A reaction assembly, a nucleic acid amplification device, and a nucleic acid amplification control method

Abstract

The present application relates to a kind of reaction components, nucleic acid amplification device and nucleic acid amplification control method.Reaction components, including for accommodating reaction sample accommodating cavity and the heater of heating to the reaction sample.Nucleic acid amplification device includes reaction components.Nucleic acid amplification control method includes: by resistance thermometry and the temperature of the reaction component is calibrated double thermometry mode control the reaction component.Heater and accommodating cavity are integrated structure, heater and accommodating cavity are connected closely, there is no air layer, so as to accelerate the heat transfer speed of heater and the reaction sample in accommodating cavity, in turn, accelerate nucleic acid amplification process, improve detection efficiency.Nucleic acid amplification control method can realize the temperature of fast and accurately control reaction component, reach the purpose of accurate temperature control.

Description

Technical Field

[0001] This invention relates to the field of in vitro diagnostic technology, and in particular to a reaction component, a nucleic acid amplification device, and a nucleic acid amplification control method. Background Technology

[0002] PCR (polymerase chain reaction) is a molecular biology experimental method for the in vitro enzymatic synthesis of specific DNA fragments. PCR amplification, i.e. nucleic acid amplification, mainly consists of repeated thermal cycles of three steps: high-temperature denaturation, low-temperature annealing, and appropriate-temperature extension.

[0003] In existing technologies, a housing or tube (such as a PCR tube) with a receiving cavity is placed on a heater to facilitate later removal of the housing or tube. The receiving cavity is used to hold the reaction sample, and the heater heats the housing or tube, transferring heat to the reaction sample to achieve amplification. However, because the housing or tube is only placed on the heater for heating, an air layer exists between the housing or tube and the heater, reducing the heat transfer rate between them. This results in slow heating and cooling of the reaction sample, thus affecting the nucleic acid amplification process and detection efficiency. Furthermore, the large thickness of the reaction sample inside the housing or tube leads to a longer time required for homogenization, also contributing to slow heating and cooling.

[0004] In the prior art, a multi-layer structure is provided between the heater and the reaction sample in the containment cavity. The multi-layer structure further hinders the heat transfer between the heater and the reaction sample, which also results in a slow heating and cooling rate of the reaction sample.

[0005] To facilitate heater control, the temperature of the reaction sample needs to be detected. Currently, there are two main methods for temperature detection. The first method uses a temperature sensor to detect either the heater temperature or the reaction sample temperature. However, because heat transfer to the temperature sensor takes time, the sensor readings are delayed by 1-2 seconds, leading to inaccurate heater control and significantly affecting the results. The second method involves detecting the heater's resistance and then combining this with the temperature coefficient of resistance (TCR) and the nominal resistance value to arrive at the corresponding temperature. However, for resistors of the same type, such as copper wire resistors, slight differences in nominal resistance and TCR can result in large temperature measurement errors, further affecting the detection results. Summary of the Invention

[0006] One object of the present invention is to provide a reaction assembly to at least solve one of the above-mentioned problems.

[0007] To achieve the above objectives, a first aspect of the present invention provides a reaction assembly, including a receiving cavity for containing a reaction sample and a heater for heating the reaction sample.

[0008] Optionally, the heater is in direct contact with the reaction sample within the containment cavity.

[0009] Optionally, at least a portion of the upper surface of the heater is in direct contact with the reaction sample within the containment cavity, or

[0010] The surface of the heater is formed with grooves, and at least a portion of the wall of the grooves is in direct contact with the reaction sample inside the containment cavity.

[0011] Optionally, the heater includes a heat spreader layer that is in direct contact with the reaction sample within the containment cavity.

[0012] Optionally, the heater further includes a heating element and a temperature calibration unit for the temperature detection unit to detect the temperature.

[0013] Optionally, the heater further includes an upper conductive assembly and a lower conductive assembly, with the heating element sandwiched between the upper conductive assembly and the lower conductive assembly, and the upper conductive assembly including the heat spreader layer.

[0014] Optionally, the temperature calibration unit is connected to the upper conduction component or the lower conduction component.

[0015] Optionally, the temperature calibration unit is connected to the side of the upper conduction component near the lower conduction component, and the lower conduction component has a first through hole that is directly opposite the temperature calibration unit.

[0016] Optionally, the temperature calibration unit is part of the upper conduction component or the lower conduction component.

[0017] Optionally, the lower conductive component has a second through hole along the thickness direction of the reaction component, and the surface of the upper conductive component opposite to the second through hole is the temperature calibration part.

[0018] Optionally, the reaction assembly further includes a rapid conduction section for conducting heat from the heating element to the temperature calibration section.

[0019] Optionally, one side of the rapid conduction section is connected to the side of the upper conduction assembly near the heating element or to the side of the lower conduction assembly near the heating element, and the other side is connected to the temperature calibration section.

[0020] Optionally, the temperature calibration unit is located on the side of the lower conduction assembly away from the heating element.

[0021] Optionally, the lower conduction component has a receiving groove on the side away from the heating element, and the temperature calibration part is located in the receiving groove and connected to the bottom of the receiving groove.

[0022] Optionally, the rapid conduction section includes one or more first guide posts, one end of which is attached to the side of the upper conduction assembly near the heating element or to the side of the lower conduction assembly near the heating element, and the other end of which is connected to the temperature calibration section.

[0023] Optionally, the rapid conduction section includes a patch and one or more second guide posts. The patch is attached to the side of the upper conduction assembly near the heating element or to the side of the lower conduction assembly near the heating element. One end of the one or more second guide posts is connected to the patch, and the other end passes through the lower conduction assembly and is connected to the temperature calibration section.

[0024] Optionally, the lower conductive component has a third through hole, and the guide post is disposed in the third through hole.

[0025] Optionally, the lower conductive component further includes an insulating thermal resistance layer.

[0026] Optionally, the lower conductive component further includes a thermally conductive layer located on the side of the insulating thermal resistance layer away from the heating element.

[0027] Optionally, the heat spreader is made of a conductive or insulating material.

[0028] Optionally, when the heat spreader is made of a conductive material, the upper conductive assembly further includes an insulating layer located between the heating element and the heat spreader.

[0029] When the heat spreader is made of an insulating material, the heat spreader is adjacent to the heating element.

[0030] Optionally, the receiving cavity includes a bottom wall;

[0031] The bottom wall is in close contact with at least a portion of the upper surface of the heater, or

[0032] The surface of the heater is formed with a groove, and at least a portion of the wall of the groove is in close contact with the bottom wall.

[0033] Optionally, a flexible heat-conducting element is provided between the bottom wall and the heater.

[0034] Optionally, the receiving cavity has a flat structure.

[0035] Optionally, the cross-section of the receiving cavity is polygonal, circular, or elliptical.

[0036] Optionally, the reaction assembly includes a first contact temperature detection unit connected to the temperature calibration unit and used to measure the temperature at the temperature calibration unit.

[0037] Another object of the present invention is to provide a nucleic acid amplification device for performing nucleic acid amplification.

[0038] To achieve this objective, the second aspect of the present invention adopts the following technical solution:

[0039] A nucleic acid amplification device, comprising the aforementioned reaction components.

[0040] Another object of the present invention is to provide a nucleic acid amplification device for performing nucleic acid amplification.

[0041] To achieve this objective, the third aspect of the present invention adopts the following technical solution:

[0042] A nucleic acid amplification device,

[0043] It includes a second contact temperature detection unit and the reaction component. The second contact temperature detection unit is capable of separating from or contacting the temperature calibration part. When it is in contact with the temperature calibration part, it is capable of measuring the temperature of the temperature calibration part.

[0044] Another object of the present invention is to provide a nucleic acid amplification device for performing nucleic acid amplification.

[0045] To achieve this objective, the fourth aspect of the present invention adopts the following technical solution:

[0046] A nucleic acid amplification device includes a non-contact temperature detection unit and a reaction assembly as described above, wherein the non-contact unit is used to measure the temperature at the temperature calibration section.

[0047] Optionally, the nucleic acid amplification device further includes a cooling mechanism for cooling the reaction sample within the containment chamber.

[0048] Optionally, the cooling mechanism has a clearance portion on the side near the reaction component.

[0049] Optionally, the cooling mechanism cools the reaction components via fluid.

[0050] Optionally, the cooling mechanism cools the reaction components by injecting fluid or by fluid flow.

[0051] Optionally, when the cooling mechanism cools the reaction component by fluid flow, the cooling mechanism includes a cooling body with cooling channels provided inside to allow the cooling medium to flow.

[0052] Optionally, the cooling mechanism uses solid cooling to cool the reaction components.

[0053] Optionally, the nucleic acid amplification device further includes a resistance detection unit for detecting the temperature of the reaction components.

[0054] Optionally, the resistance detection unit obtains the temperature value of the reaction component by detecting the resistance of the reaction component.

[0055] Another object of the present invention is to provide a method for controlling nucleic acid amplification in order to perform nucleic acid amplification.

[0056] To achieve this objective, the fifth aspect of the present invention adopts the following technical solution:

[0057] A nucleic acid amplification control method, wherein the nucleic acid amplification control method is performed using the reaction components or nucleic acid amplification device described above;

[0058] The nucleic acid amplification control method includes controlling the reaction component using a dual temperature measurement method, namely, resistance thermometry and temperature calibration of the reaction component.

[0059] Optionally, controlling the reaction assembly using a dual temperature measurement method—resistance thermometry and temperature calibration of the reaction assembly—includes:

[0060] The temperature calibration value of the reaction component is measured by a temperature detection unit, and the temperature value obtained by the resistance value of the reaction component is calibrated by the temperature calibration value.

