Nucleic acid amplification systems, amplification methods, and detection methods; nucleic acid detection systems

Abstract

This disclosure provides a nucleic acid amplification system and its amplification and detection methods, as well as a nucleic acid detection system. The nucleic acid amplification system includes: a microfluidic cartridge (9) comprising an amplification section having an amplification cavity (90) for containing amplification reagents; at least two temperature control zones arranged along a first direction (x), the temperature control zones being configured to contain the amplification section, and at least a portion of the temperature control zones having adjustable temperatures; a temperature control component (2) configured to independently adjust the temperature of each temperature control zone to make the temperatures of each temperature control zone different; and a first driving component, drivenly connected to the amplification section and / or the temperature control component (2), configured to cause the amplification section and the temperature control component (2) to move relative to each other along the first direction (x), so that the amplification cavity (90) cyclically switches between at least two temperature control zones.

Description

Technical Field

[0001] This disclosure relates to the field of microfluidic detection technology, and in particular to a nucleic acid amplification system and its amplification and detection methods, as well as a nucleic acid detection system. Background Technology

[0002] Polymerase chain reaction (PCR) is a nucleic acid amplification method that mimics the natural DNA replication process in vitro. It can amplify a large amount of specific DNA fragments in a short time, thereby achieving highly sensitive detection. By adding samples (containing DNA template), nucleotides, primers, and DNA polymerase to the reactor, and controlling the temperature cycle, exponential amplification of DNA can be achieved. The PCR reaction mainly includes pre-denaturation, denaturation, annealing, and extension phases, with the denaturation-annealing-extension phase being repeated cyclically. Typically, one nucleic acid amplification consists of 30-50 PCR cycles. After each PCR cycle, the amount of nucleic acid molecules doubles. Throughout the PCR amplification process, the amount of DNA template increases exponentially with the number of cycles. Each stage of the PCR reaction requires appropriate temperature conditions and holding times to ensure the complete execution of the corresponding biological reaction.

[0003] Quantitative real-time PCR (qPCR) is currently the most commonly used clinical detection method. It involves adding fluorescent probes to the reaction system to indicate the reaction progress. The accumulation of amplified products is monitored by the accumulation of fluorescence signals, and the results are determined by fluorescence curves. Quantification can be achieved using Ct values ​​and standard curves. As the PCR cycle progresses, more and more double-stranded DNA is generated, leading to a continuous increase in fluorescence signals. Fluorescence signals can be detected in each cycle of the PCR amplification reaction, thus achieving real-time monitoring of the entire PCR amplification process.

[0004] Traditional nucleic acid amplification systems use a single temperature zone to control temperature cycling; however, the temperature change rate of a single temperature zone is low. To address this issue, nucleic acid amplification systems use high-power semiconductor cooling devices for temperature regulation to increase the temperature change rate. However, the instantaneous power of high-power semiconductor cooling devices increases significantly during regulation, generating a large amount of Joule heat, which affects the temperature stability of the nucleic acid amplification system. Summary of the Invention

[0005] The purpose of this disclosure is to provide a nucleic acid amplification system, amplification method, and detection method thereof, as well as a nucleic acid detection system, with the aim of improving the temperature change rate and temperature stability of the nucleic acid amplification system.

[0006] The first aspect of this disclosure provides a nucleic acid amplification system, comprising: A microfluidic cartridge includes an amplification section having an amplification cavity for containing amplification reagents; At least two temperature control zones are arranged along a first direction, and the temperature control zones are configured to accommodate the amplification section, with the temperature of at least a portion of the temperature control zones being adjustable. The temperature control components are configured to independently adjust the temperature of each temperature control zone, so that the temperature of each temperature control zone is different; and A first driving component, drivenly connected to the amplification section and / or the temperature control component, is configured to move the amplification section and the temperature control component relative to each other along a first direction, so that the amplification chamber cycles between at least two temperature control zones.

[0007] In some embodiments, at least two temperature control zones include a first temperature control zone and a second temperature control zone. The temperature of the first temperature control zone is in a first temperature range, and the temperature of the second temperature control zone is in a second temperature range. The temperature value of the first temperature range is generally higher than the temperature value of the second temperature range.

[0008] In some embodiments, at least two temperature control zones include a first temperature control zone, a second temperature control zone, and a third temperature control zone. The temperature of the first temperature control zone is in a first temperature range, the temperature of the second temperature control zone is in a second temperature range, and the temperature of the third temperature control zone is in a third temperature range. The temperature value of the first temperature range is higher than the temperature values ​​of the second and third temperature ranges, and the second temperature range has a segment that is higher than the third temperature range.

[0009] In some embodiments, the temperature control component includes a heat regulation section located on at least one side of the temperature control zone along a second direction and configured to heat or cool the temperature control zone; wherein the second direction is perpendicular to the first direction.

[0010] In some embodiments, the temperature control component includes multiple sets of heat adjustment parts arranged along a first direction, each set of heat adjustment parts including two heat adjustment parts arranged opposite to each other and spaced apart along a second direction, and a temperature control zone is formed between the two heat adjustment parts.

[0011] In some embodiments, the nucleic acid amplification system further includes: The second drive assembly, which is connected to multiple sets of thermal adjustment units, is configured to drive the thermal adjustment units on both sides of the temperature control zone to move along a second direction so that they move closer to each other.

[0012] In some embodiments, the temperature control assembly further includes two temperature control brackets, which are arranged opposite to each other and spaced apart along a second direction, and the temperature control brackets are configured to support the heat adjustment portion located on the same side of the temperature control zone along the second direction. The second drive assembly includes a plurality of elastic elements, with at least one elastic element connected to the side of each temperature control bracket away from the temperature control zone along the second direction. The elastic elements extend along the second direction and are configured to apply pressure along the second direction toward the temperature control zone to the connected temperature control bracket.

[0013] In some embodiments, the nucleic acid amplification system further includes: The first guide mechanism is configured to guide the movement of the heat regulation sections on both sides of the temperature control zone in the second direction.

[0014] In some embodiments, the temperature control assembly further includes two temperature control brackets, which are arranged opposite to each other and spaced apart along a second direction. The temperature control brackets are configured to support the heat adjustment portion located on the same side of the temperature control zone along the second direction. The temperature control brackets include a first mating portion. The nucleic acid amplification system also includes a main support assembly, which includes a second mating portion. A first driving assembly is configured to drive a temperature control support to move relative to the main support assembly in a first direction, so that the first mating portion moves closer to the second mating portion in the first direction. The second mating portion is configured to apply pressure to the first mating portion in a second direction away from the temperature control zone.

[0015] In some embodiments, the temperature control component further includes: Multiple first heat-conducting elements are disposed corresponding to multiple heat-regulating sections, located on the side of the heat-regulating section near the temperature control zone along the second direction, and configured to transfer the cold or heat released by the heat-regulating section to the temperature control zone; and / or Multiple second heat-conducting elements are provided corresponding to multiple heat-regulating parts and multiple first heat-conducting elements. The second heat-conducting elements are connected to the corresponding first heat-conducting elements and heat-regulating parts along a second direction to form a continuous heat transfer path.

[0016] In some embodiments, the temperature control component further includes: Multiple heat insulation elements are provided corresponding to multiple heat regulation sections, with the heat insulation elements located on the side of the corresponding heat regulation section away from the temperature control zone along the second direction.

[0017] In some embodiments, the temperature control component further includes: The first detection device is configured to detect the temperature value of the thermal conditioning unit; and The control device is signal-connected to the first detection device and the thermal regulation unit, and is configured to control the operating state of the thermal regulation unit according to the detection result of the first detection device.

