, during a first temperature increase of the nucleic acid amplification process, and/or during a first amplification cycle of the nucleic acid amplification process.
It is understood that the temperature detection unit detects the room temperature of the heating mechanism 2 as a temperature calibration value before the progress of nucleic acid amplification. It is also possible to make the heating means 2 have a certain temperature change before the nucleic acid amplification process to obtain a temperature calibration value by the temperature detecting unit, but this increases the detection time, and therefore it is preferable to measure the temperature calibration value in combination with the room temperature of the heating means 2 detected by the temperature detecting unit and during the first temperature increase of the nucleic acid amplification process and/or during the first amplification cycle of the nucleic acid amplification process.
It will be appreciated that the first warming up of the nucleic acid amplification process is the warming up to the pretreatment stage. The temperature rising process, the high-temperature denaturation stage, the cooling process, the low-temperature annealing stage, the temperature rising process and the temperature adapting extension stage are taken as a primary amplification cycle, and the temperature calibration value is measured during the primary amplification cycle. The time for acquiring the temperature calibration value provided by the embodiment can ensure the accuracy of subsequent temperature control.
Since the temperature change of the temperature detecting unit has a certain delay relative to the temperature change of the heating mechanism 2, after the temperature of the temperature detecting unit is consistent with the temperature of the heating mechanism 2, the temperature calibration value is read to ensure that the temperature calibration value reflects the current actual temperature of the heating mechanism 2. If the temperature calibration value is obtained in the temperature rising and reducing process, the speed of rising and reducing is slowed down so that the temperature of the temperature detection unit is consistent with the temperature of the heating mechanism 2, or the temperature change is suspended in the temperature rising and reducing process, and the temperature rising and reducing process is executed again after the temperature calibration value is obtained; if the temperature calibration value is obtained in the heat preservation stage, the temperature detected by the temperature detection unit is read after the heat preservation is carried out for a certain period of time, for example, the temperature detected by the temperature detection unit is read after the heat preservation is carried out for 2s, 3s or 8s, etc.
As shown in fig. 4, the heating mechanism 2 is optionally controlled in accordance with the temperature value detected by the resistance thermometry before the temperature value is calibrated. This can prevent the temperature of the heating mechanism 2 from being excessively deviated due to rapid temperature change of the heating mechanism 2. After the temperature value is calibrated, the temperature value detected by the resistance temperature measurement method is continued to control the heating mechanism 2. Of course, in other alternative embodiments, it is also possible to control the heating mechanism 2 in dependence of the temperature detected by the temperature detection unit before calibrating the temperature value.
Specifically, the temperature measurement is realized by a resistance temperature measurement method and a double temperature measurement method for calibrating the temperature of the heating mechanism 2, and the method comprises the following steps:
obtaining at least two different temperature calibration values; detecting a first voltage and a first current of the heating mechanism 2 at a temperature calibration value, and obtaining a first resistance of the heating mechanism 2 according to the first voltage and the first current;
obtaining the resistivity and nominal resistance of the heating mechanism 2 according to at least two first resistors;
the current and voltage of the heating mechanism 2 are continuously detected, and the temperature curve of the heating mechanism 2 is obtained according to the resistivity and the nominal resistance value.
It will be appreciated that the at least two different temperature calibrations obtained are read when the temperature detection unit substantially coincides with the temperature of the heating means 2, which temperature is considered to be the actual temperature of the heating means 2. At this time, the corresponding current and voltage of the heating mechanism 2 are read, and the resistivity and the nominal resistance value are reversely deduced, that is, calibration of the resistivity and the nominal resistance value is achieved, so that the temperature of the heating mechanism 2 can be considered to be the accurate temperature of the heating mechanism 2 when the temperature of the heating mechanism 2 is obtained from the current and voltage of the heating mechanism 2 later.
It will be appreciated that the greater the difference between the different temperature calibration values, the more accurate the resulting resistivity and nominal resistance values, and therefore, alternatively, the difference between adjacent two temperature calibration values is not less than 20 ℃.
According to the formula: according to the formula r=r 0 (1+αΔt) (wherein Δt=t-T 0 R is the corresponding resistance value of the heating body 23 at the temperature T, T 0 At nominal temperature, R 0 Is the nominal resistance value and α is the temperature coefficient of resistance of the material) to calibrate the temperature coefficient of resistance and the nominal resistance value of the heating mechanism 2. The formula can accurately obtain the relation between the temperature and the resistivity, the nominal resistance value, the voltage and the current. Specifically, the temperature detection unit detects a first temperature calibration value T 1 The resistance detection unit 5 detects that the heating body 23 is at T 1 First voltage U at temperature 1 And a first current I 1 From r=u/I, the heating body 23 can be obtained at T 1 Resistance R at temperature 1 The method comprises the steps of carrying out a first treatment on the surface of the Then the temperature detecting unit detects a second temperature calibration value T 2 The resistance detection unit 5 detects that the heating body 23 is at T 2 Second voltage U at temperature 2 And a second current I 2 From r=u/I, the heating body 23 can be obtained at T 2 Resistance R at temperature 2 Finally, according to two sets of binary once equations: r is R 1 ＝R 0 (1+αΔT 1 ) And R is 2 ＝R 0 (1+αΔT 2 ) (wherein DeltaT) 1＝ T 1 -T 0 ；ΔT 2＝ T 2 -T 0 ) To obtain alpha and R 0 Specific values of (3). By then continuously measuring the voltage and current of the heating means 2, according to the formula r=r 0 (1+αΔt) the temperature profile of the heating means 2 is obtained. Due to R corresponding to the specific heating body 23 in the formula 0 And alpha value, thus the can be quasiThe temperature value is obtained. It is understood that the resistance detection unit 5 may be any structure that can detect the current and voltage of the heating body 23.
In order to more clearly describe the nucleic acid amplification method in this embodiment, a process of calibrating a resistance thermometry by a temperature detection unit in an actual detection is shown in conjunction with FIG. 4. Before calibrating the temperature values, an initial RT temperature profile, i.e. a temperature preset profile, is preset, and then a small current, e.g. less than 1 milliamp, is applied to the heating body 23 of the heating means 2. In which a small current is applied for the purpose of reading the resistance of the heating body 23 without heating the heating body 23.