[0061] Optionally, the temperature coefficient of resistance and nominal resistance of the reaction component can be obtained based on the temperature calibration value to calibrate the temperature value.

[0062] Optionally, the temperature value can be calibrated by measuring the temperature calibration value before the nucleic acid amplification process, during the first temperature rise in the nucleic acid amplification process, and / or during the first amplification cycle in the nucleic acid amplification process.

[0063] Optionally, the temperature value can be calibrated using at least two unequal temperature calibration values.

[0064] Optionally, controlling the reaction assembly using a dual temperature measurement method—resistance thermometry and temperature calibration of the reaction assembly—includes the following steps:

[0065] At least two different temperature calibration values ​​are obtained; a first voltage and a first current of the reaction component at one temperature calibration value are detected, and a first resistance value of the reaction component is obtained based on the first voltage and the first current; a second voltage and a second current of the reaction component at another temperature calibration value are detected, and a second resistance value of the reaction component is obtained based on the second voltage and the second current;

[0066] Based at least on the first resistance value, the second resistance value, and their corresponding temperature calibration values, the resistance temperature coefficient and resistance at the nominal temperature of the reaction component can be obtained;

[0067] The current and voltage of the reaction component are continuously monitored, and the reaction component is controlled based on the temperature coefficient of resistance and the resistance at the nominal temperature.

[0068] Optionally, before calibrating the temperature value, the temperature of the reaction component is controlled by the temperature of the reaction component measured by the temperature detection unit, or the temperature of the reaction component is controlled according to a preset RT temperature curve of the reaction component.

[0069] Alternatively, according to the formula: R=R0(1+α) T) The temperature coefficient of resistance and nominal resistance of the reaction component are calibrated, and the temperature curve of the reaction component is obtained according to the formula by continuously measuring the voltage and current of the reaction component, where R0 is the nominal resistance and α is the temperature coefficient of resistance of the material.

[0070] As can be seen from the above, the technical solution provided by this invention includes a reaction assembly comprising a cavity for containing a reaction sample and a heater for heating the reaction sample. The heater and the cavity are an integral structure, tightly connected without an air layer, thereby accelerating the heat transfer rate between the heater and the reaction sample within the cavity, thus accelerating the nucleic acid amplification process and improving detection efficiency. The heater is in direct contact with the reaction sample within the cavity, and the cavity is directly formed on the upper surface of the heater. There are no other conductive interfaces between the reaction sample and the heater, thereby reducing the conductive interface between the heater and the cavity and further improving conductivity. Attached Figure Description

[0071] Figure 1 This is a schematic diagram of the structure of the first reaction component provided in an embodiment of the present invention;

[0072] Figure 2 This is a schematic diagram of the structure of the second reaction component provided in an embodiment of the present invention;

[0073] Figure 3 This is a schematic diagram of the structure of the third reaction component and the cooling mechanism for fluid flow provided in the embodiments of the present invention;

[0074] Figure 4 This is a schematic diagram of the structure of the fourth reaction component provided in the embodiments of the present invention;

[0075] Figure 5 This is a schematic diagram of the structure of the fifth reaction component provided in the embodiments of the present invention;

[0076] Figure 6 This is a schematic diagram of the structure of the sixth reaction component provided in this embodiment of the invention;

[0077] Figure 7This is a schematic diagram of the structure of the seventh reaction component provided in the embodiments of the present invention;

[0078] Figure 8 This is a schematic diagram of the structure of the eighth reaction component provided in this embodiment of the invention;

[0079] Figure 9 This is a schematic diagram of the structure of the ninth reaction component provided in the embodiments of the present invention;

[0080] Figure 10 This is a schematic diagram of the structure of the tenth reaction component provided in this embodiment of the invention;

[0081] Figure 11 This is a schematic diagram of the structure of the reaction assembly and the cooling mechanism of the jet fluid provided in an embodiment of the present invention;

[0082] Figure 12 This is a schematic diagram of the structure of the nucleic acid amplification device provided in this embodiment of the invention when the first contact temperature detection unit is used for temperature measurement;

[0083] Figure 13 This is a partial cross-sectional view of the nucleic acid amplification device provided in this embodiment of the invention when using a second contact temperature detection unit for temperature measurement;

[0084] Figure 14 This is a schematic diagram of the structure of the nucleic acid amplification device provided in this embodiment of the invention when the second contact temperature detection unit is used for temperature measurement;

[0085] Figure 15 This is a schematic diagram of the structure of the nucleic acid amplification device provided in this embodiment of the invention when using a non-contact temperature detection unit for temperature measurement;

[0086] Figure 16 This is a flowchart of the nucleic acid amplification control method provided in the embodiments of the present invention;

[0087] Figure 17 This is the temperature-time curve of the reaction component provided in the embodiment of the present invention.

[0088] In the picture:

[0089] 1. Cooling mechanism; 11. Cooling flow channel; 12. Cooling body; 13. Clearance section;

[0090] 2. Reaction components;

[0091] 21. Receiving cavity; 22. Bottom wall; 23. Heating element; 24. Upper conduction assembly; 241. Heat spreader layer; 242. Insulating layer; 25. Temperature calibration section; 251. External electrical contact point; 252. Electrical connection lead; 26. Rapid conduction section; 261. Patch; 262. Second guide post; 263. First guide post; 27. Lower conduction assembly; 271. Insulating thermal resistance layer; 272. Thermally conductive layer; 273. First through hole; 274. Second through hole; 275. Receiving groove; 28. Cavity sidewall; 29. ​​Cover plate; 20. Second contact point; 201. Groove; 202. Wall surface; 2021. Sidewall; 2022. Groove bottom; 203. Flexible thermally conductive element;

[0092] 200. Heater;

[0093] 3. First contact temperature detection unit; 4. Second contact temperature detection unit; 5. Non-contact temperature detection unit; 6. Resistance detection unit. Detailed Implementation

[0094] The technical solution of the present invention will be further described below with reference to the accompanying drawings and specific embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and not intended to limit it. Furthermore, it should be noted that, for ease of description, only the parts relevant to the present invention are shown in the accompanying drawings, not all of them.

[0095] This invention defines certain directional terms. Unless otherwise stated, the directional terms used, such as "up," "down," "left," "right," "inner," and "outer," are used for ease of understanding and therefore do not constitute a limitation on the scope of protection of this invention.

[0096] In this invention, unless otherwise explicitly specified and limited, "above" or "below" the second feature can include direct contact between the first and second features, or contact between the first and second features through another feature between them. Furthermore, "above," "over," and "on top" of the second feature includes the first feature directly above or diagonally above the second feature, or simply indicates that the first feature is at a higher horizontal level than the second feature. "Below," "below," and "under" the second feature includes the first feature directly below or diagonally below the second feature, or simply indicates that the first feature is at a lower horizontal level than the second feature.

[0097] In the description of this invention, unless otherwise explicitly specified and limited, the terms "connected," "linked," and "fixed" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral part; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; they can refer to the internal communication of two components or the interaction between two components. Those skilled in the art can understand the specific meaning of the above terms in this invention based on the specific circumstances.

[0098] Example 1

[0099] This embodiment provides a reaction component 2 for amplifying reaction samples, but it is not limited to this and can also be used in other situations where the reaction samples need to be heated to improve detection efficiency.

[0100] like Figures 1-10 As shown, the reaction assembly 2 provided in this embodiment includes a containment cavity 21 for containing reaction samples and a heater 200 for heating the reaction samples. The heater 200 and the containment cavity 21 are an integral structure, and the heater 200 and the containment cavity 21 are tightly connected without an air layer, thereby accelerating the heat transfer rate between the heater 200 and the reaction samples in the containment cavity 21, thereby accelerating the nucleic acid amplification process and improving detection efficiency.

[0101] like Figure 1 and Figure 2 As shown, the heater 200 is in direct contact with the reaction sample inside the receiving cavity 21. There are no other conductive interfaces between the reaction sample and the heater 200, thereby reducing the conductive interface between the heater 200 and the receiving cavity 21 and further improving the conduction efficiency. At the same time, the absence of an interface between the heater 200 and the reaction sample results in lower thermal resistance, enabling faster temperature control while achieving rapid heat conduction.

[0102] At least a portion of the upper surface of the heater 200 is in direct contact with the reaction sample in the containment cavity 21, that is, the containment cavity 21 is directly formed on the upper surface of the heater 200, and there are no other conductive interfaces between the reaction sample and the heater 200, thereby reducing the conductive interface between the heater 200 and the containment cavity 21 and further improving the conduction efficiency.

[0103] like Figure 1 As shown, optionally, an annular cavity sidewall 28 can be connected to the upper surface of the heater 200, with the top end of the cavity sidewall 28 connected to a cover plate 29. The cavity sidewall 28, the cover plate 29, and at least a portion of the upper surface of the heater 200 form the aforementioned receiving cavity 21, thereby containing and confining the reaction sample. The cover plate 29 can be made of glass or transparent plastic to allow optical detection of the sample liquid from above.

[0104] In one specific embodiment, the cover plate 29 is made of glass, with an exemplary thickness of 0.2 mm. The cavity sidewall 28 supports the cover plate 29, and, as an example, its height is 0.5 mm, meaning the thickness of the receiving cavity 21 is 0.5 mm. When the reaction sample fills the receiving cavity 21, the thickness of the reaction sample is 0.5 mm. The cover plate 29 has a pre-drilled injection hole for loading the reaction sample, which can be sealed with a film after the reaction sample is placed inside.