[0018] In some embodiments, the temperature control component further includes: The first circuit board is electrically connected to the thermal regulation unit and the first detection device, and is signal-connected to the control device. It is configured to transmit the detection signal of the first detection device to the control device and the control signal of the control device to the thermal regulation unit.

[0019] In some embodiments, the temperature control component further includes: The second circuit board is perpendicular to the first circuit board. The second circuit board is electrically connected to the first circuit board via pin headers and is also connected to the control device for signal transmission.

[0020] In some embodiments, the nucleic acid amplification system further includes: The heat dissipation device is configured to dissipate heat from the temperature control zone when the temperature in the temperature control zone exceeds a preset threshold.

[0021] In some embodiments, the nucleic acid amplification system further includes: The second guiding mechanism is configured to guide the relative movement of the amplification section and the temperature control assembly along the first direction.

[0022] In some embodiments, the nucleic acid amplification system further includes: The second detection device is configured to detect the relative positions of the amplification unit and the temperature control component along the first direction.

[0023] In some embodiments, the nucleic acid amplification system further includes: An optical fiber is configured to emit fluorescence into the amplification cavity; and The third detection device is used to receive the fluorescence signal inside the amplification chamber.

[0024] In some embodiments, the temperature control component includes a thermal adjustment section located on a first side of the temperature control zone along a second direction and movable along the first direction, the thermal adjustment section being configured to heat or cool the temperature control zone. The nucleic acid amplification system also includes a main support assembly, which is fixed in a first direction and located on the second side of the temperature control zone in a second direction, with an optical fiber connected to the main support assembly. The second direction is perpendicular to the first direction.

[0025] In some embodiments, the temperature control assembly further includes two temperature control brackets, which are arranged opposite to each other and spaced apart along a second direction. One temperature control bracket is configured to support a heat adjustment section located on the same side of the temperature control zone along the second direction, and the other temperature control bracket is connected to the main bracket assembly and configured to support an optical fiber. The temperature control bracket connected to the main support assembly includes multiple first grooves; and / or The amplification section includes a plurality of second grooves located on the side of the amplification section facing the main support assembly along a second direction.

[0026] In some embodiments, the temperature control component includes: The thermal regulation unit is configured to heat or cool the temperature-controlled area; and The first heat-conducting element is provided corresponding to the heat regulation section and is located on the side of the heat regulation section close to the temperature control zone along the second direction, the second direction being perpendicular to the first direction; The light outlet is located on the first heat-conducting component and faces the amplification cavity. Fluorescence shines into the amplification cavity through the light outlet.

[0027] In some embodiments, the first thermal conductive element includes a limiting portion located on the side of the first thermal conductive element away from the temperature control zone along the second direction, and is configured to restrict the movement of the optical fiber toward the temperature control zone along the second direction.

[0028] In some embodiments, the amplification unit includes multiple amplification cavities, which are arranged in layers, and the light outlet is provided in a one-to-one correspondence with the amplification cavity in the same layer.

[0029] A second aspect of this disclosure provides a nucleic acid detection system, including the embodiments described above.

[0030] The third aspect of this disclosure provides a nucleic acid amplification method based on the nucleic acid amplification system of the above embodiments.

[0031] In some embodiments, the temperature control zone has a target temperature and / or an overshoot temperature, and the nucleic acid amplification method includes: Adjust the temperature of the temperature control zone to the target temperature and / or overshoot temperature; and The amplification unit is controlled to cycle between at least two temperature-controlled zones to achieve nucleic acid amplification.

[0032] In some embodiments, at least two temperature control zones include a first temperature control zone and a second temperature control zone. The target temperature and / or overshoot temperature of the first temperature control zone is within a first temperature range, and the target temperature and / or overshoot temperature of the second temperature control zone is within a second temperature range. The temperature value of the first temperature range is higher than the temperature value of the second temperature range. Controlling the amplification unit to cyclically switch between at least two temperature control zones includes: The control unit cycles between the first and second temperature control zones.

[0033] In some embodiments, at least two temperature control zones include a first temperature control zone, a second temperature control zone, and a third temperature control zone. The target temperature and / or overshoot temperature of the first temperature control zone is within a first temperature range, the target temperature and / or overshoot temperature of the second temperature control zone is within a second temperature range, and the target temperature and / or overshoot temperature of the third temperature control zone is within a third temperature range. The temperature value of the first temperature range is higher than the temperature values ​​of the second and third temperature ranges. The second temperature range has a segment that is higher than the third temperature range. Controlling the amplification unit to cyclically switch between at least two temperature control zones includes: The control amplification unit cycles through the first, second, and third temperature control zones.

[0034] In some embodiments, pre-adjusting the temperature of at least two temperature control zones includes: Adjust the temperatures of the first, second, and third temperature control zones to the target temperature; Controlling the amplification unit to cycle between at least two temperature-controlled zones includes: The control unit sequentially switches between the first temperature control zone, the third temperature control zone, and the second temperature control zone.

[0035] In some embodiments, pre-adjusting the temperature of at least two temperature control zones includes: Adjust the temperatures of the first and third temperature control zones to the target temperatures; and Set the second temperature control zone to overshoot temperature; Controlling the amplification unit to cycle between at least two temperature-controlled zones includes: During each cycle switching process, the control amplification unit switches sequentially from the first temperature control zone to the second temperature control zone, then to the third temperature control zone, and finally back to the first temperature control zone.

[0036] In some embodiments, controlling the amplification unit to cycle between at least two temperature control zones includes: During each cycle switching process, the control amplification unit switches sequentially from the first temperature control zone to the third temperature control zone, then to the second temperature control zone, and finally back to the first temperature control zone. Pre-setting the temperature of at least two temperature control zones includes: When the temperature control zone is only at the target temperature, adjust the temperature of the temperature control zone to the target temperature; and When the temperature control zone has the target temperature and the overshoot temperature, the temperature of the temperature control zone is first adjusted to the overshoot temperature, and then adjusted to the target temperature.

[0037] This disclosure provides a fourth aspect of a nucleic acid detection method based on the nucleic acid amplification system of the above embodiments, comprising: Emit fluorescence into the amplification chamber; and Detect the fluorescence signal within the amplification chamber.

[0038] In some embodiments, the amplification unit includes multiple amplification cavities arranged in layers, emitting fluorescence into each amplification cavity layer by layer and detecting the fluorescence signal within the amplification cavity.

[0039] Based on the nucleic acid amplification system provided in this disclosure, a temperature control component is configured to independently adjust at least two temperature control zones arranged along a first direction to form a temperature-cyclical environment required for nucleic acid amplification. A first driving component is used to drive the amplification section and / or the temperature control component, causing the amplification section and the temperature control component to move relative to each other along the first direction, thereby cyclically switching the amplification chamber between at least two of the temperature control zones. The temperature of at least some of the temperature control zones in this nucleic acid amplification system is adjustable, thereby enabling the adjustment of the temperature difference between each temperature control zone to meet the temperature conditions required for PCR cycling in one go. This reduces the waiting time between reactions for temperature zone adjustment, which is beneficial for accelerating the PCR cycling process, reducing the attenuation of heat transfer efficiency of amplification reagents, and extending the actual holding time of each stage of the PCR reaction, thus improving nucleic acid amplification efficiency. Furthermore, using the temperature control component to achieve independent temperature adjustment within each temperature control zone is beneficial for precise temperature control within each zone, thereby improving the temperature control accuracy of the nucleic acid amplification system.

[0040] The nucleic acid detection system disclosed herein includes the above embodiments, and thus has the advantages of the above embodiments of this disclosure.

[0041] The nucleic acid amplification method disclosed herein is based on the nucleic acid amplification system of the above embodiments, and thus has the advantages of the nucleic acid amplification system of the above embodiments of this disclosure.