First calibration: the temperature detecting unit detects a first temperature calibration value T 1 The resistance detection unit detects that the heating body 23 is at T 1 First voltage U at temperature 1 And a first current I 1 From r=u/I, the heating body 23 can be obtained at T 1 Resistance R at temperature 1 。
Second calibration: then the temperature detecting unit detects a second temperature calibration value T 2 The resistance detection unit 5 detects that the heating body 23 is at T 2 Second voltage U at temperature 2 And a second current I 2 From r=u/I, the heating body 23 can be obtained at T 2 Resistance R at temperature 2 。
Finally, according to two sets of binary once equations: r is R 1 ＝R 0 (1+αΔT 1 ) And R is 2 ＝R 0 (1+αΔT 2 ) Obtaining R 0 And the specific value of alpha, an accurate R-T curve is obtained, and the temperature of the heating body 23 measured by a resistance temperature measurement method can be used as feedback for accurately controlling the temperature.
Obtaining R 0 And a by continuously measuring the voltage and current of the heating means 2 after a specific value of a, according to the formula r=r 0 (1+T.DELTA.T) the temperature profile of the heating mechanism 2 was obtained. Due to R corresponding to the specific heating body 23 in the formula 0 And an alpha value, so that the temperature value can be accurately obtained.
With continued reference to FIG. 4, upon acquisition of the first temperature calibration value, the temperature calibrationThe value can be considered as the nominal resistance value, and therefore, according to this temperature calibration value, the formula r=r 0 R in (1+αΔT) 0 And (3) performing primary correction to realize primary calibration on the temperature value measured by the resistance temperature measurement method (such as primary fluctuation of a temperature value curve measured by the resistance temperature measurement method in fig. 4 after primary calibration), then controlling the heating mechanism 2 to rise to a first stage of pretreatment by the temperature value obtained by the resistance temperature measurement method or the temperature value measured by the temperature detection unit, after the heating mechanism 2 is heated to the first stage of pretreatment for 8 seconds (not limited to 8 seconds, and can be any time longer than 2 seconds and shorter than the first-stage heat preservation time of pretreatment), obtaining a second temperature calibration value, performing secondary calibration on the temperature value detected by the resistance temperature measurement method according to the two temperature calibration values, controlling the heating mechanism 2 to be accurate temperature, and then continuously controlling the heating mechanism 2 according to the temperature value obtained by the resistance temperature measurement method. It will be appreciated that when the first temperature calibration value is room temperature, the temperature value measured by the resistance temperature measurement method may be calibrated for the first time, then a second temperature calibration value may be obtained at any temperature higher than room temperature, and the temperature value detected by the resistance temperature measurement method may be calibrated for the second time according to the two temperature calibration values, thereby completing the calibration of the temperature value.
The temperature calibration value can be detected in the whole process of nucleic acid amplification, so that the temperature can be calibrated for multiple times in the subsequent process, and the detection precision is further improved.
Optionally, at least two unequal temperature calibration values are measured, the temperature values being calibrated by the at least two unequal temperature calibration values. Such as measuring two, three, four or more temperature calibration values. The two temperature calibration values can be used for obtaining a resistivity and a nominal resistance value, the three or more temperature calibration values can be used for obtaining more than one resistivity and more than one nominal resistance value, and the more than one resistivity and more than one nominal resistance value can be used for obtaining a value which is closer to the actual resistivity and the nominal resistance value, so that the detection precision is further improved.
Since a certain time is required for transferring heat from the heating mechanism 2 to the temperature detecting unit, the temperature detecting unit can detect the temperature in 1-2s in the time of 1-2s in the rapid temperature rising and falling process, and the temperature change of the heating mechanism 2 can reach more than 30 ℃, so that the control of the heating mechanism 2 by the temperature detecting unit is relatively difficult in the rapid temperature rising and falling process. The resistance temperature measurement method can measure the real-time resistance change of the resistor while heating, and deduce the average temperature of the resistor through the temperature coefficient of the resistor and the nominal resistance. The temperature reflects the current temperature of the heating mechanism 2 in real time without delay, so that the temperature can be used for rapidly controlling the temperature of the sample. Since the temperature measurement method has the disadvantage that the temperature coefficient of resistance of a single resistor is slightly different from the nominal resistance value, temperature measurement errors may be caused. The embodiment does not depend on the temperature value measured by an uncalibrated resistance temperature measurement method completely, and also does not depend on the temperature detected by the temperature detection unit completely to control the heating mechanism 2, but the defects are overcome by combining the two, and the temperature of the heating mechanism 2 is calibrated, so that the temperature of the heating mechanism 2 can be controlled rapidly and accurately, and the aim of accurately controlling the temperature is fulfilled.
Example two
As shown in FIGS. 5 to 9, the present embodiment also provides a nucleic acid amplification apparatus. The nucleic acid amplification apparatus according to the second embodiment preferably performs the nucleic acid amplification method according to the first embodiment, and the nucleic acid amplification apparatus according to the second embodiment may be used without using the nucleic acid amplification method according to the first embodiment.
As shown in fig. 5, the nucleic acid amplification apparatus includes a cooling mechanism 1 and a heating mechanism 2, the heating mechanism 2 is formed with a housing chamber 21, the housing chamber 21 is of a flat structure, and a surface of the heating mechanism 2 adjacent to the cooling mechanism 1 is formed of a heat conductive material so that the heating mechanism 2 rapidly transfers heat to the cooling mechanism 1. It is to be understood that the flat structure may mean that the dimension in the thickness direction of the accommodating chamber 21 is much smaller than the dimension in the width or length direction, and as an example, the accommodating chamber 21 is a rectangular parallelepiped, and the ratio of the length and thickness of the rectangular parallelepiped may be greater than 5:1, such as 90:1, for example, the dimension in the thickness direction of the accommodating chamber 21 may be 0.3 to 1.0mm, and the width and length of the accommodating chamber 21 may be about 10mm and 20mm, respectively, wherein the dimension in the thickness direction is the arrangement direction of the heating body 23 and the accommodating chamber 21. As an example, the housing chamber 21 may also have a cylindrical structure with a diameter to thickness ratio greater than 5:1, for example, 0.3-1.0mm in thickness and 5-20mm in diameter. Of course, the cross section of the accommodating chamber 21 may be polygonal or elliptical, etc.