[0105] Optionally, the receiving cavity 21 has a flat structure. This means that the thickness dimension of the receiving cavity 21 is much smaller than its width or length dimension. For example, the receiving cavity 21 can be a cuboid, with a length-to-thickness ratio greater than 5:1, such as 90:1. The thickness dimension of the receiving cavity 21 can be 0.3-1.0 mm, and its width and length are approximately 10 mm and 20 mm, respectively. The thickness dimension refers to the arrangement direction of the heating element 23 and the receiving cavity 21. Alternatively, the receiving cavity 21 can also be a cylindrical structure with a diameter-to-thickness ratio greater than 5:1, such as a thickness of 0.3-1.0 mm and a diameter of 5-20 mm. Of course, the cross-section of the receiving cavity 21 can be polygonal or elliptical, etc.

[0106] The reaction sample in the flat-structured cavity 21 is thin, and the center of the reaction sample is very close to the liquid surface. This allows the temperature of the reaction sample to reach uniformity in a very short time. Furthermore, the flat structure provides a large contact area between the reaction sample and the heater 200, resulting in high heat transfer efficiency and significantly improved heating / cooling rates and detection efficiency. In contrast, the inner diameter of the PCR tube is relatively large compared to the flat-structured cavity 21. This results in a much greater distance between the center of the reaction sample and the liquid surface, requiring a longer time for the temperature of the reaction sample to reach uniformity. Consequently, the heating / cooling rates are slower, leading to lower detection efficiency.

[0107] like Figure 2 As shown, in other optional embodiments, a groove 201 is formed on the surface of the heater 200, and at least a portion of the wall 202 of the groove 201 is in direct contact with the reaction sample in the receiving cavity 21. That is, the reaction sample is in direct contact with the wall 202 of the groove 201, and there are no other conductive interfaces between the reaction sample and the heater 200, thereby reducing the conductive interface between the heater 200 and the receiving cavity 21 and further improving the conduction efficiency. At the same time, the sidewall 2021 of the wall 202 of the groove 201 is also in contact with the reaction sample. Compared with the heater 200 without the groove 201, the contact area between the heater 200 and the reaction sample can be increased, further increasing the heat transfer efficiency.

[0108] like Figure 2As shown, preferably, the receiving cavity 21 coincides with the groove 201, that is, the space inside the groove 201 forms the receiving cavity 21. The reaction sample directly contacts the side wall 2021 and the bottom 2022 of the wall 202 of the groove 201. Therefore, the heat exchange between the heater 200 and the reaction sample takes place between the side wall 2021 and the bottom 2022 of the groove 201, which increases the contact area between the heater 200 and the reaction sample and improves the heat transfer efficiency.

[0109] In this embodiment, the depth of the groove 201 is 0.5 mm, meaning that when the reaction sample fills the receiving cavity 21, the thickness of the reaction sample is 0.5 mm. A cover plate 29 can be connected to the top of the groove 201 to confine the reaction sample within the receiving cavity 21.

[0110] In the scheme where the reaction sample is in direct contact with the heater 200, there is no detachable interface between the heater 200 and the reaction sample, thus having a smaller thermal resistance. This allows for faster temperature control while achieving rapid heat conduction.

[0111] like Figure 3 As shown, in other alternative embodiments, the receiving cavity 21 includes a bottom wall 22, and a groove 201 is formed on the surface of the heater 200, with at least a portion of the wall surface 202 of the groove 201 in close contact with the bottom wall 22. Alternatively, the bottom wall 22 may be in close contact with at least a portion of the upper surface of the heater 200. Because the bottom wall 22 and the heater 200 are in close contact, no air layer is formed between them, thus allowing for rapid heat transfer. However, compared to the scheme where the reaction sample is in direct contact with the heater 200, this adds an extra heat transfer layer, the bottom wall 22, which correspondingly reduces the heat transfer efficiency.

[0112] To ensure close contact between the bottom wall 22 and the heater 200, the bottom wall 22 and the heater 200 can optionally be injection molded. Of course, other methods can also be used to achieve close contact, which will not be elaborated here. Specifically, the bottom wall 22 is in close contact with the heat spreader layer 241 of the heater 200.

[0113] Of course, in other alternative embodiments, a flexible thermal conductive element 203 can be provided between the bottom wall 22 and the heater 200. The flexible thermal conductive element 203 can ensure a more snug fit with the bottom wall 22 and the heater 200, so that both the bottom wall 22 and the heater 200 are in close contact with the flexible thermal conductive element 203, thereby avoiding the formation of an air layer. The flexible thermal conductive element 203 can be thermally conductive silicone or the like.

[0114] like Figure 1-5As shown, optionally, the heater 200 includes a heating element 23, and a power supply is connected to the heating element 23. The heating element 23 is a controllable heating source inside the heater 200 and can be a resistor, such as a thin resistance wire structure made of copper. The heating power is controlled by controlling the current flowing through the resistor, thereby achieving temperature control. In other optional embodiments, the heating element 23 can also adopt a coil structure or use electromagnetic induction heating through ferromagnetic materials, etc.

[0115] like Figure 4 As shown, the heater 200 may further include an upper conductive assembly 24 and a lower conductive assembly 27, with the heating element 23 sandwiched between the upper conductive assembly 24 and the lower conductive assembly 27. The upper conductive assembly 24 and the lower conductive assembly 27 have the functions of conducting heat and insulation.

[0116] like Figure 1 and Figures 4-5 As shown, the heater 200 includes a heat spreader layer 241. Specifically, the upper conductive assembly 24 may also include a heat spreader layer 241. The heat spreader layer 241 is in direct contact with the reaction sample in the receiving cavity 21 or in close contact with the bottom wall 22. The heat spreader layer 241 can ensure uniform heat conduction in both the longitudinal and transverse directions (i.e., the thickness direction of the reaction sample and the surface perpendicular to the thickness direction), ensuring the temperature uniformity of the sample liquid. Optionally, the heat spreader layer 241 is made of a conductive material or an insulating material, such as aluminum, copper, or other conductive materials, or high thermal conductivity ceramics or other insulating materials.

[0117] like Figure 1 As shown, when the heat spreader 241 is made of insulating material, the heat spreader 241 is adjacent to the heating element 23. At this time, the number of layers of the reaction assembly 2 can be reduced, the time for heat transfer from the heating element 23 to the reaction assembly 2 can be shortened, and the time required for heat dissipation of the reaction assembly 2 can be shortened.

[0118] like Figure 4 As shown, when the heat spreader 241 is made of a conductive material, the upper conductive assembly 24 further includes an insulating layer 242 located between the heating element 23 and the heat spreader 241. The insulating layer 242 is made of a material with high thermal conductivity and is used to provide electrical isolation between the heating element 23 and the conductive heat spreader 241. Optionally, the insulating layer 242 is made of a material with a thermal conductivity >1 W / mK; in this embodiment, the thermal conductivity of the insulating layer 242 is 3 W / mK.

[0119] like Figure 1 and Figures 4-5As shown, the lower conductive component 27 also includes an insulating thermal resistance layer 271. The insulating thermal resistance layer 271 possesses certain thermal resistance and insulation properties. Besides insulating the heating element 23, the insulating thermal resistance layer 271 also forms a longitudinal thermal resistance. The magnitude of the thermal resistance can be designed through material selection and thickness selection. Typically, the thermal resistance of this layer is much greater than that of other layers in the structure; therefore, the insulating thermal resistance layer 271 is the main source of thermal resistance for heat dissipation and cooling of the reaction component 2 to the cooling mechanism 1 (which will be described in detail below). The insulating thermal resistance layer 271 is one of the main factors affecting the thermal performance of the reaction component 2.

[0120] Optionally, the lower conductive assembly 27 further includes a heat-conducting layer 272, which is located on the side of the insulating thermal resistance layer 271 away from the heating element 23. Further, the heat-conducting layer 272 is the outermost layer of the lower conductive assembly 27, and it is in direct contact with the cooling mechanism 1. The heat-conducting layer 272 is made of metals such as copper or other materials with high thermal conductivity. Due to cost control or processing technology limitations, point contact between the surfaces of the lower conductive assembly 27 and the cooling mechanism 1 is unavoidable. When the outermost layer of the lower conductive assembly 27 is the heat-conducting layer 272, even if there is point contact between the heat-conducting layer 272 and the cooling mechanism 1, the heat-conducting layer 272 can still distribute heat evenly throughout the entire heat-conducting layer 272 due to its good conductivity, thereby ensuring a uniform heat distribution in the other layers of the lower conductive assembly 27.

[0121] Preferably, in this embodiment, the heating element 23 is a resistor, and there is a specific relationship between resistance and temperature. Therefore, while heating, the real-time resistance change of the heating element 23 is measured, and the average temperature of the heating element 23 is derived through the temperature coefficient of resistance and the nominal resistance value. This real-time temperature reading reflects the current temperature of the reaction component 2 without delay, which can be used for rapid feedback control of the reaction component 2 and the temperature of the reaction sample. Compared with the prior art, it can control the sample temperature more accurately and improve the overall reaction speed of the temperature control system.

[0122] The disadvantage of this method is that for resistors of the same type, such as copper wire resistors, the nominal resistance value and temperature coefficient of resistance may differ slightly between resistors. This can lead to a slight difference between the temperature coefficient of resistance and the nominal resistance value between individual heating elements 23, potentially causing temperature measurement errors. Therefore, preferably, as... Figure 1 and Figures 4-8 As shown, the heater 200 provided in this embodiment may also include a temperature calibration unit 25 for the temperature detection unit to detect the temperature, thereby enabling the reaction component 2 to be controlled by a dual temperature measurement method of resistance temperature measurement and temperature calibration of the reaction component 2.