[0042] The nucleic acid detection method disclosed herein is based on the nucleic acid amplification system of the above embodiments, and thus has the advantages of the nucleic acid amplification system of the above embodiments of this disclosure.

[0043] Other features and advantages of this disclosure will become clear from the following detailed description of exemplary embodiments with reference to the accompanying drawings. Attached Figure Description

[0044] The accompanying drawings, which are included to provide a further understanding of this disclosure and form part of this application, illustrate exemplary embodiments of this disclosure and are used to explain this disclosure, but do not constitute an undue limitation of this disclosure. In the drawings: Figure 1 This is a schematic diagram of the structure of a nucleic acid amplification system according to some embodiments of this disclosure.

[0045] Figure 2 This is a schematic diagram of the structure of the temperature control component of a nucleic acid amplification system in some embodiments of this disclosure in an expanded state.

[0046] Figure 3 for Figure 2 The diagram shows a front perspective view of the nucleic acid amplification system, which reveals the location of the second ligation region.

[0047] Figure 4 for Figure 3The diagram shown is a partial enlarged view of the nucleic acid amplification system, which illustrates the first and second coordinating parts.

[0048] Figure 5 This is a schematic diagram of the temperature control component of a nucleic acid amplification system according to some embodiments of the present disclosure in a closed state.

[0049] Figure 6 for Figure 5 The diagram shows a front perspective view of the nucleic acid amplification system, which reveals the location of the second ligation region.

[0050] Figure 7 for Figure 5 The diagram shown is an exploded view of the nucleic acid amplification system.

[0051] Figure 8 for Figure 7 The exploded view of the temperature control component of the nucleic acid amplification system located on the first side of the second direction in the temperature control zone.

[0052] Figure 9 for Figure 7 The exploded view of the temperature control component of the nucleic acid amplification system located on the second side of the second direction in the temperature control zone.

[0053] Figure 10 This is a cross-sectional view of an optical fiber mounted on a first heat-conducting element according to some embodiments of this disclosure.

[0054] Figure 11 This is a schematic diagram of the structure of a first heat-conducting element according to some embodiments of this disclosure.

[0055] Figure 12 This is a side view of a microfluidic card cartridge according to some embodiments of this disclosure.

[0056] Figure 13 This is a schematic diagram of the structure of a temperature control component at a first detection position, according to some embodiments of this disclosure.

[0057] Figure 14 This is a schematic diagram of the structure of a temperature control component at a second detection position, representing some embodiments of this disclosure.

[0058] Figure 15 This is a schematic diagram of the structure of a nucleic acid amplification system according to some embodiments of this disclosure.

[0059] Figure 16 This is a schematic diagram of the structure of a nucleic acid amplification system according to some embodiments of this disclosure.

[0060] Figure 17 for Figure 16 A top view of a portion of the structure of the nucleic acid amplification system shown.

[0061] Figure 18This is a schematic diagram of the structure of the amplification section in some embodiments of this disclosure.

[0062] Figure 19 This is a temperature diagram of nucleic acid amplification reactions in related technologies, showing the temperature conditions for the denaturation, annealing, and extension phases.

[0063] Explanation of reference numerals in the attached figures: 10. Mating part bracket; 101. Second mating part; 102. Light baffle; 103. Light opening plate; 11. Support plate; 111. Back plate; 12. Guide rod; 13. Guide rail; 14. Support leg; 15. Annular frame; 16. First mating part; 161. Third wall; 17. Drive motor; 170. Fixing plate; 18. Mounting plate; 19. Guide block; 20. Flange; 2. Temperature control assembly; 21. First heat conduction component; 211. Limiting part; 212. Light outlet; 23. Heat adjustment part; 24. Heat insulation component; 27. First detection device; 200. Temperature control bracket; 201. First circuit board; 202. Cable bundle plate; 203. Second circuit board; 3. Heat dissipation device; 4. Heat dissipation device mounting plate; 5. Limiting sheet metal; 6. Linear bearing; 7. Elastic element; 8. Optical fiber; 9. Microfluidic cartridge; 90. Amplification chamber; 900. Amplification channel; 91. Microfluidic chip; 911. Second groove; 92. Cartridge body; x, the first direction; y, the second direction. Detailed Implementation

[0064] The technical solutions of the embodiments of this disclosure will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this disclosure, and not all embodiments. The following description of at least one exemplary embodiment is merely illustrative and is in no way intended to limit this disclosure or its application or use. All other embodiments obtained by those skilled in the art based on the embodiments of this disclosure without creative effort are within the scope of protection of this disclosure.

[0065] Unless otherwise specifically stated, the relative arrangement, numerical expressions, and values ​​of the components and steps set forth in these embodiments do not limit the scope of this disclosure. It should also be understood that, for ease of description, the dimensions of the various parts shown in the drawings are not drawn to actual scale. Techniques, methods, and devices known to those skilled in the art may not be discussed in detail, but where appropriate, such techniques, methods, and devices should be considered part of the specification. In all examples shown and discussed herein, any specific values ​​should be interpreted as merely exemplary and not as limitations. Therefore, other examples of exemplary embodiments may have different values. It should be noted that similar reference numerals and letters in the following drawings denote similar items; therefore, once an item is defined in one drawing, it need not be further discussed in subsequent drawings.

[0066] In the description of this disclosure, it should be understood that the use of terms such as "first" and "second" to define components is merely for the purpose of distinguishing the corresponding components. Unless otherwise stated, the above terms have no special meaning and therefore should not be construed as limiting the scope of protection of this disclosure.

[0067] In the description of this disclosure, it should be understood that the orientation or positional relationship indicated by directional terms such as "front, back, up, down, left, right", "horizontal, vertical, vertical, horizontal" and "top, bottom" are generally based on the orientation or positional relationship shown in the accompanying drawings, and are only for the convenience of describing this disclosure and simplifying the description. Unless otherwise stated, these directional terms do not indicate or imply that the device or element referred to must have a specific orientation or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation on the scope of protection of this disclosure; the directional terms "inner" and "outer" refer to the inner and outer contours relative to the outline of each component itself.

[0068] In the following description, "front" and "back" correspond to perpendicular to Figure 3 and Figure 6 The orientation of the paper; "left" and "right" refer to the left and right directions when facing forward; the so-called first direction x refers to... Figures 1 to 3 , Figure 5 and Figure 6 The direction extending along the x-coordinate; the so-called second direction y refers to Figures 1 to 3 , Figure 5 and Figure 6 The direction in which the y-coordinate extends.

[0069] PCR reactions mainly consist of pre-denaturation, denaturation, annealing, and extension phases. The denaturation-annealing-extension phase is cyclical, with one run constituting one PCR cycle. Typically, a single nucleic acid amplification cycle comprises 30-50 PCR cycles. Theoretically, the amount of nucleic acid molecules doubles after each PCR cycle. Throughout the PCR amplification process, the amount of DNA template increases exponentially with the number of cycles. For pathogens whose genetic material is RNA, a reverse transcription phase is added before pre-denaturation to synthesize complementary DNA using RNA as a template for subsequent nucleic acid amplification. Each stage of the PCR reaction requires specific temperature conditions, and each stage also requires a certain temperature holding time to ensure the complete execution of the corresponding biological reaction.

[0070] The main functions of the pre-denaturation stage are: to open the tertiary and secondary structures of the target nucleic acid chains, separating the double-stranded DNA template into single strands, and improving the efficiency of subsequent PCR amplification. When some PCR reactions use hot-start DNA polymerase, the enzyme is inactive at low temperatures. The high temperature of the pre-denaturation stage can separate the protective molecules from the enzyme, thereby activating its activity. The high temperature of pre-denaturation can also remove specific bindings to some extent. The temperature of the pre-denaturation stage is usually around 95℃. Too high a temperature will affect enzyme activity, while too low a temperature will prevent the nucleic acid chains from fully opening and enzyme activity from being fully activated, thus affecting detection sensitivity.