Because the accommodating cavity 21 is of a flat structure, the reaction sample arranged in the accommodating cavity 21 can be thin, the distance between the center of the reaction sample and the surface of the liquid is small, the temperature of the surface of the reaction sample can be quickly transferred to the center, the temperature of each part of the reaction sample can be consistent in a short time, the contact area between the reaction sample and the heating mechanism 2 is large due to the flat structure, the heat transfer efficiency is high, and the temperature rising and reducing speed and the detection efficiency of the reaction sample are greatly improved. The inner diameter of the PCR tube is larger than that of the flat accommodating cavity 21, the distance between the center of the reaction sample and the surface of the liquid is large, the temperature of the reaction sample needs a long time to be consistent, the temperature rising and reducing speed of the reaction sample is low, and the detection efficiency is low.
The accommodating cavity 21 and the heating mechanism 2 are of an integrated structure, the accommodating cavity 21 is directly formed on the heating mechanism 2, and the reaction sample is placed in the accommodating cavity 2, so that a heat transfer interface can be reduced, the heat conduction efficiency is improved, and the temperature rise and temperature reduction speed of the reaction sample is further improved. The nucleic acid amplification apparatus according to the present embodiment can realize 45 temperature cycles in about 15 minutes. When the nucleic acid amplification apparatus provided in the second embodiment performs the nucleic acid amplification method in the first embodiment, the reaction progress can be further accelerated.
As shown in fig. 5, at least part of the upper surface of the heating mechanism 2, as a cavity wall of the housing cavity 21, the housing cavity 21 may cover part of the upper surface of the heating mechanism 2 or entirely cover the upper surface of the heating mechanism 2. Or, as shown in fig. 6, at least part of the upper surface of the heating mechanism 2 at the partial depression is partially depressed as the cavity wall of the accommodating cavity 21, i.e., the upper surface of the heating mechanism 2, and the accommodating cavity 21 covers a part of the upper surface of the depression or completely covers the upper surface of the depression. The sample in the accommodating cavity 21 is directly contacted with the surface of the heating mechanism 2, so that a heat conduction interface is further reduced, the heat conduction efficiency is improved, and the temperature rising and reducing speed of the reaction sample is improved.
In other alternative embodiments, as shown in fig. 8, the heating means 2 comprises a means body and a bottom wall 22 of the receiving chamber 21, i.e. a bottom wall 22 is provided between the reaction sample and the means body. The bottom wall 22 is in close contact with at least part of the upper surface of the mechanism body, and the bottom wall 22 may cover part of the upper surface of the mechanism body or completely cover the upper surface of the mechanism body and be in close contact with the upper surface. Or the bottom wall 22 is in close contact with, i.e., partially recessed from, the upper surface of the mechanism body, and the receiving cavity 21 covers a portion of, or entirely covers, the upper surface of the recess. Since the bottom wall 22 is in close contact with the mechanism body, the air layer is reduced between the bottom wall 22 and the mechanism body, and heat can be rapidly transferred.
In order to make the bottom wall 22 closely contact with the mechanism body, alternatively, the bottom wall 22 and the mechanism body may be formed by injection molding, or of course, the bottom wall 22 and the mechanism body may be closely contacted by other means, which will not be described herein.
Of course, in other alternative embodiments, in order to avoid the presence of an air layer between the bottom wall 22 and the mechanism body, a flexible heat-conducting member 29 may be provided between the bottom wall 22 and the mechanism body. The flexible heat conducting member 29 can ensure a more conforming bond with the bottom wall 22 and the mechanism body, so that the bottom wall 22 and the mechanism body are both in close contact with the flexible heat conducting member 29, thereby avoiding the occurrence of an air layer. The flexible heat conductive member 29 may be a heat conductive silicone or the like.
Optionally, at least two independent subchambers are arranged in the accommodating chamber 21, and each subchamber can carry out a group of amplification reactions, so that the experimental efficiency is improved; or the number of the accommodating chambers 21 is one or at least two, and each accommodating chamber 21 can perform a set of amplification reactions, thereby improving experimental efficiency.
Alternatively, the heating mechanism 2 includes a heating body 23, the heating body 23 may be a copper wire or the like, and the temperature rise of the heating mechanism 2 is achieved by the heating body 23.
As shown in fig. 5, the heating mechanism 2 may further include an upper conductive member 24 and a lower conductive member 27, with the heating body 23 sandwiched between the upper conductive member 24 and the lower conductive member 27. The upper and lower conductive assemblies 24 and 27 have a conductive heat and insulating effect.
As shown in fig. 5, the upper conductive member 24 of the heating mechanism 2 may include a soaking layer 241, and the soaking layer 241 is in direct contact with the reaction sample in the receiving chamber 21. The soaking layer 241 can improve the uniformity of the temperature of the reaction sample and avoid the phenomenon that the temperature of a certain point is too high or too low. The soaking layer 241 can be made of metal such as aluminum or high heat conduction ceramic, so as to ensure heat conduction in the longitudinal direction and the transverse direction and ensure temperature uniformity of reaction sample liquid. The soaking layer 241 is in direct contact with the reaction sample in the accommodating cavity, so that a heat conducting layer can be reduced, and the heat conducting speed of the reaction sample is further improved.
As shown in fig. 5, the lower conductive assembly 27 may include an insulating thermal resistance layer 271. The insulating thermal resistance layer 271 has a certain thermal resistance characteristic and insulating characteristic. The insulating thermal resistance layer 271 may form a longitudinal thermal resistance in addition to insulating the heating body 23. The magnitude of the thermal resistance can be designed by material selection and thickness selection. Typically, the thermal resistance of this layer is much greater than that of the other layers of the structure, so that the insulating thermal resistance layer 271 is the main thermal resistance source for the heating mechanism 2 to dissipate heat and cool down to the cooling mechanism 1. The insulating thermal resistance layer 271 is one of the main factors that affect the thermal performance of the heating mechanism 2.
As shown in fig. 8, optionally, the cooling medium of the cooling mechanism 1 is directly contacted with the heating mechanism 2 to perform cooling, for example, the cooling mechanism 1 and the heating mechanism 2 are integrated, and the cooling mechanism 1 and the heating mechanism 2 share a heat conducting plate 12, so that one heat conducting surface is omitted, and the heating and cooling speeds are further improved.