[0123] Optionally, the temperature detection unit can be a contact temperature detection unit or a non-contact temperature detection unit 5.

[0124] like Figure 1 , Figures 4-6 , Figure 8 and Figure 10 As shown, when the temperature of the temperature calibration unit 25 is detected by contact or non-contact means, if the temperature detection unit is a contact temperature detection unit or a non-contact temperature detection unit 5, the temperature calibration unit 25 is connected to the upper conduction component 24 or the lower conduction component 27 so that the temperature of the heating element 23 is conducted to the temperature calibration unit 25.

[0125] like Figure 7 As shown, when the temperature detection unit is a non-contact temperature detection unit 5, the number of calibration units can be one or two, as long as the temperature of the temperature calibration unit 25 can be detected by the non-contact temperature detection unit 5.

[0126] like Figure 4 and Figure 6 As shown, when the temperature detection unit is a contact-type temperature detection unit, the two first contacts of the contact-type temperature detection unit are in contact with the two temperature calibration parts 25 respectively, and there is no electrical conductivity between the two temperature calibration parts 25. At this time, as... Figure 6 As shown, optionally, the nucleic acid amplification device may further include external electrical contact points 251 and electrical connection leads 252. The number of external electrical contact points 251 and electrical connection leads 252 may both be two. The two external electrical contact points 251 are located on opposite sides of the two temperature calibration units 25. One external contact point is electrically connected to one temperature calibration unit 25 through one electrical connection lead 252, and the other external contact point is electrically connected to the other temperature calibration unit 25 through another electrical connection lead 252.

[0127] Combination Figure 5 and Figure 10The heat from the upper conductive component 24 (such as the heat-spreading layer 241 of the upper conductive component 24 that contacts the patch 261 of the fast conductive part 26 (described in detail below)) 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 external electrical connection contact point 251 via the electrical connection lead 252. The diameter of the electrical connection lead 252 is smaller than that of the temperature calibration part 25 and the external electrical connection contact point 251, thereby reducing the heat loss generated by the temperature calibration part 25 through the electrical connection lead 252. Therefore, the temperature calibration part 25 can... The temperature of the upper conductive component 24 (such as the heat-spreading layer 241 of the upper conductive component 24 that contacts the patch 261) is well reflected. The temperature detection unit achieves good electrical and thermal contact with the temperature calibration unit 25 through solder joints. When the temperature of the upper conductive component 24 (such as the heat-spreading layer 241 of the upper conductive component 24 that contacts the patch 261) changes, the temperature detection unit can quickly and accurately sense the temperature change. The temperature change causes the resistance of the temperature detection unit to change. By detecting the resistance change of the temperature detection unit in real time at the external electrical connection contact point 251, real-time temperature detection can be achieved.

[0128] Specifically, such as Figure 4 and Figure 5 As shown, the temperature calibration unit 25 is located on the side of the lower conduction assembly 27 away from the heating element 23, to facilitate fixing the temperature calibration unit 25. Combined with Figure 6 and Figure 7 Furthermore, a receiving groove 275 is formed on the side of the lower conductive component 27 away from the heating element 23. The temperature calibration part 25 is located in the receiving groove 275 and connected to the bottom of the receiving groove 275. Optionally, the receiving groove 275 penetrates the heat-conducting layer 272, and the bottom of the receiving groove 275 is an insulating thermal resistance layer 271. The temperature calibration part 25 is connected to the insulating thermal resistance layer 271. In other optional embodiments, the receiving groove 275 may not penetrate the heat-conducting layer 272, and the temperature calibration part 25 may be connected to the heat-conducting layer 272. The receiving groove 275 can prevent the temperature calibration part 25 from protruding from the lower conductive component 27, thereby maintaining the flatness of the lower surface of the reaction component 2 and facilitating the stable placement of the reaction component 2. Of course, in yet another embodiment, the lower conductive component 27 may not have a receiving groove 275, and the temperature calibration part 25 may be connected to the lower surface of the lower conductive component 27.

[0129] In yet another alternative embodiment, such as Figure 8 As shown, the temperature calibration unit 25 is connected to the side of the upper conductive assembly 24 near the lower conductive assembly 27. For example, the temperature calibration unit 25 is connected to the lower surface of the insulating heat spreader 241 (e.g., Figure 8As shown, when the heat spreader 241 is insulated, the insulation layer 242 may not be provided. Therefore, the temperature calibration unit 25 may be directly connected to the lower surface of the heat spreader 241 or to the lower surface of the insulation layer 242, so that the temperature of the heating element 23 can be quickly transferred to the temperature calibration unit 25.

[0130] The lower conduction assembly 27 may also have a first through hole 273 facing the temperature calibration section 25, so that the temperature sensing element can detect the temperature of the temperature calibration section 25. The lower surface of the upper conduction assembly 24 and the upper surface of the lower conduction assembly 27 are closest to the heating element 23, and their temperatures are closest to the temperature of the heating element 23 first. Therefore, by connecting the temperature calibration section 25 to the side of the upper conduction assembly 24 near the lower conduction assembly 27, the temperature sensing unit can detect the temperature closest to the heating element 23 more quickly.

[0131] like Figure 1 and Figure 4 As shown, optionally, the temperature detection unit can be a first contact temperature detection unit 3, and the reaction assembly 2 can include a first contact temperature detection unit 3 connected to the temperature calibration unit 25 and used to measure the temperature at the temperature calibration unit 25. The first contact temperature detection unit 3 can be a temperature sensor or the like. The first contact temperature detection unit 3 can be connected to the temperature calibration unit 25 by means of welding or the like, and after the reaction assembly 2 is used, the first contact temperature detection unit 3 can be discarded along with the reaction assembly 2.

[0132] When the temperature of the temperature calibration unit 25 is detected using a non-contact method, such as when the temperature detection unit is a non-contact temperature detection unit 5 such as an infrared temperature measurement unit, the temperature calibration unit 25 can be configured in other ways besides the methods described above. Figure 9 As shown, the temperature calibration unit 25 can be part of the upper conduction component 24 or the lower conduction component 27. This eliminates the need for an additional connection to the temperature calibration unit 25; only a position for the temperature calibration unit 25 needs to be reserved in the upper conduction component 24 or the lower conduction component 27, allowing the non-contact temperature detection unit 5 to align with and detect the temperature at that position. Preferably, the lower conduction component 27 has a second through hole 274 along the thickness direction of the reaction component 2, and the surface of the upper conduction component 24 facing the second through hole 274 is the temperature calibration unit 25. The lower surface of the upper conduction component 24 and the upper surface of the lower conduction component 27 are closest to the heating element 23, and their temperatures are closest to the temperature of the heating element 23 first. Therefore, by detecting the temperature of the lower surface of the upper conduction component 24, the temperature detection unit can detect the temperature closest to the heating element 23 more quickly.

[0133] Of course, in other alternative embodiments, the temperature calibration unit 25 can be the lower surface of the thermally conductive layer 272 to simplify the structure of the reaction component 2.

[0134] like Figure 4 and Figure 5 As shown, to shorten the time it takes for the temperature of the temperature calibration unit 25 to match the temperature of the heating element 23, the reaction assembly 2 may optionally include a rapid conduction unit 26, which conducts heat from the heating element 23 to the temperature calibration unit 25. Specifically, in this embodiment, the heat from the heating element 23 is indirectly conducted to the temperature calibration unit 25. For example, when the heating element 23 heats the heat spreader 241, the heat from the heat spreader 241 is conducted to the temperature calibration unit 25 through the rapid conduction unit 26. Thus, the temperature calibration unit 25 accurately reflects the temperature of the heat spreader 241, and the temperature detection unit can accurately measure the temperature of the heat spreader 241. Furthermore, since the thickness of the reaction sample is very small, the temperature of the reaction sample is basically the same as the temperature of the heat spreader 241. Therefore, the temperature of the reaction sample can be obtained by detecting the temperature of the temperature calibration unit 25.

[0135] Preferably, one side of the rapid conduction section 26 is connected to the side of the upper conduction assembly 24 near the heating element 23 or to the side of the lower conduction assembly 27 near the heating element 23, and the other side is connected to the temperature calibration section 25. The lower surface of the upper conduction assembly 24 and the upper surface of the lower conduction assembly 27 are closest to the heating element 23, and their temperatures are closest to the temperature of the heating element 23 first. Therefore, the arrangement of the rapid conduction section 26 allows the temperature of the rapid conduction section 26 to reach the same temperature as the heating element 23 in the shortest possible time. Optionally, the rapid conduction section 26 is made of a material with high thermal conductivity, such as metals like copper or aluminum, or thermally conductive ceramics. The thermal conductivity of the rapid conduction section 26 is particularly superior to that of the lower conduction assembly 27, so as to quickly transfer heat to the temperature calibration section 25.

[0136] The rapid conduction section 26 includes a patch 261 and one or more second guide posts 262. The patch 261 is attached to the side of the upper conduction assembly 24 near the heating element 23 or to the side of the lower conduction assembly 27 near the heating element 23. One end of the one or more second guide posts 262 is connected to the patch 261, and the other end passes through the lower conduction assembly 27 and is connected to the temperature calibration section 25. The lower surface of the upper conduction assembly 24 and the upper surface of the lower conduction assembly 27 are closest to the heating element 23, and their temperatures are closest to the temperature of the heating element 23 first. Therefore, the arrangement of the patch 261 can make the temperature of the rapid conduction section 26 match the temperature of the heating element 23 as quickly as possible. The patch 261 can increase the contact area between the rapid conduction section 26 and the upper conduction assembly 24 or the lower conduction assembly 27, thereby improving the conduction efficiency. The cross-sectional area of ​​the second guide post 262 can be smaller than that of the patch 261, allowing for rapid heat conduction from the patch 261 to the temperature calibration unit 25. Simultaneously, the volume of the second guide post 262 can be minimized, thereby reducing the impact of the rapid heat conduction unit 26 on the lower conductive assembly 27 and ensuring that the insulating thermal resistance layer 271 generates the required thermal resistance as designed. Optionally, the patch 261 and the second guide post 262 can be made of a material with high thermal conductivity, such as copper. When the patch 261 and the second guide post 262 need to be insulating materials to prevent short circuits in the reaction assembly 2, the patch 261 or the second guide post 262 can be made of a material with high thermal conductivity, such as ceramic.