[0071] The main function of the denaturation stage is to unwind double-stranded DNA into single strands, facilitating subsequent binding with primers. For example... Figure 19 As shown, the denaturation stage temperature is usually around 95℃. Too high a temperature will affect enzyme activity, while too low a temperature may lead to incomplete denaturation.

[0072] like Figure 19 As shown, the main function of the annealing stage is to allow the primers and template single strands to bind in a complementary base pairing manner after the denaturation stage, when the temperature drops to around 55°C. Excessively high temperatures hinder primer-template binding, while excessively low temperatures lead to non-specific amplification.

[0073] The main function of the extension phase is that after the primer binds to the template, the free nucleotides, under the action of DNA polymerase, will bind to the single-stranded DNA according to the base pairing principle, extending from the primer in the previous stage, synthesizing a complementary strand along the direction from phosphate to pentose, and generating double-stranded DNA. The extension phase begins when the primer binds to the template, as shown below. Figure 19As shown, the optimal reaction temperature for DNA polymerase is around 72℃, but it also exhibits activity during the transition from the annealing stage to 72℃. Therefore, when the target nucleic acid fragment to be amplified is short (100~300bp), a two-step PCR method can be used if PCR technology is applied to clinical diagnosis. Studies have shown that the extension phase can occur during the transition between annealing and denaturation temperatures, and additional holding time is not required when the DNA amplification target is short.

[0074] like Figures 1 to 14 As shown, this disclosure provides a nucleic acid amplification system. The nucleic acid amplification system includes: The microfluidic cartridge 9 includes an amplification section having an amplification cavity 90 for containing amplification reagents; At least two temperature control zones are arranged along a first direction x, and the temperature control zones are configured to accommodate the amplification section, with at least a portion of the temperature control zones having adjustable temperatures; Temperature control component 2 is configured to independently adjust the temperature of each temperature control zone, so that the temperature of each temperature control zone is different; and A first driving component, drivenly connected to the amplification section and / or the temperature control component 2, is configured to move the amplification section and the temperature control component 2 relative to each other along a first direction x, so that the amplification chamber 90 cycles between at least two temperature control zones.

[0075] like Figure 12 As shown, for example, the microfluidic cartridge 9 includes a microfluidic chip 91 and a cartridge body 92. The cartridge body 92 has an opening, and a first end of the microfluidic chip 91 is inserted into the opening. A second end of the microfluidic chip 91 extends away from the cartridge body 92. The microfluidic chip 91 includes an amplification section, which includes an amplification channel 900 and an amplification cavity 90. The amplification channel 900 connects the cartridge body 92 and the amplification cavity 90, so that the amplification reagent in the cartridge body 92 can flow into the amplification cavity 90.

[0076] For example, at least a portion of the box body 92 may be a cuboid, or it may be a cylindrical structure such as a prism or cylinder, in order to improve the adaptability of the microfluidic card box 9 to the installation space.

[0077] For example, the first direction x can be vertical or horizontal, which helps reduce the contact area between temperature zones, thus avoiding thermal crosstalk and reducing the footprint of the nucleic acid amplification system. Alternatively, the first direction x can be circumferential, allowing each temperature control zone to be arranged in a ring along the circumference according to the temperature cycle sequence required for PCR amplification. This ensures that the spatial arrangement of the temperature control zones matches the temporal requirements of the temperature cycle, allowing the amplification unit to continuously and sequentially pass through each temperature control zone. Each complete cycle of PCR temperature cycling is achieved automatically without reciprocating motion or reversing direction, thus facilitating a continuous and smooth temperature cycling process.

[0078] Based on the nucleic acid amplification system provided in this embodiment, a temperature control component 2 is configured to independently adjust at least two temperature control zones arranged along a first direction x, thereby forming a temperature cycling environment required for nucleic acid amplification. A first driving component is used to drive the amplification section and / or the temperature control component 2, causing the amplification section and the temperature control component 2 to move relative to each other along the first direction x, so that the amplification chamber 90 cycles between at least two of the temperature control zones. The temperature of at least some of the temperature control zones in this nucleic acid amplification system is adjustable, thereby enabling the adjustment of the temperature difference between each temperature control zone to meet the temperature conditions required for PCR cycling in one go. This reduces the waiting time between reactions for the temperature control zones to adjust their temperatures, which helps to accelerate the PCR cycling process, reduce the attenuation of heat transfer efficiency of the amplification reagents, and extend the actual holding time of each stage of the PCR reaction, thus improving nucleic acid amplification efficiency. Furthermore, using the temperature control component 2 to achieve independent temperature adjustment within each temperature control zone facilitates precise temperature control within each zone, thereby improving the temperature control accuracy of the nucleic acid amplification system.

[0079] In some embodiments, the first driving component is driven to the temperature control component 2 to drive the temperature control component 2 to reciprocate along the first direction x. In other embodiments, the first driving component is driven to the amplification unit to drive the amplification unit to reciprocate along the first direction x.

[0080] In some embodiments, each temperature control zone has a target temperature and an overshoot temperature. The temperature change rate of the amplification section in the temperature control zone at the overshoot temperature is higher than the temperature change rate of the amplification section in the temperature control zone at the target temperature, thereby improving the amplification efficiency of the nucleic acid amplification system. In some embodiments, at least two temperature control zones include a first temperature control zone and a second temperature control zone. The temperature of the first temperature control zone is within a first temperature range, and the temperature of the second temperature control zone is within a second temperature range. The temperature values ​​of the first temperature range are generally higher than the temperature values ​​of the second temperature range.

[0081] For example, the first temperature range is 93℃-98℃. The second temperature range is 50℃-65℃.

[0082] For example, in one PCR cycle, the microfluidic chip 91 directly switches from the first temperature control zone to the second temperature control zone, and then switches back from the second temperature control zone to the first temperature control zone.

[0083] For example, the first temperature control zone and the second temperature control zone can be arranged along the height direction, and the first temperature control zone can be located below the second temperature control zone.

[0084] Setting up a first temperature control zone and a second temperature control zone can shorten the relative movement distance of the amplification section in each PCR cycle, thereby reducing the non-reaction time in each PCR cycle and improving amplification efficiency.

[0085] In some embodiments, the first temperature control region may only have a first target temperature, for example, the first target temperature is set to 95°C. In other embodiments, the first temperature control region may simultaneously have a first target temperature and a first overshoot temperature, where the first target temperature is lower than the first overshoot temperature; for example, the first target temperature is set to 95°C and the first overshoot temperature is set to 98°C. By setting the first overshoot temperature, the amplification section located within the first temperature control region is rapidly heated, thereby improving amplification efficiency.

[0086] In some embodiments, the second temperature control zone may have only a second target temperature, for example, the second target temperature is set to 55°C. In other embodiments, the second temperature control zone may have both a second target temperature and a second overshoot temperature, the second overshoot temperature being lower than the second target temperature, for example, the second target temperature is set to 55°C and the second overshoot temperature is set to 52°C.

[0087] For example, in rapid detection of respiratory viruses such as COVID-19, the first target temperature of the first temperature control zone is set to 95℃, and the first overshoot temperature is set to 98℃. The second target temperature of the second temperature control zone is set to 58℃, and the second overshoot temperature is set to 53℃. In one PCR cycle, the first temperature control zone is first adjusted to the first overshoot temperature. After the amplification section of the microfluidic chip 91 remains in the first temperature control zone for a preset time (so that the temperature of the amplification reagent is close to the first target temperature), the temperature of the first temperature control zone is then adjusted back to the first target temperature. The temperature adjustment method for the second temperature control zone is the same as that for the first temperature control zone, and will not be described again here.