In another alternative embodiment, shown in fig. 9, the cooling mechanism 1 includes a cooling body 13 and a cold head 14 connected to the cooling body 13, the heating mechanism 2 can be abutted against the cold head 14, and the cooling body 13 can cool the cold head 14. It should be noted that, the upper wall of the cooling body 13 may be formed as a cold head 14 (in fig. 7b, the cold head 14 is the heat-conducting plate 12), and as shown in fig. 9, the cold head 14 may be a block structure connected to the cooling body 13.
Alternatively, the cooling body 13 cools the cold head 14 by spraying the cooling medium, or the cooling medium that cools the cold head 14 flows through the cooling body 13. Preferably, the coldhead 14 is in direct contact with the cooling medium to reduce the heat transfer interface.
When the cooling medium flows through the cooling body 13, the cooling flow passage 11 may be provided in the cooling body 13. The cooling medium can flow through the cooling flow passage 11, so that the cooling mechanism 1 continuously cools the heating mechanism 2. The cooling medium may be water or other liquid.
Alternatively, as shown in fig. 3, the cooling body 13 may be grooved directly, the grooves forming the cooling flow passages 11 described above. For example, the cooling body 13 may include a first plate and a second plate that are sealed and fastened, and the first plate and/or the second plate are provided with grooves, and the first plate and the second plate are fastened and form a cooling flow channel.
Of course, in an alternative embodiment, the cooling body 13 may be provided with a cooling tube, which may be a pipe with good heat conduction, such as a copper pipe, and the space in the cooling tube forms the cooling flow channel 11. The cooling pipes can be arranged in a serpentine shape, and can be arranged in other shapes, so that the travel of cooling liquid in the cooling pipes is increased, and the cooling efficiency is improved.
As shown in fig. 6, the cooling mechanism 1 is not limited to the above-described cooling method, and the cooling mechanism 1 may be configured to perform cooling by continuously spraying a cooling medium to the heating mechanism 2. Alternatively, the cooling medium may be water or gas or the like (the arrows in fig. 6 indicate the partial flow direction of the cooling medium). For example, the cooling mechanism 1 may include a pump and a spray assembly in communication with the pump, the pump pumping a high pressure cooling medium into the spray assembly, the spray assembly spraying the cooling medium toward the heating mechanism 2.
As shown in fig. 5, to realize the temperature calibration function, the heating mechanism 2 may optionally further include a temperature calibration portion 25 for reflecting the temperature of the heating body 23.
The nucleic acid amplification apparatus may further include a temperature detection unit that detects the temperature of the temperature calibration section 25. The temperature of the heating body 23 can be more quickly represented to the temperature calibration part 25, that is, the temperature of the temperature calibration part 25 can be approximately equivalent to the temperature of the heating body 23 at the corresponding time, and thus, the temperature calibration part 25 can improve the detection efficiency of the temperature detection unit. The temperature detection unit may be any electronic device capable of detecting temperature, such as a temperature sensor.
However, in the prior art, since a certain time is required for transferring heat from the heating mechanism 2 to the temperature detecting unit of the sensor, the temperature detecting unit of the sensor may have a temperature measurement delay of 1-2s, and the temperature change of the heating mechanism 2 may reach more than 30 ℃ in the rapid temperature raising and lowering process for 1-2s, so that it is relatively difficult to control the heating mechanism 2 through the temperature detecting unit of the sensor in the rapid temperature raising and lowering process.
As shown in fig. 9, alternatively, the nucleic acid amplification apparatus may further include a resistance detection member 5 that obtains the temperature of the heating body 23 by resistance detection of the heating body 23. When measuring the temperature by the resistance temperature measurement method, firstly, the resistance value R of the heating body 23 is obtained by the resistance detection piece 5, and then the formula r=r is used 0 The temperature value of the heating body 23 at this time is obtained by the resistance value (1+αΔt), and then the temperature curve of the heating body 23 can be obtained by the continuous detection by the resistance detecting member 5.
The temperature value calibrated by the double temperature measurement mode is the temperature value obtained by the resistance detection piece 5. Due to nominal resistance R between resistors for the same type of resistor, e.g. copper wire resistor 0 And a slight difference in the temperature coefficient of resistance α, which causes a difference in the temperature measured by the resistance thermometry from the actual temperature. Whereas the temperature calibration value of the heating means 2 measured by the temperature detection unit is equal to the nominal resistance value R 0 And the temperature value obtained through the resistance detecting member 5 can be made more accurate after the calibration of the temperature coefficient of resistance alpha.
Specifically, the resistance detecting element 5 includes a first portion for detecting the voltage of the heating body 23 and a second portion for detecting the current of the heating body 23, and obtains the resistance of the heating body 23 from the voltage and the current, and further obtains the temperature value of the heating body 23 from the linear relationship between the resistance of the heating body 23 and the temperature. Since there is no delay in the voltage and current obtained by the resistance detecting member 5, the real-time temperature of the heating body 23 can be obtained by the resistance detecting member 5, and the accuracy of temperature control of the reaction sample can be improved.
The temperature detection method of the embodiment does not completely depend on the temperature value measured by an uncalibrated resistance temperature measurement method, and also does not completely depend on the temperature control heating mechanism 2 detected by the temperature detection unit, but the temperature detection unit and the resistance detection piece 5 are combined to jointly realize temperature detection, so that the problems of temperature detection delay and large temperature measurement error caused by a common temperature detection method in the prior art are overcome.
The temperature detection means may detect the temperature of the temperature calibration portion 25 in a noncontact or noncontact manner. As shown in fig. 5, 10, 12 and 14, when the temperature of the temperature calibration part 25 is detected in a contact or non-contact manner, for example, when the temperature detection unit is a contact type temperature detection unit or a non-contact type temperature detection unit 6, the temperature calibration part 25 is connected to the upper or lower conductive member 24 or 27 so that the temperature of the heating body 23 is conducted to the temperature calibration part 25. As shown in fig. 15, when the temperature detecting means is the noncontact temperature detecting means 6, the number of calibration portions may be one or two, as long as the temperature of the temperature calibration portion 25 can be detected by the noncontact temperature detecting means 6. As shown in fig. 5 and 10, alternatively, the two first contacts of the contact temperature detecting unit are respectively contacted with the two temperature calibrating parts 25, and no conduction exists between the two temperature calibrating parts 25, at this time, as shown in fig. 10, alternatively, the nucleic acid amplifying device may further include an external electrical connecting contact 251 and an electrical connecting lead 252, the number of the external electrical connecting contact 251 and the electrical connecting lead 252 may be two, the two external electrical connecting contacts 251 are respectively located at the sides of the two temperature calibrating parts 25 away from each other, one external connecting contact is electrically connected with one temperature calibrating part 25 through one electrical connecting lead 252, and the other external connecting contact is electrically connected with the other temperature calibrating part 25 through the other electrical connecting lead 252.