[0137] It is understandable that the temperature calibration unit 25 can be configured in a one-to-one correspondence with the patch 261, and two temperature calibration units 25 can also be connected to one patch 261. One temperature calibration unit 25 can be connected to one second guide post 262, and to improve the temperature uniformity of the temperature calibration unit 25, the temperature calibration unit 25 can also be connected to multiple second guide posts 262.

[0138] like Figure 10 As shown, in other optional embodiments, the rapid conduction section 26 may include one or more first guide posts 263. The rapid conduction section 26 does not have a patch 261. One end of the one or more first guide posts 263 is attached to the side of the upper conduction assembly 24 near the heating element 23, and the other end of the one or more first guide posts 263 is connected to the temperature calibration section 25. The first guide posts 263 do not affect other structures of the reaction assembly 2 and can quickly transfer heat to the temperature calibration section 25. Optionally, the first guide posts 263 are metal posts with high thermal conductivity, such as copper posts. One temperature calibration section 25 can be connected to one first guide post 263. To improve the temperature uniformity of the temperature calibration section 25, the temperature calibration section 25 can also be connected to multiple first guide posts 263.

[0139] Optionally, the lower conductive assembly 27 has 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 facilitates the placement of the first guide post 263 or the second guide post 262 without affecting the continuity of the lower conductive assembly 27, thus ensuring the performance of the lower conductive assembly 27. Optionally, the first guide post 263 or the second guide post 262 does not contact the inner wall of the third through hole to avoid heat conduction between the first guide post 263 or the second guide post 262 and the lower conductive assembly 27, thereby affecting the temperature of the temperature calibration section 25.

[0140] like Figure 7 As shown, to facilitate obtaining the resistance of the heating element 23 and supplying power to the heating element 23, optionally, a plurality of second contacts 20 are provided on the side of the reaction assembly 2 away from the receiving cavity 21. The second contacts 20 are electrically connected to the heating element 23. The current and voltage of the heating element 23 can be obtained through the second contacts 20, and thus the resistance value of the heating element 23 can be obtained.

[0141] In this embodiment, the second contact 20 enables the reaction component 2 to perform its own temperature measurement function. Compared with the traditional structure that can only measure temperature through an external temperature measuring unit, this embodiment can directly measure the temperature of the reaction component 2 itself, thus making the temperature measurement more accurate and faster, and improving the accuracy and control speed of the temperature control system.

[0142] In existing technologies, temperature detection units such as temperature sensors are used to detect the temperature of the reaction component 2. However, since it takes time for heat to transfer from the reaction component 2 to the temperature detection unit, the temperature measurement result under normal circumstances will have a temperature measurement delay of 1-2 seconds. During rapid heating and cooling, the temperature change of the reaction component 2 can reach more than 30°C in 1-2 seconds. Therefore, it is relatively difficult to control the reaction component 2 through the temperature detection unit during rapid heating and cooling. This embodiment does not rely entirely on the temperature value measured by the uncalibrated resistance thermometry method, nor does it rely entirely on the temperature detected by the temperature detection unit to control the reaction component 2. Instead, it combines the two by using the temperature calibration unit 25 of the reaction component 2 to measure the temperature and the heating element 23 to measure the temperature using the resistance thermometry method. This allows for rapid and accurate temperature control of the reaction component 2, achieving the goal of accurate temperature control. This overcomes the problems of temperature detection delay and large temperature measurement error caused by commonly used temperature detection methods in the prior art.

[0143] This embodiment also provides a nucleic acid amplification device, which includes the reaction component 2 as described above. The nucleic acid amplification device provided in this embodiment can improve amplification efficiency and shorten detection time.

[0144] like Figure 3As shown, the nucleic acid amplification device also includes a cooling mechanism 1, which is used to cool the reaction sample in the containment cavity 21 so that the reaction sample can complete the thermal cycle.

[0145] Optionally, the cooling mechanism 1 is located on the side of the reaction assembly 2 away from the receiving cavity 21. When cooling the reaction sample, the cooling mechanism 1 first cools the part of the reaction assembly 2 located between the reaction sample and the cooling mechanism 1.

[0146] Optionally, the cooling mechanism 1 cools the reaction assembly 2 via fluid flow, which can be gas or liquid. Specifically, as shown... Figure 3 As shown, the cooling mechanism 1 cools the reaction assembly 2 through fluid flow. Optionally, the cooling mechanism 1 includes a cooling body 12, within which a cooling channel 11 is provided to allow the cooling medium to flow. The cooling medium can be water or other liquids.

[0147] like Figure 11 As shown, the cooling mechanism 1 can also cool the reaction assembly 2 by jetting fluid. Optionally, the cooling medium can be water or gas, etc. Figure 11 (The middle arrow indicates a portion of the flow direction of the cooling medium). For example, cooling mechanism 1 may include a pump and a spray assembly. The spray assembly is connected to the pump, the pump pumps the high-pressure cooling medium into the spray assembly, and the spray assembly sprays the cooling medium toward reaction assembly 2.

[0148] In other alternative embodiments, the cooling mechanism 1 may not use a flowable or jet-like medium to cool the reaction component 2, but may instead use a solid cooling method to cool the reaction component 2, such as the cooling mechanism 1 including a semiconductor cooler.

[0149] like Figure 12 and Figure 13 As shown, optionally, the cooling mechanism 1 has a clearance portion 13 on the side near the reaction component 2. Specifically, the clearance portion 13 is used to avoid the temperature calibration unit 25. The clearance portion 13 can be a groove or hole provided on the cooling mechanism 1. The clearance portion 13 can prevent the cooling mechanism 1 from affecting the temperature of the temperature calibration unit 25, ensuring that the temperature calibration unit 25 accurately reflects the temperature of the heating element 23.

[0150] When the temperature detection unit is the first contact temperature detection unit 3, the avoidance part 13 can also avoid the first contact temperature detection unit 3.

[0151] like Figure 13 and Figure 14As shown, the temperature detection unit in this embodiment may not be the first contact temperature detection unit 3, but rather the second contact temperature detection unit 4. Specifically, the nucleic acid amplification device includes the second contact temperature detection unit 4, which can be separated from or in contact with the temperature calibration unit 25. When it is in contact with the temperature calibration unit 25, it can measure the temperature of the temperature calibration unit 25.

[0152] Specifically, the second contact temperature detection unit 4 can be positioned at the clearance portion 13 to fully utilize space and ensure easy contact between the temperature detection unit and the temperature calibration unit 25. The contact between the second contact temperature detection unit 4 and the temperature calibration unit 25 is elastic; for example, the second contact temperature detection unit 4 can be connected to the cooling mechanism 1 via a spring to achieve elastic contact. Since the second contact temperature detection unit 4 is not discarded with the reaction assembly 2, the cost of the reaction assembly 2 and the detection cost can be reduced.

[0153] The nucleic acid amplification device also includes a resistance detection unit 6, which is used to detect the temperature of the heating element 23. Specifically, the resistance detection unit 6 measures the resistance of the heating element 23 using a four-wire method. The resistance detection unit 6 can be electrically connected to the second contact 20 to detect the current and voltage of the heating element 23. Optionally, the resistance detection unit 6 is located on the side of the cooling mechanism 1. The position of the second contact 20 facilitates stable contact with the resistance detection unit 6, and allows the resistance detection unit 6 to be located on the side of the cooling mechanism 1, making the nucleic acid amplification device more compact.

[0154] Optionally, the cooling mechanism 1 remains in continuous contact with the reaction assembly 2, generating heat through the heating element 23 and the heat spreader 241 to heat the reaction sample. Simultaneously, heat is transferred downwards through the insulating thermal resistance layer 271 and dissipated through the interface between the thermally conductive layer 272 and the cooling mechanism 1. When the heat dissipation power equals the heat generation power, the system achieves thermal equilibrium, and the reaction sample can be maintained at a specific temperature. By changing the heating power of the internal heating element 23, the temperature of the reaction sample at thermal equilibrium can be adjusted, thereby achieving dynamic adjustment of the reaction sample temperature.

[0155] In this embodiment, the reaction component 2 adopts a thin-layer structure with integrated heating to achieve rapid temperature change and thermal equilibrium of the reaction sample.

[0156] The heating rate of the reaction sample is proportional to the product of its heat capacity and heating thermal resistance. The heat capacity of the reaction sample includes the heat capacity of the liquid and the heat capacity of the auxiliary structures of the heating element 23 near the reaction sample, such as the conductive component 24 and the cover plate 29. The heating thermal resistance includes the thermal resistance of the interfaces and materials from the heating element 23 to the reaction sample, as well as the equivalent thermal resistance of heat transfer within the reaction sample. Under the same heating amplitude (the same temperature change), the smaller the product of the heat capacity and heating thermal resistance of the reaction sample, the faster the heating rate. In this embodiment, since the heating element 23 is a thin-layer structure (without a traditional heating metal block) bonded to the heat spreader 241, and the heat spreader 241 is tightly bonded to the reaction sample and the cover plate 29, the heat capacity of the reaction sample is much smaller than that of a traditional structure. Simultaneously, since the reaction assembly is an integral structure with no contact thermal resistance, and the structure of the heating element 23 near the reaction sample uses a high thermal conductivity material such as metal, the reaction sample layer is thin (the longitudinal thermal resistance of the reaction sample is small), resulting in a very small overall heating thermal resistance. Therefore, the reaction assembly provided in this embodiment can achieve a heating rate much higher than that of conventional PCR heating schemes.