[0088] In some embodiments, at least two temperature control zones include a first temperature control zone, a second temperature control zone, and a third temperature control zone. The temperature of the first temperature control zone is in a first temperature range, the temperature of the second temperature control zone is in a second temperature range, and the temperature of the third temperature control zone is in a third temperature range. The temperature value of the first temperature range is higher than the temperature values ​​of the second and third temperature ranges, and the second temperature range has a segment that is higher than the third temperature range.

[0089] For example, the range of the first temperature range can be 93℃-98℃, the range of the second temperature range can be 50℃-75℃, and the range of the third temperature range can be 50℃-65℃.

[0090] For example, such as Figure 7 As shown, the first temperature control zone, the second temperature control zone, and the third temperature control zone can be arranged along the height direction, with the first temperature control zone at the bottom, the third temperature control zone at the top, and the second temperature control zone located between the first and third temperature control zones along the height direction.

[0091] For detection scenarios involving long DNA fragments, setting up a first, second, and third temperature control zone can simultaneously meet the temperature requirements of the denaturation, annealing, and extension phases in the PCR reaction. This allows the temperature control component 2 to provide independent and precisely controllable reaction temperatures for each of these phases, improving amplification efficiency. Furthermore, the annealing phase in the PCR reaction needs to be carried out within a temperature range of 50℃-65℃ to ensure the "specificity" of primer binding to the template DNA. Therefore, setting up multiple temperature control zones and independently adjusting the temperature of each zone helps reduce temperature fluctuations and facilitates stable annealing of the PCR reaction at low temperatures. This makes the nucleic acid amplification system suitable for clinical nucleic acid detection scenarios where high accuracy and reliability of reaction temperatures are required.

[0092] For detection scenarios involving short DNA fragments, one of the temperature control zones can be kept at an overshoot temperature below the annealing temperature. This allows the amplification section to cool down rapidly while it remains in that temperature control zone, thereby improving detection efficiency.

[0093] In some embodiments, during a single PCR cycle, the microfluidic chip 91 can first perform a denaturation reaction in a first temperature-controlled zone, then switch to a third temperature-controlled zone for an annealing reaction, then switch to a second temperature-controlled zone for an extension reaction, and finally switch back to the first temperature-controlled zone for the next cycle. Notably, the microfluidic chip 91 does not need to remain in the second temperature-controlled zone during the switching process from the first to the third temperature-controlled zone.

[0094] In some embodiments, during a PCR cycle, the microfluidic chip 91 can first perform a denaturation reaction in a first temperature control zone, then switch to a second temperature control zone for rapid cooling, then switch to a third temperature control zone for annealing, and finally switch back to the first temperature control zone for the next cycle.

[0095] In some embodiments, the temperature setting method of the first temperature control zone for PCR reaction using three temperature control zones can be the same as that of the first temperature control zone for PCR reaction using two temperature control zones, and will not be described again here.

[0096] In some embodiments, the second temperature control zone may only have a second target temperature, for example, the second target temperature is set to 72°C to meet the extension temperature conditions in the PCR reaction.

[0097] In some embodiments, the second temperature control zone may have only a second overshoot temperature, for example, the second overshoot temperature is set to 50°C, so that the amplification section cools down rapidly before entering the third temperature control zone.

[0098] In some embodiments, the second temperature control zone may simultaneously have a second target temperature and a fourth overshoot temperature. For the detection of long DNA fragments, the second target temperature can be set as the temperature for the extension reaction, and the fourth overshoot temperature can be used to rapidly heat the amplification section, thereby shortening the time required for the amplification section to rise from the annealing temperature to the extension temperature. For example, the fourth overshoot temperature is 75°C.

[0099] In some embodiments, the third temperature control zone may only have a third target temperature, for example, the third target temperature is set to 55°C. In other embodiments, the third temperature control zone may simultaneously have a third target temperature and a third overshoot temperature, where the third target temperature is higher than the third overshoot temperature; for example, the third target temperature is 55°C and the third overshoot temperature is 50°C. By setting the third overshoot temperature, the amplification section located in the third temperature control zone can be rapidly cooled, thereby extending the time the amplification section spends in a low-temperature environment, which is beneficial to the stability of the PCR reaction.

[0100] In some embodiments, any two of the first, second, and third temperature control zones can be selected as the temperature control zones for the denaturation reaction and the annealing reaction, respectively (without the need for an extension reaction). For example, the temperature range of the first temperature control zone can be set to 93℃-98℃, and the range of the second temperature control zone can be set to 50℃-65℃. As another example, the temperature range of the second temperature control zone can be set to 93℃-98℃, and the range of the third temperature control zone can be set to 50℃-65℃.

[0101] like Figures 7 to 9 As shown, in some embodiments, the temperature control component 2 includes a heat adjustment section 23, which is located on at least one side of the temperature control zone along the second direction y and is configured to heat or cool the temperature control zone; wherein the second direction y is perpendicular to the first direction x.

[0102] For example, the thermal regulation unit 23 can be a heating element or a semiconductor cooling element.

[0103] The heat regulation unit 23 is disposed on at least one side of the temperature control zone along the second direction y, such that the heat conduction direction between the heat regulation unit 23 and the amplification unit is perpendicular to the movement direction of the amplification unit relative to the temperature control component 2. This vertical arrangement can shorten the heat conduction path, improve the uniformity and efficiency of heat conduction, and reduce the heat dissipation to areas outside the temperature control zone, thus ensuring rapid and stable temperature regulation of the amplification unit.

[0104] In some embodiments (not shown), the thermal adjustment section 23 may be arranged around the temperature control zone on three sides with the first direction x as the axial direction, and form an opening through which the microfluidic chip 91 can extend into the temperature control zone. This arrangement helps to increase the heat conduction area, thereby improving the temperature adjustment speed.

[0105] like Figure 2 , Figure 3 , Figures 5 to 9 As shown, in some embodiments, the temperature control component 2 includes multiple sets of heat adjustment parts 23 arranged along the first direction x. Each set of heat adjustment parts 23 includes two heat adjustment parts 23 arranged opposite to each other and spaced apart along the second direction y, and a temperature control zone is formed between the two heat adjustment parts 23.

[0106] For example, the temperature control component 2 may include two sets of heat adjustment parts 23 arranged along the first direction x to form a first temperature control zone and a second temperature control zone, respectively. As another example, such as... Figure 7 As shown, the temperature control component 2 may also include three sets of heat adjustment parts 23 arranged along the first direction x, so as to form a first temperature control zone, a second temperature control zone and a third temperature control zone respectively.

[0107] A temperature control zone is formed between the two thermal regulation units 23. Each temperature control zone is regulated by two thermal regulation units 23, which can increase the heat exchange area, improve the heat conduction efficiency and heat input and output capacity, thereby improving the temperature regulation speed and response rate of the temperature control zone, enabling the amplification unit to quickly reach the target temperature, and thus improving the amplification reaction efficiency.

[0108] like Figure 5 and Figure 6 As shown, in some embodiments, the nucleic acid amplification system further includes: The second drive assembly, which is driven connected to multiple sets of thermal adjustment units 23, is configured to drive the thermal adjustment units 23 on both sides of the temperature control zone to move along the second direction y to move closer to each other.