Referring to fig. 5 and 10, heat of the upper conductive member 24 (e.g., the soaking layer 241 of the upper conductive member 24 contacting the patch 261) is conducted to the temperature calibration part 25 through the patch 261 and the guide post 262, the temperature calibration part 25 is electrically connected to the outside through the electrical connection lead 252 at the external electrical connection contact 251, wherein the diameter of the electrical connection lead 252 is smaller than that of the temperature calibration part 25 and the external electrical connection contact 251, thereby reducing heat loss generated by the temperature calibration part 25 through the electrical connection lead 252, so that the temperature calibration part 25 can better reflect the temperature of the upper conductive member 24 (e.g., the soaking layer 241 of the upper conductive member 24 contacting the patch 261), the temperature detection unit realizes good electrical and thermal contact with the temperature calibration part 25 through the welding spot, and when the temperature of the upper conductive member 24 (e.g., the soaking layer 241 of the upper conductive member 24 contacting the patch 261) is changed, the temperature detection unit can quickly and accurately sense the temperature change, the temperature change is caused by the resistance change of the temperature detection unit, and the real-time temperature detection can be realized by detecting the resistance change of the temperature detection unit at the external electrical connection contact 251.
As shown in fig. 5, alternatively, the temperature calibration part 25 is located at the bottom of the heating mechanism 2. Alternatively, the bottom of the heating means 2 is the side of the heating means 2 remote from the receiving chamber 21. The position of the temperature calibration part 25 may be such that it is convenient to arrange the temperature detection unit.
As shown in fig. 9, in the present embodiment, the bottom of the heating mechanism 2 may be brought into contact with the cooling mechanism 1 to cool down the heating mechanism 2. In order to avoid the influence of the cooling mechanism 1 on the temperature of the temperature calibration portion 25, the cooling mechanism 1 is provided with an avoidance portion 15 for avoiding the temperature calibration portion 25 (in fig. 9, the temperature calibration portion 25 is a position facing the temperature detection means (specifically, the second contact temperature detection means 4 in fig. 9)), and the avoidance portion 15 may be a groove, a hole, or the like. Meanwhile, a temperature detecting unit (specifically, the second contact temperature detecting unit 4 in fig. 9) may be also provided at the avoiding portion 15 to make full use of the space and ensure that the temperature detecting unit can accurately detect the temperature of the temperature calibrating portion 25.
In connection with fig. 10 and 11, further, a receiving groove 275 is formed in a side of the lower conductive member 27 away from the heating body 23, and the temperature calibrating portion 25 is positioned in the receiving groove 275 and connected to a bottom of the receiving groove 275. Optionally, the accommodating groove 275 does not penetrate the lower conductive component 27, the bottom of the accommodating groove 275 is an insulating thermal resistance layer 271, and the temperature calibration part 25 is connected to the insulating thermal resistance layer 271. The receiving groove 275 may prevent the temperature calibrating portion 25 from protruding from the lower conductive member 27, thereby maintaining flatness of the lower surface of the heating mechanism 2 and facilitating smooth placement of the heating mechanism 2. Of course, in still another embodiment, the lower conductive member 27 may not be provided with the receiving groove 275, and the temperature calibrating portion 25 is connected to the lower surface of the lower conductive member 27.
In yet another alternative embodiment, as shown in fig. 12, the temperature calibration part 25 is connected to a side of the upper conductive assembly 24 adjacent to the lower conductive assembly 27, and a side of the upper conductive assembly 24 adjacent to the lower conductive assembly 27 is adjacent to the heating body 23, so that the temperature of the heating body 23 is rapidly transferred to the temperature calibration part 25.
The lower conductive member 27 may further be provided with a first through hole 273 disposed opposite to the temperature calibration portion 25 so that the temperature detecting member can detect the temperature of the temperature calibration portion 25. The lower surface of the upper conductive member 24 and the upper surface of the lower conductive member 27 are closest to the heating body 23, and the temperature thereof is closest to the temperature of the heating body 23 first, so that the temperature closest to the heating body 23 can be detected more quickly by the temperature detecting unit by connecting the temperature calibrating portion 25 to the side of the upper conductive member 24 closest to the lower conductive member 27.
As shown in fig. 5, alternatively, the temperature detecting unit may be the first contact temperature detecting unit 3, and the heating mechanism 2 may include the first contact temperature detecting unit 3 connected to the temperature calibrating portion 25 and configured to measure the temperature at the temperature calibrating portion 25. The first contact temperature detecting unit 3 may be a temperature sensor or the like. The first contact temperature detecting unit 3 may be connected to the temperature calibrating portion 25 by welding or the like, and after the heating mechanism 2 is used, the first contact temperature detecting unit 3 may be discarded together with the heating mechanism 2. Specifically, when the heating mechanism 2 is placed on the cooling mechanism 1, the first contact temperature detecting unit 3 may be provided at the escape portion 15 to make full use of space, and the temperature calibrating portion 25 may be directly opposite to the escape portion 15 to ensure consistency of the temperature at the temperature calibrating portion 25 and the temperature of the heating body 23.
As shown in fig. 9, the temperature detection unit in the present embodiment may be not the first contact temperature detection unit 3 but the second contact temperature detection unit 4. Specifically, the nucleic acid amplification apparatus includes the second contact temperature detection unit 4, and the second contact temperature detection unit 4 is capable of being separated from or in contact with the temperature calibration part 25, and is capable of measuring the temperature of the temperature calibration part 25 when it is in contact with the temperature calibration part 25.