[0157] The cooling rate of the reaction sample is proportional to the product of its heat capacity and thermal resistance. The heat capacity includes the heat capacity of the liquid and the heat capacity of auxiliary structures near the reaction sample on the heating element 23, such as the conductive component 24 and the cover plate 29. The thermal resistance includes the thermal resistance from the heating element 22 to the interface and materials of the cooling mechanism, as well as the equivalent thermal resistance of heat transfer within the reaction sample. Under the same cooling rate (the same temperature change during cooling), the smaller the product of the heat capacity and thermal resistance, the faster the cooling rate. In this embodiment, because the heating element 23 is a thin-layer structure (without a traditional heating metal block) bonded to the heat spreader 241, and the heat spreader 241 is tightly bonded to the reaction sample and the cover plate 29, the heat capacity of the reaction sample is much smaller than that of the traditional structure. Simultaneously, since the only contact surface is between the reaction component 2 and the cooling mechanism 1, the thermal resistance of the insulating thermal resistance layer 271 can be designed to be relatively small. The reaction component is an integral structure, the heat spreader 241 uses a high thermal conductivity material such as metal, and the reaction sample is relatively thin (low longitudinal thermal resistance of liquids), resulting in a very low overall heat dissipation thermal resistance. Therefore, the nucleic acid amplification device of this embodiment can achieve a cooling rate much higher than that of conventional PCR heating schemes. Specifically, the nucleic acid amplification device of this embodiment can achieve heating and cooling rates of ~20℃ / second, or even higher.

[0158] Selection of Insulating Thermal Resistance Layer 271: In the design of reaction components 2 of the same size, the temperature control performance of the sample can be optimized through the design of the insulating thermal resistance layer 271. When the thermal resistance of the insulating thermal resistance layer 271 is large, less heat dissipation occurs, and the heating power of the heating element 23 is lower when achieving the same thermal equilibrium temperature, which is beneficial to reducing the total power consumption of the nucleic acid amplification device. However, because the thermal resistance of the insulating thermal resistance layer 271 is large, the heat dissipation thermal resistance is increased, which will reduce the cooling rate of the reaction sample. Similarly, when the thermal resistance of the insulating thermal resistance layer 271 is small, heat dissipation is faster, and the heating power of the heating element 23 is higher when achieving the same thermal equilibrium temperature, resulting in a higher total power consumption of the nucleic acid amplification device. However, at the same time, because the thermal resistance of the insulating thermal resistance layer 271 is reduced, the cooling rate of the reaction sample will be increased, which is beneficial to shortening the overall time of the PCR process. The insulating thermal resistance layer 271 can be adjusted in terms of both material and thickness to meet different design requirements. In typical designs, a thin layer of 0.1-0.3 mm thickness can be used, with a material thermal conductivity of 0.2-0.5 W / mK. For a reaction sample with a thickness of 0.5 mm and corresponding auxiliary structure design, a cooling rate of ~20℃ / second can be achieved, with an average PCR temperature control power of about 30-50 W.

[0159] Example 2

[0160] like Figure 15 As shown, the nucleic acid amplification device in this second embodiment is basically the same as that in the first embodiment, except that the temperature detection unit in this second embodiment is not the first contact temperature detection unit 3, and the temperature detection unit is not the non-contact temperature detection unit 5. In this second embodiment, the nucleic acid amplification device includes a non-contact temperature detection unit 5, which is used to measure the temperature at the temperature calibration unit 25.

[0161] Specifically, the non-contact temperature detection unit 5 can be located at the clearance portion 13 to make full use of space and ensure that the non-contact temperature detection unit 5 can detect the temperature of the temperature calibration portion 25. Since the non-contact temperature detection unit 5 is not discarded with the reaction assembly 2, the cost of the reaction assembly 2 and the detection cost can be reduced.

[0162] Example 3

[0163] This third embodiment provides a nucleic acid amplification control method, which can be executed using the reaction component 2 or nucleic acid amplification device from embodiments one and two.

[0164] In the prior art, temperature detection units such as temperature sensors are used to detect the temperature of the reaction component. However, since it takes a certain amount of time for heat to be transferred from the reaction component 2 to the temperature detection unit, the detection result measured by the temperature detection unit will have a temperature measurement delay of 1 to 2 seconds under normal circumstances. During the rapid heating and cooling process, the temperature change of the reaction component 2 can reach more than 30°C in 1 to 2 seconds. Therefore, it is relatively difficult to control the reaction component 2 through the temperature detection unit during the rapid heating and cooling process.

[0165] There is a specific relationship between the resistance of heating element 23 and its temperature. Therefore, while heating, the real-time resistance change of heating element 23 in reaction assembly 2 is measured, and the average temperature of heating element 23 is derived by comparing the resistance temperature coefficient with the resistance value at the nominal temperature (the nominal resistance value refers to the actual resistance value claimed (or marked) at this temperature, which can be arbitrarily selected according to requirements). This temperature reading reflects the current temperature of reaction assembly 2 in real time without delay, thus enabling rapid feedback control of the temperature of reaction assembly 2 and the reaction sample. The disadvantage of this method is that for resistors of the same type, such as copper wire resistors, there are slight differences in the nominal resistance value and resistance temperature coefficient between resistors, resulting in slight differences in the resistance temperature coefficient and nominal resistance value between individual heating elements 23, which may cause temperature measurement errors.

[0166] In this embodiment, to avoid the shortcomings of the above two temperature measurement methods and improve the accuracy of temperature control, such as... Figure 16 and Figure 17 As shown, the nucleic acid amplification control method includes controlling the reaction component 2 through a dual temperature measurement method, namely resistance temperature measurement and temperature calibration of the reaction component 2.

[0167] This embodiment does not rely entirely on the temperature value measured by the uncalibrated resistance thermometry method, nor does it rely entirely on the temperature control reaction component 2 detected by the temperature detection unit. Instead, it overcomes the above-mentioned defects by combining the two and using a method of calibrating the temperature of the reaction component 2, thereby enabling rapid and accurate control of the temperature of the reaction component 2 and achieving the purpose of accurate temperature control.

[0168] Controlling reaction component 2 using a dual temperature measurement method, including resistance temperature measurement and temperature calibration of reaction component 2, includes:

[0169] The temperature calibration value of the reaction component 2 is measured by the temperature detection unit, and the temperature value corresponding to the resistance value of the reaction component 2 is obtained through the temperature calibration value. That is, the temperature detection unit detects the temperature calibration value of the reaction component 2, and by combining the temperature calibration value with the voltage and current of the heating element 23 of the reaction component 2 detected by the resistance thermometry method, the temperature measurement by the resistance thermometry method is calibrated. Preferably, the temperature detection unit detects the temperature at the temperature calibration section 25 of the reaction component 2 to obtain the temperature calibration value. It can be understood that the temperature calibration value is a temperature value selected manually, and this temperature calibration value is the current actual temperature of the reaction component 2.

[0170] Specifically, by using the temperature calibration value and combining it with the resistance value of the heating element 23 at that temperature calibration value, the resistance temperature coefficient and nominal resistance value of the specific heating element 23 can be obtained. When using the resistance temperature measurement method to measure the temperature later, the actual temperature of the reaction component 2 can be obtained through the resistance temperature coefficient and the nominal resistance value.

[0171] Temperature measurement is achieved through a dual method: resistance temperature measurement and calibration of the temperature of reaction component 2. The steps include the following:

[0172] At least two different temperature calibration values ​​are obtained; the first voltage and first current of the reaction component 2 at one temperature calibration value are detected, and the first resistance value of the reaction component 2 is obtained based on the first voltage and first current; the second voltage and second current of the reaction component 2 at another temperature calibration value are detected, and the second resistance value of the reaction component 2 is obtained based on the second voltage and second current.

[0173] Based at least the first resistance value, the second resistance value, and the temperature calibration value corresponding to the first resistance value and the temperature calibration value corresponding to the second resistance value, the resistance temperature coefficient and nominal resistance value of the reaction component 2 can be obtained.

[0174] The current and voltage of reaction component 2 are continuously monitored, and the reaction component 2 is controlled according to the temperature coefficient of resistance and the nominal resistance value.

[0175] Understandably, the at least two different temperature calibration values ​​obtained are read when the temperature detection unit and the temperature of reaction component 2 are essentially the same, and this temperature is considered the actual temperature of reaction component 2. At this time, the corresponding current and voltage of reaction component 2 are read, and the temperature coefficient of resistance and the nominal resistance value are deduced, thus achieving the calibration of the temperature coefficient of resistance and the nominal resistance value. Therefore, when the temperature of reaction component 2 is subsequently obtained based on the current and voltage of reaction component 2, it can be considered the accurate temperature of reaction component 2.