[0109] The second driving component drives the thermal adjustment sections 23 on both sides of the temperature control zone to move closer to each other along the second direction y. This allows the thermal adjustment sections 23 on both sides to converge inwards and approach the microfluidic chip 91 along the second direction y, which helps to shorten the distance between the thermal adjustment sections 23 and the amplification section, thereby improving heat exchange efficiency. In addition, the second driving component ensures that the distance between the thermal adjustment sections 23 and the amplification section on both sides of the temperature control zone is consistent, which helps to ensure uniform heating on both sides of the amplification section.

[0110] like Figures 1 to 3 , Figures 5 to 7 As shown, in some embodiments, the temperature control assembly 2 further includes two temperature control brackets 200, which are arranged opposite to each other and spaced apart along the second direction y. The temperature control brackets 200 are configured to support the heat adjustment part 23 located on the same side of the temperature control zone along the second direction y. The second drive assembly includes a plurality of elastic elements 7, with at least one elastic element 7 connected to the side of each temperature control bracket 200 away from the temperature control zone along the second direction y. The elastic element 7 extends along the second direction y and is configured to apply pressure to the connected temperature control bracket 200 along the second direction y toward the temperature control zone.

[0111] For example, two elastic elements 7 can be connected to the side of each temperature control bracket 200 away from the temperature control zone along the second direction y, and these two elastic elements 7 are spaced apart along the first direction x.

[0112] For example, the elastic element 7 is a spring, etc.

[0113] The temperature control bracket 200 integrates the heat regulation units 23 located on the same side along the second direction y. The elastic element 7 drives the temperature control bracket 200 to drive each heat regulation unit 23, enabling synchronous movement of the heat regulation units 23 on the same side along the second direction y. This simplifies the drive structure and improves drive efficiency and stability. Furthermore, the elastic element 7 has a simple structure, facilitating flexible setting of its installation position and elastic force. The elastic element 7 provides continuous elastic clamping force, ensuring smooth movement of the amplification unit along the first direction x while shortening the distance between the heat regulation unit 23 and the amplification unit, further improving the stability and reliability of temperature transfer.

[0114] In some embodiments (not shown), the second driving component may also include a motor, the output of which is driven and connected to the temperature control bracket 200. The motor is configured to drive the temperature control bracket 200 to reciprocate along the second direction y, thereby causing the heat regulation section 23, located on the same side along the second direction y, to synchronously approach the amplification section, achieving stable heating of the amplification section. By driving the motor, the moving position and clamping force of the temperature control bracket 200 can be precisely controlled, improving the accuracy of the distance between the heat regulation section 23 and the amplification section and the heating stability, enhancing temperature transfer efficiency, and simultaneously improving the automation level and operational reliability of the nucleic acid amplification system.

[0115] In some embodiments (not shown), the second drive assembly may also include two magnets, each corresponding to one of the two temperature control brackets 200. The magnetic force of the two magnets can drive the two temperature control brackets 200 to move closer to each other along the second direction y, so that the thermal adjustment section 23 is closer to the amplification section. Including two magnets in the second drive assembly simplifies the drive structure and improves installation efficiency.

[0116] like Figure 7 As shown, in some embodiments, the nucleic acid amplification system further includes: The first guide mechanism is configured to guide the movement of the heat adjustment units 23 on both sides of the temperature control zone along the second direction y.

[0117] like Figure 7As shown, for example, the first guiding mechanism includes two guide rods 12 extending along a second direction y. The temperature control bracket 200 includes two through holes spaced apart along a first direction x. The through holes extend along the second direction y. The two ends of the guide rods 12 are respectively supported in the through holes of the two temperature control brackets 200 located on the same side along the first direction x. The first guiding mechanism also includes two linear bearings 6, which are respectively disposed corresponding to the two temperature control brackets 200. The linear bearings 6 drive the temperature control brackets 200 to move along the extension direction of the guide rods 12.

[0118] In other embodiments (not shown), the first guiding mechanism may include a cooperating guide rail and a slider. The guide rail is disposed on the main support assembly and extends in a second direction. The main support assembly is configured to support the temperature control component 2. The slider may be disposed on the temperature control bracket 200, thereby allowing the temperature control bracket 200 to move along the extension direction of the guide rail through the cooperation of the guide rail and the slider.

[0119] Setting up a first guiding mechanism can improve the stability and positional accuracy of the movement of the two temperature control brackets 200 along the second direction y, ensure the distance and positional accuracy between the thermal regulation section 23 and the amplification section, and thus help improve the heat transfer efficiency.

[0120] like Figures 2 to 7 As shown, in some embodiments, the temperature control bracket 200 includes a first mating portion 16; The nucleic acid amplification system also includes a main support assembly, which includes a second mating part 101. A first driving assembly is configured to drive a temperature control support 200 to move relative to the main support assembly along a first direction x, so that the first mating part 16 moves closer to the second mating part 101 along the first direction x. The second mating part 101 is configured to apply pressure to the first mating part 16 along a second direction y away from the temperature control zone.

[0121] By cooperating with the first mating part 16 and the second mating part 101, the two temperature control brackets 200 move away from each other along the second direction y, thereby forming a space that allows the microfluidic chip 91 to be inserted into each temperature control zone. This helps to avoid interference, scratching or jamming between the microfluidic chip 91 and the temperature control component 2 during insertion and removal, and improves the smoothness and convenience of the operation process.

[0122] like Figures 2 to 7As shown, for example, the temperature control bracket 200 also includes bakelite configured to support the heat adjustment section 23 located on the same side of the temperature control zone along the second direction y. The first mating section 16 includes a first end and a second end. The second end is connected to the side of the bakelite along the second direction y away from the plurality of temperature control zones. The first end is a free end, and a first mating segment of the first mating section 16 where the first end is located extends along the first direction x. The first mating segment includes a first wall and a second wall that are opposite and spaced apart along the second direction y, the second wall being closer to each temperature control zone along the second direction y than the first wall. The end wall of the first end connects between the first wall and the second wall, and the end wall is an inclined wall or an arc-shaped wall.

[0123] like Figure 3 , Figure 4 and Figure 6 As shown, for example, the second mating part 101 is a rotating member, and the rotating shaft extends along a third direction. The third direction is perpendicular to the first direction x and the second direction y. The second mating part 101 can be, for example, a bearing.

[0124] like Figures 2 to 6 As shown, in some embodiments, the temperature control component 2 has a closed state and an open state. A first drive component is configured to drive bakelite to move along a first direction x, causing the temperature control component 2 to switch between the closed and open states. In the closed state, the first mating part 16 and the second mating part 101 are separated from each other, and the second drive component applies pressure along the first direction x to the two temperature control supports 200 via the elastic element 7, causing the two temperature control supports 200 to move closer together, thereby enabling the thermal adjustment part 23 to approach the microfluidic chip 91. Figures 2 to 4 As shown, in some embodiments, the first mating part 16 and the second mating part 101 are gradually brought closer together by the first driving component, and the second mating part 101 comes into contact with the third wall 161, so that the temperature control component 2 switches from a closed state to an open state. The second mating part 101 moves along the third wall 161 from away from the second wall to closer to the second wall, and applies pressure to the first mating part 16 in a first direction x toward the direction away from each temperature control zone, so as to drive the two bakelite pieces away from each other in the first direction x. Finally, the second mating part 101 moves to the side of the second wall in a second direction y that is closer to each temperature control zone, at which point the temperature control component 2 switches to the open state.