Specifically, the second contact type temperature detecting unit 4 may be provided at the escape portion 15 to make full use of space and ensure that the temperature detecting unit can be brought into contact with the temperature calibrating portion 25. The temperature calibration portion 25 may be directly opposite to the avoidance portion 15 to ensure consistency of the temperature at the temperature calibration portion 25 with the temperature of the heating body 23. The second contact temperature detecting unit 4 is in elastic contact with the temperature calibrating portion 25, for example, the second contact temperature detecting unit 4 is connected with the cooling mechanism 1 through a spring, so as to achieve elastic contact. The second contact temperature detecting unit 4 is not discarded with the heating mechanism 2, and the cost of the heating mechanism 2 and the detecting cost can be reduced.
When the temperature of the temperature calibration portion 25 is detected in a non-contact manner, for example, when the temperature detection unit is a non-contact temperature detection unit 6 such as an infrared temperature measurement unit, the temperature calibration portion 25 may be provided in a manner other than the above-mentioned manner, as shown in fig. 13, and the temperature calibration portion 25 may be a part of the upper conductive member 24 or the lower conductive member 27, so that no additional connection of the temperature calibration portion 25 is required, and only the position of the temperature calibration portion 25 needs to be reserved for the upper conductive member 24 or the lower conductive member 27, so that the non-contact temperature detection unit 6 can be aligned with the position and the temperature of the position can be detected. Preferably, the lower conductive member 27 is provided with a second through hole 274 in the thickness direction of the heating mechanism 2, and the surface of the upper conductive member 24 facing the second through hole 274 is the temperature calibration portion 25. The lower surface of the upper conductive member 24 and the upper surface of the lower conductive member 27 are closest to the heating body 23, and the temperature thereof is closest to the temperature of the heating body 23 first, so that the temperature detection unit can more quickly detect the temperature closest to the heating body 23 by detecting the temperature of the lower surface of the upper conductive member 24.
Of course, in other alternative embodiments, the temperature calibration portion 25 may be a lower surface of the lower conductive member 27 to simplify the structure of the heating mechanism 2.
As shown in fig. 5, in order to shorten the time when the temperature of the temperature calibration portion 25 coincides with the temperature of the heating body 23, the heating mechanism 2 may optionally further include a rapid conduction portion 26, the rapid conduction portion 26 being configured to conduct heat of the heating body 23 to the temperature calibration portion 25. Specifically, in the present embodiment, the heat of the heating body 23 is indirectly transferred to the temperature calibration portion 25, for example, the heating member 23 heats the soaking layer 241, and the heat of the soaking layer 241 is transferred to the temperature calibration portion 25 through the rapid conduction portion 26, so that the temperature calibration portion 25 accurately reflects the temperature of the soaking layer 241, and the temperature detection unit can accurately measure the temperature of the soaking layer 241. Since the reaction sample has a small thickness, the temperature of the reaction sample substantially coincides with the temperature of the soaking layer 241, and the temperature of the reaction sample can be obtained by detecting the temperature of the temperature calibration section 25.
Preferably, one side of the rapid conduction part 26 is connected to one side of the upper conduction assembly 24 near the heating body 23, and the other side is connected to the temperature calibration part 25. The lower surface of the upper conductive member 24 and the upper surface of the lower conductive member 27 are closest to the heating body 23, and the temperature thereof is closest to the temperature of the heating body 23 first, so that the rapid conduction portion 26 is arranged in such a manner that the temperature of the rapid conduction portion 26 and the temperature of the heating body 23 are consistent in the shortest time. Alternatively, the rapid conduction portion 26 is made of a material having a high thermal conductivity, such as a metal material of copper or aluminum, or a thermally conductive ceramic, or the like. The thermal conductivity of the fast conducting portion 26 is particularly superior to that of the lower conducting assembly 27 to rapidly transfer heat to the temperature calibrating portion 25.
The quick-speed conduction part 26 comprises a patch 261 and one or more second guide posts 262, wherein the patch 261 is attached to one side of the upper conduction assembly 24 close to the heating body 23 or one side of the lower conduction assembly 27 close to the heating body 23, one end of the one or more second guide posts 262 is connected to the patch 261, and the other end of the one or more second guide posts 262 penetrates through the lower conduction assembly 27 and is connected with the temperature calibration part 25. The lower surface of the upper conductive member 24 and the upper surface of the lower conductive member 27 are closest to the heating body 23, and the temperature thereof is closest to the temperature of the heating body 23 first, so that the patch 261 is provided in such a manner that the temperature of the rapid conduction portion 26 is most rapidly coincident with the temperature of the heating body 23. The patch 261 can increase the contact area of the rapid conduction portion 26 with the upper conduction member 24 or the lower conduction member 27, improving the conduction efficiency. The cross-sectional area of the second guide post 262 can be smaller than that of the patch 261, so that the temperature of the patch 261 can be quickly conducted to the temperature calibration part 25, and meanwhile, the volume of the second guide post 262 can be reduced as much as possible, so that the influence of the quick conduction part 26 on the lower conduction assembly 27 is reduced, and the thermal resistance required by the thermal edge resistance layer 271 is ensured to be generated according to design. Alternatively, the patch 261 and the second guide post 262 are made of a material with high thermal conductivity such as copper, and when the patch 261 and the second guide post 262 are required to be insulating materials to avoid the heating mechanism 2 from being shorted, the patch 261 or the second guide post 262 may be made of a material with high thermal conductivity such as ceramic.
It is understood that the temperature calibration portions 25 may be disposed in one-to-one correspondence with the patches 261, and that two temperature calibration portions 25 may be connected to one patch 261. One temperature calibration part 25 may be connected to one second guide post 262, and in order to improve temperature uniformity of the temperature calibration part 25, the temperature calibration part 25 may be connected to a plurality of second guide posts 262.
As shown in fig. 14, in other alternative embodiments, the fast conducting portion 26 may include one or more first guide posts 263, where the fast conducting portion 26 is not provided with a patch 261, one end of the one or more first guide posts 263 is attached to a side of the upper conducting assembly 24 near the heating body 23, and the other end of the one or more first guide posts 263 is connected to the temperature calibration portion 25. The first guide post 263 does not affect other structures of the heating mechanism 2, and can rapidly transfer heat to the temperature calibration portion 25. Optionally, the first guide post 263 is a metal post with high thermal conductivity such as a copper post. One temperature calibration part 25 may be connected to one first guide post 263, and in order to improve temperature uniformity of the temperature calibration part 25, the temperature calibration part 25 may be connected to a plurality of first guide posts 263.