[0176] Furthermore, according to the formula R=R0(1+α) T T = T - T0, R is the resistance value of heating element 23 at temperature T, T0 is the nominal temperature, R0 is the nominal resistance value, and α is the temperature coefficient of resistance of the material. This calibrates the temperature coefficient of resistance and nominal resistance value of the reaction assembly 2. That is, the temperature detection unit measures the first temperature calibration value T1, and the resistance detection unit 6 detects the first voltage U1 and the first current I1 of heating element 23 at temperature T1. According to R = U / I, the resistance R1 of heating element 23 at temperature T1 can be obtained. Subsequently, the temperature detection unit measures the second temperature calibration value T2, and the resistance detection unit 6 detects the second voltage U2 and the second current I2 of heating element 23 at temperature T2. According to R = U / I, the resistance R2 of heating element 23 at temperature T2 can be obtained. Finally, according to two sets of two linear equations: R1 = R0(1 + α... T1) and R2=R0(1+α) T2 T 1= T1-T 0； T 2= T2-T 0； The specific values ​​of α and R0 are obtained. Then, by continuously measuring the voltage and current of reaction component 2, the values ​​are calculated according to the formula R = R0(1 + α). The temperature curve of reaction component 2 is obtained by T). Since the R0 and α values ​​in the formula are specific to the heating element 23, the temperature value can be obtained accurately.

[0177] It is understandable that when calibrating temperature values, the greater the difference between different temperature calibration values, the more accurate the obtained temperature coefficient of resistance and nominal resistance value will be. Therefore, optionally, the difference between two adjacent temperature calibration values ​​should not be less than 20°C.

[0178] It is understandable that temperature calibration values ​​can be detected throughout the entire nucleic acid amplification process, but only a few temperature calibration values ​​can be selected for temperature calibration. Optionally, the temperature value can be calibrated using at least two unequal temperature calibration values. For example, two, three, four, or more temperature calibration values ​​can be measured to calibrate the temperature value. Two temperature calibration values ​​can yield a temperature coefficient of resistance and a nominal resistance value; similarly, three or more temperature calibration values ​​can yield more than one temperature coefficient of resistance and more than one nominal resistance value. Using more than one temperature coefficient of resistance and more than one nominal resistance value can yield values ​​closer to the actual temperature coefficient of resistance and nominal resistance value, thereby further improving detection accuracy.

[0179] To overcome the problem of inaccurate temperature measurement caused by the 1-2 second temperature measurement delay of the temperature detection unit, and to ensure that the temperature calibration value reflects the current actual temperature of the reaction component 2, in this embodiment, at least two unequal temperature calibration values ​​are read after the temperature of the temperature detection unit matches the temperature of the reaction component 2. For example, after pausing for 1-2 seconds at a certain temperature, the temperature calibration value is read to ensure the accuracy of temperature calibration. Alternatively, if the temperature calibration value is obtained during the heating or cooling process, the heating or cooling rate is slowed down to ensure that the temperature of the temperature detection unit matches the temperature of the reaction component 2, or the temperature change is paused during the heating or cooling process, and the heating or cooling process is resumed after obtaining the temperature calibration value. If the temperature calibration value is obtained during the heat preservation stage, the temperature detected by the temperature detection unit is read after a certain heat preservation time, such as after 2 seconds, 3 seconds, or 8 seconds.

[0180] It is understandable that the first heating process in the nucleic acid amplification process is the process of heating to the pretreatment stage. The heating process, high-temperature denaturation stage, cooling process, low-temperature annealing stage, heating process, and suitable temperature extension stage are considered as one amplification cycle.

[0181] Optionally, the temperature calibration value used for calibrating the temperature value can be obtained before the nucleic acid amplification process, during the first temperature rise in the nucleic acid amplification process, and / or during the first amplification cycle in the nucleic acid amplification process. That is, it is preferable to use the calibration temperature value measured within the above time period to calibrate the temperature calibration value. Obtaining the temperature calibration value used for calibrating the temperature value during or before the first amplification cycle, and completing the temperature calibration during the first amplification cycle, can ensure the accuracy of subsequent temperature control.

[0182] Understandably, before the nucleic acid amplification process, the temperature detection unit detects the room temperature of reaction component 2, and this room temperature is used as one of the calibration temperature values. Alternatively, before the nucleic acid amplification process, the temperature of reaction component 2 can be changed to obtain the temperature calibration value required for calibration through the temperature detection unit. However, this increases the detection time because it takes time for the temperature of the temperature detection unit to match the temperature of reaction component 2. Therefore, it is preferable to combine the room temperature of reaction component 2 detected by the temperature detection unit with the temperature calibration value measured during the first temperature rise in the nucleic acid amplification process and / or during the first amplification cycle of the nucleic acid amplification process.

[0183] like Figure 16 and Figure 17As shown, optionally, before calibrating the temperature value, the temperature of the reaction component 2 is controlled according to a preset RT temperature curve. This avoids excessive temperature deviation in the reaction component 2 due to rapid temperature changes. After calibrating the temperature value, the temperature value detected by resistance thermometry continues to control the reaction component 2. Of course, in other optional embodiments, the temperature of the reaction component 2 can also be controlled by the temperature measured by the temperature detection unit before calibrating the temperature value. In this case, the temperature rise and fall rate is reduced during the heating and cooling stages to avoid excessively high or low temperatures.

[0184] To more clearly illustrate the nucleic acid amplification control method in this embodiment, combined with Figure 17 The diagram illustrates the calibration process of resistance thermometry using a temperature detection unit in a practical test. Before calibrating the temperature value, an initial RT temperature curve, i.e., a preset temperature curve, is established. Then, a very small current (e.g., less than 1 mA) is applied to the heating element 23 of the reaction component 2. The purpose of applying a very small current is to read the resistance of the heating element 23 without causing it to overheat.

[0185] First calibration: The temperature detection unit measures the first temperature calibration value T1, and the resistance detection unit detects the first voltage U1 and the first current I1 of the heating element 23 at temperature T1. According to R=U / I, the resistance R1 of the heating element 23 at temperature T1 can be obtained.

[0186] Second calibration: Subsequently, the temperature detection unit measures the second temperature calibration value T2, and the resistance detection unit 5 detects the second voltage U2 and the second current I2 of the heating element 23 at temperature T2. According to R=U / I, the resistance R2 of the heating element 23 at temperature T2 can be obtained.

[0187] Finally, based on the two sets of linear equations in two variables: R1=R0(1+α) T1) and R2=R0(1+α) T2), to obtain the specific values ​​of R0 and α, that is, to obtain the accurate RT curve, and then the temperature of the heating element 23 measured by the resistance temperature measurement method can be used as feedback for accurate temperature control.

[0188] Temperature calibration values ​​can be detected throughout the entire nucleic acid amplification process, allowing for multiple temperature calibrations in subsequent processes to further improve detection accuracy.

[0189] Continue to refer to Figure 17 When obtaining the first temperature calibration value, this temperature calibration value can be considered as the nominal resistance value. Therefore, based on this temperature calibration value, the formula R=R0(1+α) can be applied. The R0 in T) is corrected once to achieve the first calibration of the temperature value measured by the resistance thermometry method (e.g., Figure 17 The temperature curve measured by the resistance thermometry fluctuates after the first calibration. Subsequently, the temperature value obtained by the resistance thermometry or the temperature value measured by the temperature detection unit controls the temperature of reaction component 2 to the pretreatment stage. After the reaction component 2 is heated to the pretreatment stage and held for 8 seconds (not limited to 8 seconds, it can be any duration longer than 2 seconds but shorter than the pretreatment stage holding time), a second temperature calibration value is obtained for calibrating the temperature value. Based on the two temperature calibration values, the temperature value detected by the resistance thermometry is calibrated a second time, and the reaction component 2 is controlled to adjust to the accurate temperature. Then, the reaction component 2 continues to be controlled based on the temperature value obtained by the resistance thermometry. It can be understood that when the first temperature calibration value is room temperature, the temperature value measured by the resistance thermometry can be calibrated for the first time. Subsequently, a second temperature calibration value can be obtained at any temperature higher than room temperature, and the temperature value detected by the resistance thermometry can be calibrated a second time based on the two temperature calibration values, thus completing the calibration of the temperature value.

[0190] Because heat transfer from reaction component 2 to the temperature detection unit takes time, the temperature measurement result under normal circumstances will have a 1-2 second delay. During rapid heating and cooling, the temperature change of reaction component 2 can reach over 30°C within 1-2 seconds. Therefore, controlling reaction component 2 through the temperature detection unit during rapid heating and cooling is relatively difficult. Resistance thermometry can measure the real-time resistance change while heating and derive the average temperature of the resistor using the temperature coefficient of resistance and the nominal resistance value. This real-time, delay-free temperature reading reflects the current temperature of reaction component 2, thus enabling rapid temperature control of the sample. However, a drawback of resistance thermometry is that the temperature coefficient of resistance of a single resistor may slightly differ from the nominal resistance value, potentially causing temperature measurement errors. This embodiment overcomes these shortcomings by combining the two methods and calibrating the temperature of reaction component 2, thereby achieving rapid and accurate temperature control.

[0191] Although the present invention has been described in detail above with general descriptions, specific embodiments, and experiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. A reaction assembly, characterized by, The reaction assembly comprises a receiving cavity (21) for receiving a reaction sample and a heater (200) for heating the reaction sample; The heater (200) further comprises a heating element (23) and a temperature calibration portion (25) for detecting temperature of the temperature detecting unit, and a plurality of second contacts (20) are arranged on a side of the reaction assembly away from the receiving cavity (21) and electrically connected with the heating element (23), so that current and voltage of the heating element (23) can be obtained through the second contacts (20); The heater (200) further comprises an upper conducting assembly (24) and a lower conducting assembly (27), and the heating element (23) is clamped between the upper conducting assembly (24) and the lower conducting assembly (27), the upper conducting assembly (24) comprises a uniform heating layer (241), and the temperature calibration portion (25) is connected to the upper conducting assembly (24) or the lower conducting assembly (27); The reaction assembly further comprises a rapid conducting portion (26) for conducting heat of the heating element (23) to the temperature calibration portion (25).