[0125] like Figure 1 and Figure 7As shown, in some embodiments, the main support assembly includes two legs 14, an annular frame 15, two mating brackets 10, two second mating parts 101, and a support plate 11. The two legs 14 are arranged opposite each other and spaced apart along a second direction y. The annular frame 15 is connected between the two legs 14 and is perpendicular to a third direction. The two mating brackets 10 are arranged one-to-one with the two legs 14, and are connected to the rear top of the corresponding leg 14. The two second mating parts 101 are arranged one-to-one with the two mating brackets 10, and are connected to the front of the corresponding mating bracket 10. The support plate 11 is supported on the annular bracket 15 and is perpendicular to a first direction x. Two temperature control brackets 200 are connected to the top of the support plate 11.

[0126] like Figures 7 to 9 As shown, in some embodiments, the temperature control component 2 further includes: Multiple first heat-conducting elements 21 are disposed corresponding to multiple heat-regulating sections 23, located on the side of the heat-regulating section 23 along the second direction y near the temperature control zone, and are configured to transfer the cold or heat released by the heat-regulating section 23 to the temperature control zone; and / or Multiple second heat-conducting elements are provided corresponding to multiple heat-regulating parts 23 and multiple first heat-conducting elements 21. The second heat-conducting elements are connected to the corresponding first heat-conducting elements 21 and heat-regulating parts 23 along the second direction y to form a continuous heat transfer path.

[0127] For example, the first heat-conducting component 21 can be a heat-conducting block. The second heat-conducting component can be made of materials such as thermal grease or graphite paper.

[0128] The first heat-conducting element 21 can evenly conduct the heat generated by the thermal regulation unit 23 to the corresponding temperature control zone, which helps to improve the uniformity of temperature distribution in each temperature control zone and avoid local overheating. The first heat-conducting element 21 can also increase the thermal contact area between the temperature control component 2 and the amplification unit, improve the heat conduction efficiency, and buffer heat shock, which helps to prevent the amplification unit from being burned. The second heat-conducting element can establish a continuous heat conduction path between the thermal regulation unit 23 and the first heat-conducting element 21, which helps to reduce contact thermal resistance and improve heat transfer efficiency.

[0129] It should be noted that, in the closed state, the first heat-conducting component 21 is attached to the microfluidic chip 91, and the first driving component can drive the temperature control component 2 to move relative to the microfluidic chip 91 along the first direction x.

[0130] like Figures 7 to 9 As shown, in some embodiments, the temperature control component 2 further includes: Multiple heat insulation elements 24 are provided corresponding to multiple heat adjustment parts 23, with the heat insulation elements 24 located on the side of the corresponding heat adjustment part 23 away from the temperature control zone along the second direction y.

[0131] For example, the heat insulation element 24 can be a silicone pad to buffer pressure.

[0132] The installation of the heat insulation component 24 helps to prevent damage to the heat regulation unit 23 and also helps to reduce the influence of the external environment on heat conduction.

[0133] like Figures 7 to 9 As shown, in some embodiments, the heat insulation element 24 is connected to the temperature control bracket 200. The temperature control assembly includes screws that securely connect the bakelite, the heat insulation element 24, the thermal adjustment part 23, and the first heat-conducting element 21 to the temperature control bracket 200.

[0134] like Figures 7 to 9 As shown, in some embodiments, the temperature control component 2 further includes: The first detection device 27 is configured to detect the temperature value of the thermal regulation unit 23; and The control device is signal-connected to the first detection device 27 and the thermal adjustment unit 23, and is configured to control the operating state of the thermal adjustment unit 23 according to the detection result of the first detection device 27.

[0135] For example, the first detection device 27 can be installed inside the first heat-conducting component 21. The first detection device 27 can be a temperature sensor.

[0136] The first detection device 27 detects the temperature value of the thermal regulation unit 23, and the control device controls the thermal regulation unit 23 to heat or cool the temperature control zone. This facilitates rapid temperature adjustment in each temperature control zone, ensuring that the temperature control zone can quickly and stably reach and maintain the required temperature conditions, reducing temperature fluctuations and deviations, and improving temperature control accuracy and stability. This, in turn, helps to provide accurate and reliable temperature conditions for the PCR reaction.

[0137] The control device may be implemented as a general-purpose processor, a programmable logic controller (PLC), a digital signal processor (DSP), an application-specific integrated circuit (ASIC), a field-programmable gate array (FPGA), or other programmable logic device, discrete gate or transistor logic device, discrete hardware component, or any suitable combination thereof for performing the functions described in this disclosure.

[0138] like Figures 7 to 9 As shown, in some embodiments, the temperature control component 2 further includes: The first circuit board 201 is electrically connected to the thermal regulation unit 23 and the first detection device 27, and is signal-connected to the control device. It is configured to transmit the detection signal of the first detection device 27 to the control device and the control signal of the control device to the thermal regulation unit 23.

[0139] For example, the first circuit board 201 can be connected to one side of the temperature control bracket 200 along the third direction and is arranged perpendicular to the third direction. The first detection device 27 and the thermal adjustment part 23 can be soldered onto the first circuit board 201.

[0140] The first circuit board 201 integrates the electrical connections between the thermal regulation unit 23, the first detection device 27 and the control device, reduces the use of scattered wires, simplifies the wiring structure, and thus improves the spatial integration and structural compactness of the temperature control component 2.

[0141] like Figures 7 to 9 As shown, in some embodiments, the temperature control component 2 further includes: The second circuit board 203 is perpendicular to the first circuit board 201. The second circuit board 203 is electrically connected to the first circuit board 201 through pin headers and is also connected to the control device for signal transmission.

[0142] For example, one end of the first circuit board 201 is provided with a pin header, and the first circuit board 201 is electrically connected to the second circuit board 203 through the pin header.

[0143] Since the first circuit board 201 is close to each temperature control zone and thermal regulation unit 23, the working environment temperature varies greatly and the space is compact. The control device needs to be far away from the high temperature area to ensure working stability. Therefore, a second circuit board 203 is set between the first circuit board 201 and the control device. This enables signal transfer and electrical connection between the first circuit board 201 and the control device, blocks heat conduction from the temperature control zone to the control device, and helps protect the control device from high temperature.

[0144] like Figures 7 to 9 As shown, in some embodiments, the temperature control component 2 further includes a cable management board 202, which is connected to the second circuit board 203, and a cable management space is formed between the cable management board 202 and the second circuit board 203. The cable management space is used to integrate flexible flat cables, which helps to improve spatial integration and assembly convenience.

[0145] like Figure 7 As shown, in some embodiments, the nucleic acid amplification system further includes: The heat dissipation device 3 is configured to dissipate heat from the temperature control zone when the temperature in the temperature control zone exceeds a preset threshold.

[0146] For example, heat dissipation device 3 is a fan. Figure 7As shown, the nucleic acid amplification system may include a heat dissipation device mounting plate 4, which is connected to the main support assembly, and a heat dissipation device 3 is mounted on the heat dissipation device mounting plate 4.

[0147] The heat dissipation device 3 is beneficial for rapidly cooling down each temperature control zone and also helps to prevent excessively high temperatures in each zone, ensuring temperature control accuracy and stable and reliable PCR reaction.

[0148] like Figure 7 As shown, in some embodiments, the heat dissipation device 3 is configured to dissipate heat from the temperature control zone used for the annealing reaction.

[0149] Since the temperature difference between the temperature control zone used for the annealing reaction and room temperature is small, the heat dissipation device 3, used to dissipate heat from the temperature control zone for the annealing reaction, helps to improve the cooling rate of the temperature control zone. In addition, under specific amplification conditions, the heat dissipation device 3 can achieve a complete temperature cycle process using only one temperature control zone.

[0150] like Figures 1 to 3 , Figures 5 to 7 As shown, in some embodiments, the nucleic acid amplification system further includes: The second guiding mechanism is configured to guide the relative movement of the amplification section and the temperature control component 2 along the first direction x.