Optionally, the lower conductive component 27 is provided with a third through hole, and the first guide post 263 or the second guide post 262 is disposed in the third through hole. The third through hole can facilitate the placement of the first guide post 263 or the second guide post 262, and meanwhile, the continuity of the lower conductive assembly 27 is not affected, and the performance of the lower conductive assembly 27 is ensured. Optionally, the first guide post 263 or the second guide post 262 is not in contact with the inner wall of the third through hole, so as to avoid heat conduction between the first guide post 263 or the second guide post 262 and the lower conductive component 27, thereby affecting the temperature of the temperature calibration portion 25.
As shown in fig. 11, in order to obtain the resistance of the heating body 23 and supply power to the heating body 23, optionally, a plurality of second contacts 20 are disposed on the side of the heating mechanism 2 away from the accommodating cavity 21, the second contacts 20 are electrically connected with the heating body 23, and the second contacts 20 are also electrically connected with the resistance detecting member 5. The current and voltage of the heating body 23 can be obtained through the second contact 20, and thus the resistance value of the heating body 23 can be obtained.
In this embodiment, the second contact 20 enables the heating mechanism 2 to achieve its own temperature measurement function, compared with the conventional structure that can only measure the temperature through the external temperature measurement unit, the embodiment can directly measure the temperature of the heating mechanism 2 itself, so that the temperature measurement is more accurate and rapid, and the accuracy and control speed of the temperature control system can be improved.
The temperature detection unit may also be a non-contact temperature detection unit 6 for measuring the temperature at the temperature calibration part 25.
Specifically, the noncontact temperature detecting unit 6 may be provided at the escape portion 15 to make full use of space and to ensure that the noncontact temperature detecting unit 6 detects the temperature of the temperature calibrating portion 25. The non-contact temperature detection unit 6 is not discarded with the heating mechanism 2, and the cost of the heating mechanism 2 and the detection cost can be reduced.
In the prior art, the temperature of the heating mechanism 2 is detected by the isothermal detection unit of the temperature sensor, but since a certain time is required for heat transfer from the heating mechanism 2 to the isothermal detection unit, a temperature measurement delay of 1-2s exists in a detection result measured by the isothermal detection unit under normal conditions, and the temperature change of the heating mechanism 2 can reach more than 30 ℃ in a rapid temperature rise and fall process, so that the control of the heating mechanism 2 by the isothermal detection unit is relatively difficult in the rapid temperature rise and fall process. The temperature of the heating mechanism 2 is not completely dependent on the temperature value measured by an uncalibrated resistance temperature measuring method, and the heating mechanism 2 is not completely dependent on the temperature detected by the temperature detecting unit, but the temperature of the heating mechanism 2 is measured by adopting the temperature measuring part 25 of the heating mechanism 2 and the temperature of the heating body 23 is measured by using the resistance temperature measuring method, so that the temperature of the heating mechanism 2 can be rapidly and accurately controlled, the aim of accurately controlling the temperature is fulfilled, and the problems of temperature detection delay and large temperature measurement error caused by a common temperature detecting method in the prior art are solved.
Optionally, the nucleic acid amplification apparatus further comprises a temperature control unit for controlling the temperature of the reaction sample. Specifically, the temperature control unit is electrically connected to the resistance detecting member 5, and is capable of converting data acquired by the first and second portions of the resistance detecting member 5 into the temperature of the heating body 23, and the temperature of the reaction sample can be controlled by controlling the heating mechanism 2. For example, the temperature control unit controls the current, voltage and/or power of the heating mechanism 2 through an algorithm, and further controls the temperature of the heating mechanism 2. The control method reduces delay and has high control precision. Further, for example, when the temperature is higher than a preset temperature, the temperature control unit controls the heating mechanism 2 to reduce power or turns off the heating mechanism 2; when the temperature is lower than the preset temperature, the temperature control unit controls the heating mechanism 2 to increase the power so as to be maintained in the preset temperature. In the heating process, the temperature control unit achieves the purpose of heating by increasing the power of the heating mechanism 2, and in the cooling process, the temperature control unit achieves the purpose of cooling by reducing the power of the heating mechanism 2. In the heat preservation and temperature rising process, the temperature of the cooling mechanism 1 can be controlled by the control unit to assist in achieving temperature rising and heat preservation so as to reduce the energy consumption of the heating mechanism part 2.
In this embodiment, the temperature control unit may be a centralized or distributed controller, for example, the controller may be a single-chip microcomputer, or may be a distributed multi-chip microcomputer, where a control program may be run in the single-chip microcomputer, so as to control the above components to implement functions thereof.
The working procedure of the nucleic acid amplification apparatus according to this embodiment is as follows:
injecting a reaction sample into the accommodating chamber 21;
and starting the cooling mechanism 1, wherein the cooling mechanism 1 keeps continuously refrigerating the reaction sample, and the temperature control unit controls the heating mechanism 2 to work until the nucleic acid amplification is completed.
While the utility model has been described in detail in the foregoing general description, embodiments and experiments, it will be apparent to those skilled in the art that modifications and improvements can be made thereto. Accordingly, such modifications or improvements may be made without departing from the spirit of the utility model and are intended to be within the scope of the utility model as claimed.
Claims (21)
1. The nucleic acid amplification device is characterized by comprising a cooling mechanism and a heating mechanism, wherein the heating mechanism is provided with a containing cavity, the containing cavity is of a flat structure, and the surface, close to the cooling mechanism, of the heating mechanism is formed by a heat conducting material.
2. The nucleic acid amplification apparatus according to claim 1, wherein the cooling medium of the cooling mechanism is cooled in direct contact with the heating mechanism.
3. The nucleic acid amplification apparatus of claim 1, wherein at least part of an upper surface of the heating mechanism or at least part of an upper surface of a recess of the heating mechanism portion serves as a bottom wall of the accommodating chamber.
4. The nucleic acid amplification apparatus of claim 3, wherein the upper surface of the heating mechanism comprises a soaking layer that is in direct contact with the reaction sample in the receiving chamber.
5. The nucleic acid amplification apparatus according to claim 3, wherein the heating mechanism includes a mechanism body and a bottom wall of the accommodation chamber;
the bottom wall is closely contacted with at least part of the upper surface of the mechanism body, or the bottom wall is closely contacted with the upper surface of the recess of the mechanism body, or
A flexible heat conducting element is arranged between the bottom wall and the mechanism body.