2. The reaction assembly of claim 1, wherein, The heater (200) is in direct contact with the reaction sample in the receiving cavity (21).

3. The reaction assembly of claim 2, wherein, At least part of the upper surface of the heater (200) is in direct contact with the reaction sample in the receiving cavity (21), or A groove (201) is formed on the surface of the heater (200), and at least part of the wall surface (202) of the groove (201) is in direct contact with the reaction sample in the receiving cavity (21).

4. The reaction assembly of claim 1, wherein, The uniform heating layer (241) is in direct contact with the reaction sample in the receiving cavity (21).

5. The reaction assembly of claim 1, wherein, The temperature calibration portion (25) is connected to a side of the upper conducting assembly (24) close to the lower conducting assembly (27), and the lower conducting assembly (27) is provided with a first through hole (273) arranged opposite to the temperature calibration portion (25).

6. The reaction assembly of claim 1, wherein, The temperature calibration portion (25) is part of the upper conducting assembly (24) or the lower conducting assembly (27).

7. The reaction assembly of claim 6, wherein, The lower conducting assembly (27) is provided with a second through hole (274) along the thickness direction of the reaction assembly, and the surface of the upper conducting assembly (24) opposite to the second through hole (274) is the temperature calibration portion (25).

8. The reaction assembly of claim 1, wherein, One side of the rapid conducting portion (26) is connected to a side of the upper conducting assembly (24) close to the heating element (23) or a side of the lower conducting assembly (27) close to the heating element (23), and the other side is connected to the temperature calibration portion (25).

9. The reaction assembly of claim 1, wherein, The temperature calibration portion (25) is located on a side of the lower conducting assembly (27) away from the heating element (23).

10. The reaction assembly of claim 1, wherein, The lower conducting assembly (27) is provided with a receiving groove (275) on a side away from the heating element (23), and the temperature calibration portion (25) is located in the receiving groove (275) and connected to the groove bottom of the receiving groove (275).

11. The reaction assembly of claim 9, wherein, The rapid conducting part (26) comprises one or more first conducting columns (263), one end of which is attached to the upper conducting assembly (24) near the side of the heating element (23) or to the lower conducting assembly (27) near the side of the heating element (23), and the other end of which is connected to the temperature calibration part (25).

12. The reaction assembly of claim 9, wherein, The rapid conducting part (26) comprises a patch (261) and one or more second conducting columns (262), the patch (261) is attached to the upper conducting assembly (24) near the side of the heating element (23) or to the lower conducting assembly (27) near the side of the heating element (23), one end of the second conducting column (262) is connected to the patch (261), and the other end is arranged in the lower conducting assembly (27) and connected to the temperature calibration part (25).

13. The reaction assembly of claim 11 or 12, wherein, The lower conducting assembly (27) is provided with a third through hole, and the conducting column is arranged in the third through hole.

14. The reaction assembly of claim 1, wherein, The lower conducting assembly (27) further comprises an insulating thermal resistance layer (271).

15. The reaction assembly of claim 14, wherein, The lower conducting assembly (27) further comprises a thermal conductive layer (272), which is located on the side of the insulating thermal resistance layer (271) away from the heating element (23).

16. The reaction assembly of claim 1, wherein, The uniform heating layer (241) is made of conductive material or insulating material.

17. The reaction assembly of claim 16, wherein, When the uniform heating layer (241) is made of conductive material, the upper conducting assembly (24) further comprises an insulating layer (242) located between the heating element (23) and the uniform heating layer (241). When the uniform heating layer (241) is made of insulating material, the uniform heating layer (241) is adjacent to the heating element (23).

18. The reaction assembly of claim 1, wherein, The accommodating cavity (21) comprises a bottom wall (22). The bottom wall (22) is in close contact with at least part of the upper surface of the heater (200), or The surface of the heater (200) is formed with a groove (201), and at least part of the wall surface (202) of the groove (201) is in close contact with the bottom wall (22).

19. The reaction assembly of claim 18, wherein, A flexible thermal conductive element (203) is arranged between the bottom wall (22) and the heater (200).

20. The reaction assembly of claim 1, wherein, The accommodating cavity (21) has a flat structure.

21. The reaction assembly of claim 1, wherein, The cross section of the accommodating cavity (21) is polygonal, circular or elliptical.

22. The reaction assembly of any of claims 5, 1, 8-12, 14, 15, wherein, The reaction assembly comprises a first contact type temperature detection unit (3) connected to the temperature calibration part (25) and used for measuring the temperature at the temperature calibration part (25).

23. A nucleic acid amplification device, comprising: The reaction assembly (2) according to any one of claims 1-22.

24. A nucleic acid amplification device, characterized in that, The reaction assembly (2) according to any one of claims 5, 1, 8-15 and a second contact type temperature detection unit (4) capable of being separated from or in contact with the temperature calibration part (25) and capable of measuring the temperature of the temperature calibration part (25) when in contact with the temperature calibration part (25).

25. A nucleic acid amplification device, comprising: The nucleic acid amplification device comprises a non-contact temperature detection unit (5) and the reaction assembly (2) according to any one of claims 1, 5-19, wherein the non-contact temperature detection unit is used for measuring the temperature at the temperature calibration part (25).

26. The nucleic acid amplification device of any one of claims 23-25, wherein, The nucleic acid amplification device further comprises a cooling mechanism (1) used for cooling the reaction sample in the accommodating cavity (21).

27. The nucleic acid amplification device of claim 26, wherein, The cooling mechanism (1) is provided with a relief part (13) on one side close to the reaction assembly (2).

28. The nucleic acid amplification device of claim 26, wherein, The cooling mechanism (1) cools the reaction assembly (2) by fluid.

29. The nucleic acid amplification device of claim 28, wherein, The cooling mechanism (1) cools the reaction assembly (2) by spraying fluid or fluid flow.

30. The nucleic acid amplification device of claim 29, wherein, When the cooling mechanism (1) cools the reaction assembly (2) by fluid flow, the cooling mechanism (1) comprises a cooling body (12) provided with a cooling flow channel (11) for flowing cooling medium.

31. The nucleic acid amplification device of claim 26, wherein, The cooling mechanism (1) cools the reaction assembly (2) by solid cooling.

32. The nucleic acid amplification device of any one of claims 23-25, wherein, The nucleic acid amplification device further comprises a resistance detection unit (6) used for detecting the temperature of the reaction assembly (2).

33. The nucleic acid amplification device of claim 32, wherein, The resistance detection unit (6) obtains the temperature value of the reaction assembly (2) by detecting the resistance of the reaction assembly (2).

34. A method of nucleic acid amplification control, comprising: The nucleic acid amplification control method is performed by using the reaction assembly (2) according to any one of claims 1-22 or the nucleic acid amplification device according to claims 22-33. The nucleic acid amplification control method comprises controlling the reaction assembly (2) by resistance temperature measurement and temperature calibration of the reaction assembly (2).

35. The method of nucleic acid amplification control of claim 34, wherein, Controlling the reaction assembly (2) by resistance temperature measurement and temperature calibration of the reaction assembly (2) comprises: Measuring the temperature calibration value of the reaction assembly (2) by a temperature detection unit, and calibrating the temperature value obtained by the resistance value of the reaction assembly (2) by the temperature calibration value.

36. The method of nucleic acid amplification control of claim 35, wherein, Obtaining the resistance temperature coefficient and the nominal resistance value of the reaction assembly (2) according to the temperature calibration value, so as to calibrate the temperature value.

37. The method of nucleic acid amplification control of claim 35, wherein, Calibrating the temperature value by measuring the temperature calibration value before the nucleic acid amplification process, in the first heating process of the nucleic acid amplification process and / or in the first amplification cycle of the nucleic acid amplification process.

38. The nucleic acid amplification control method of claim 35, wherein, Calibrating the temperature value by using at least two different temperature calibration values.

39. The method of nucleic acid amplification control according to any one of claims 35-38, wherein, Controlling the reaction assembly (2) by resistance temperature measurement and temperature calibration of the reaction assembly (2) comprises the following steps: Obtaining at least two different temperature calibration values; detecting the first voltage and the first current of the reaction assembly (2) at one temperature calibration value, and obtaining the first resistance value of the reaction assembly (2) according to the first voltage and the first current; detecting the second voltage and the second current of the reaction assembly (2) at another temperature calibration value, and obtaining the second resistance value of the reaction assembly (2) according to the second voltage and the second current; According to the first resistance value, the second resistance value and the corresponding temperature calibration value, the resistance temperature coefficient and the resistance at the nominal temperature of the reaction component (2) are obtained; The current and voltage of the reaction component (2) are continuously detected, and the reaction component (2) is controlled according to the resistance temperature coefficient and the resistance at the nominal temperature.

40. The method of nucleic acid amplification control according to any one of claims 35-38, wherein, Before the temperature value is calibrated, the temperature of the reaction component (2) detected by the temperature detection unit controls the reaction component (2), or the temperature of the reaction component (2) is controlled according to the preset RT temperature curve of the reaction component (2).

41. The method of nucleic acid amplification control according to any one of claims 35-38, wherein, According to the formula: R = R0(1 + a T) the temperature curve of the reaction component (2) is determined from the formula, by continuously measuring the voltage and current of the reaction component (2), wherein R0 is the nominal resistance value and a is the temperature coefficient of resistance of the material.

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