6. The nucleic acid amplification apparatus of claim 1, wherein at least two independent subchambers are provided within the receiving chamber; or the number of the accommodating cavities is one or at least two.
7. The nucleic acid amplification apparatus according to claim 1, wherein the heating mechanism includes a heating body and a temperature calibration section for reflecting a temperature of the heating body.
8. The nucleic acid amplification apparatus of claim 7, further comprising a resistance detection member for detecting a resistance of the heating body.
9. The nucleic acid amplification apparatus of claim 8, wherein the heating mechanism further comprises an upper conductive member and a lower conductive member, and the heating body is sandwiched between the upper conductive member and the lower conductive member.
10. The nucleic acid amplification apparatus according to claim 9, wherein the heating mechanism further comprises a rapid conduction portion for conducting heat of the heating body to the temperature calibration portion.
11. The nucleic acid amplification apparatus of claim 10, wherein one side of the rapid conduction portion is connected to one side of the upper conduction assembly close to the heating body or to one side of the lower conduction assembly close to the heating body, and the other side is connected to the temperature calibration portion.
12. The nucleic acid amplification apparatus of claim 11, wherein the temperature calibration portion is located at a bottom of the heating mechanism.
13. The nucleic acid amplification apparatus according to claim 11, wherein a receiving groove is provided on a side of the lower conduction assembly away from the heating body, and the temperature calibration part is located in the receiving groove and connected to a bottom of the receiving groove.
14. The nucleic acid amplification apparatus of claim 11, wherein the rapid conduction portion comprises one or more first guide posts, one end of the one or more first guide posts is attached to a side of the upper conduction assembly close to the heating body or to a side of the lower conduction assembly close to the heating body, and the other end of the one or more first guide posts is connected to the temperature calibration portion.
15. The nucleic acid amplification apparatus of claim 11, wherein the rapid conduction portion comprises a patch and one or more second guide posts, the patch is attached to a side of the upper conduction assembly close to the heating body or attached to a side of the lower conduction assembly close to the heating body, one end of the one or more second guide posts is connected to the patch, and the other end of the one or more second guide posts penetrates through the lower conduction assembly and is connected to the temperature calibration portion.
16. The nucleic acid amplification apparatus of claim 15, wherein the lower conductive element is provided with a third through hole, and the second guide post is disposed in the third through hole.
17. The nucleic acid amplification apparatus according to claim 8, further comprising a temperature detection unit for detecting a temperature of the temperature calibration part.
18. The nucleic acid amplification apparatus according to claim 17, wherein the cooling mechanism is provided with a avoiding portion for avoiding the temperature calibration portion and the temperature detection unit.
19. The nucleic acid amplification apparatus according to claim 1, wherein the cooling mechanism comprises a cooling body and a cold head connected to the cooling body, the heating mechanism is capable of being brought into close contact with the cold head, and the cooling body is capable of cooling the cold head.
20. The nucleic acid amplification apparatus of claim 19, wherein the cooling body cools the coldhead by spraying a cooling medium; or alternatively, the first and second heat exchangers may be,
the cooling medium for cooling the cold head flows through the cooling main body.
21. The nucleic acid amplification apparatus of claim 1, further comprising a temperature control unit.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202320882154.2U CN219792989U (en) | 2023-04-18 | 2023-04-18 | Nucleic acid amplification device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202320882154.2U CN219792989U (en) | 2023-04-18 | 2023-04-18 | Nucleic acid amplification device |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN219792989U true CN219792989U (en) | 2023-10-03 |
Family
ID=88184773
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202320882154.2U Active CN219792989U (en) | 2023-04-18 | 2023-04-18 | Nucleic acid amplification device |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN219792989U (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117327571A (en) * | 2023-11-30 | 2024-01-02 | 广州国家实验室 | Carrier and application thereof |
-
2023
- 2023-04-18 CN CN202320882154.2U patent/CN219792989U/en active Active
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117327571A (en) * | 2023-11-30 | 2024-01-02 | 广州国家实验室 | Carrier and application thereof |
| CN117327571B (en) * | 2023-11-30 | 2024-05-14 | 广州国家实验室 | Carrier and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP5996613B2 (en) | Improvement of thermocycler for PCR | |
| CN219792989U (en) | Nucleic acid amplification device | |
| JP4579993B2 (en) | Differential scanning calorimeter (DSC) with temperature controlled furnace | |
| US8151575B2 (en) | Temperature variation apparatus | |
| US20070230535A1 (en) | Thermal cycler for PCR | |
| CN103260538B (en) | Therapeutic apparatus and control method therefor | |
| CN100595502C (en) | Defrosting apparatus of refrigerator | |
| KR102204931B1 (en) | Thermal cycler sample block with function of selective temperature control for PCR | |
| US20060081598A1 (en) | Portable blackbody furnace | |
| CN118813769A (en) | Nucleic acid amplification method and nucleic acid amplification device | |
| WO2024217420A1 (en) | Nucleic acid amplification method and nucleic acid amplification apparatus | |
| CN219792943U (en) | Reaction module and nucleic acid amplification device | |
| CN108931087A (en) | A kind of refrigerating appliance | |
| CN101735949B (en) | Temperature change device | |
| CN111092362B (en) | Laser frequency stabilization method and device based on temperature self-sensing flexible thin film heater | |
| CN118813376A (en) | Reaction module, nucleic acid amplification device and nucleic acid amplification control method | |
| WO2024217049A1 (en) | Reaction assembly, nucleic acid amplification device, and nucleic acid amplification control method | |
| CN220788608U (en) | Nucleic acid amplification device and nucleic acid detection system | |
| CN111064070B (en) | Laser frequency stabilization method and device based on laser tube surrounding type temperature measurement and accurate temperature control point | |
| KR20100117922A (en) | Pcr apparatus increasing dna | |
| CN211329442U (en) | Test card incubator structure | |
| WO2024217421A1 (en) | Nucleic acid amplification device and nucleic acid test system | |
| CN118813399A (en) | Nucleic acid amplification device and nucleic acid amplification system | |
| CN109827570A (en) | Inertia measuring module | |
| CN215679153U (en) | Temperature testing device |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| GR01 | Patent grant | ||
| GR01 | Patent grant |