[0151] like Figure 2 , Figure 3 and Figure 7 As shown, for example, the second guiding mechanism may include a guide block 19, a guide rail 13, and a mounting plate 18. The guide rail 13 extends along a first direction x and is mounted on the annular frame 15 of the main support assembly. The guide block 19 is supported on the guide rail 13 and slides in engagement with the guide rail 13. The mounting plate 18 is connected to the guide block 19 and is perpendicularly connected to the side of the bakelite along the second direction y away from each temperature control zone. The bakelite is connected above the support plate 11 of the main support assembly. The first drive assembly includes a drive motor 17 and a fixing plate 170. The drive motor 17 is connected to the fixing plate 170, and the fixing plate 170 is fixedly connected to the support plate 11, so that the drive motor 17 can drive the support plate 11 to reciprocate along the first direction x, thereby driving the temperature control assembly 2 to reciprocate along the first direction x.

[0152] like Figures 1 to 3 , Figures 5 to 7 As shown, for example, the nucleic acid amplification system can be equipped with two second guiding mechanisms, which are respectively set with two temperature control brackets 200.

[0153] The second guide mechanism enables the mounting plate 18 to reciprocate along the first direction x, thereby driving the temperature control component 2 to reciprocate along the first direction x. The second guide mechanism is beneficial for providing stable guidance for the movement of the temperature control component 2, improving the smoothness and positional accuracy of the movement of the temperature control component 2, enhancing the uniformity of temperature transfer along the first direction x, and ensuring uniform heating in each temperature control zone during the relative movement of the amplification section.

[0154] like Figure 7 As shown, in some embodiments, the nucleic acid amplification system further includes: The second detection device is configured to detect the relative positions of the amplification unit and the temperature control component 2 along the first direction x.

[0155] For example, the relative positions include a first relative position, a second relative position, and a third relative position. In the first relative position, the temperature control component 2 is in an open state to allow the microfluidic chip 91 to be inserted. In the second relative position, the temperature control component 2 is in a closed state, with the microfluidic chip 91 located within a second temperature control zone, which is used for the extension phase of the PCR reaction. In the third relative position, the temperature control component 2 is in a closed state, with the microfluidic chip 91 located within a third temperature control zone, which is used for the annealing phase of the PCR reaction.

[0156] like Figure 7 As shown, for example, the second detection device includes multiple photoelectric switches and light baffles 102. The multiple photoelectric switches are disposed on a light-opening plate 103 and arranged at intervals along a first direction x. The light-opening plate 103 is connected to an annular frame 15. The light baffles 102 are connected to a mounting plate 18 and move along the first direction x with the mounting plate 18. During the movement with the mounting plate 18, the light baffles 102 can enter or leave the detection slots of each photoelectric switch to trigger the photoelectric switches to generate position detection signals, which are used to indicate the relative positions of the amplification unit and the temperature control component 2.

[0157] The second detection device can accurately identify and locate the relative positions of the temperature control component 2 and the amplification section along the first direction x, which is beneficial for locating the insertion and removal positions of the microfluidic chip 91, and for accurately positioning the microfluidic chip 91 in each temperature control zone during the PCR reaction, ensuring accurate and reliable reaction position.

[0158] like Figure 7 As shown, in some embodiments, the nucleic acid amplification system further includes: Fiber 8 is configured to emit fluorescence into amplification cavity 90; and The third detection device is used to receive the fluorescence signal within the amplification chamber 90.

[0159] The amplified nucleic acid can be detected by using fiber optic cable 8 and the third detection device, which helps to improve the efficiency of fluorescence signal transmission and detection sensitivity, and ensures that the detection results are accurate and reliable.

[0160] like Figures 7 to 9 As shown, in some embodiments, the temperature control component 2 includes: The light outlet 212 is disposed on the first heat-conducting element 21 and faces the amplification cavity 90. Fluorescence shines into the amplification cavity 90 through the light outlet 212.

[0161] By placing the light outlet 212 on the first heat-conducting component 21, the fluorescence can be directly irradiated into the amplification cavity 90 of the microfluidic chip, shortening the distance between the light-emitting end of the optical fiber and the amplification cavity, reducing the attenuation of the optical signal, thereby improving the sensitivity, accuracy and stability of fluorescence detection.

[0162] In some embodiments, such as Figure 8 As shown, the light outlet 212 is located on one side of the third temperature control zone along the second direction y, and is disposed on the first heat-conducting element 21 used to form the third temperature control zone.

[0163] For PCR reactions, primers only specifically anneal to the target fragment and begin extension during the annealing phase. Positioning the light exit port 212 within the third temperature-controlled zone allows for acquisition under conditions of strong and stable fluorescence signals, helping to avoid interference from high temperatures and ensuring the accuracy and validity of the detection results. Furthermore, this setup enables fluorescence detection when the amplification section is located within the third temperature-controlled zone; that is, while the amplification reagent is in the annealing phase, the PCR reaction and fluorescence detection occur in the same location, thus simplifying the structure and improving spatial integration.

[0164] In addition, the timing and temperature conditions for signal acquisition by the third detection device must be consistent in each PCR cycle. The amplification section stays in the third temperature control zone for a longer time than in the other two temperature control zones. Therefore, setting the light outlet 212 in the third temperature control zone is beneficial for signal acquisition.

[0165] like Figure 10 and Figure 11 As shown, in some embodiments, the first heat-conducting element 21 includes a limiting portion 211, which is located on the side of the first heat-conducting element 21 away from the temperature control zone along the second direction y, and is configured to restrict the movement of the optical fiber 8 toward the temperature control zone along the second direction y.

[0166] For example, the limiting part 211 includes a groove. The groove extends along the second direction y and has an annular sidewall and a bottom wall. The annular sidewall is shaped to fit the optical fiber 8 to restrict the radial movement of the optical fiber 8 within the groove. A light-emitting port 212 is provided on the bottom wall, and the radial dimension of the light-emitting port 212 is smaller than the radial dimension of the bottom wall. After the optical fiber 8 is inserted into the groove, its light-emitting end abuts against the bottom wall, thereby restricting the movement of the optical fiber 8 along the second direction y towards the temperature control zone within the groove.

[0167] The limiting part 211 can restrict the movement of the optical fiber 8 along the second direction y towards the temperature control zone, and prevent the light-emitting end of the optical fiber 8 from moving into the temperature control zone. Therefore, when the temperature control component 2 is in the closed state, the amplification part will not squeeze the optical fiber 8, which is conducive to reliable fluorescence detection.

[0168] like Figure 7 As shown, in some embodiments, the nucleic acid amplification system may include a limiting sheet metal 5 for securing the optical fiber 8 with cable ties or wire clips.

[0169] like Figure 13 and Figure 14 As shown, in some embodiments, the amplification section includes multiple amplification cavities 90, which are arranged in layers, and the light outlet 212 is provided in a one-to-one correspondence with the amplification cavity 90 in the same layer.

[0170] like Figure 13 and Figure 14 As shown, in some embodiments, for example, the amplification section may include four amplification cavities 90, with two amplification cavities 90 distributed in each layer, and the two amplification cavities 90 in each layer arranged along a third direction. The temperature control assembly 2 includes two light-emitting ports 212, which are disposed on the first heat-conducting element 21 for forming the third temperature control zone, for fluorescence collection during the annealing stage.

[0171] Each amplification chamber 90 can accommodate amplification reaction systems corresponding to different viral targets, thereby enabling the parallel detection of multiple viruses.

[0172] like Figures 15 to 17 As shown, in some embodiments, the thermal adjustment unit 23 is located on the first side of the temperature control zone along the second direction y and moves along the first direction x; The main support assembly is in a fixed position along the first direction x and is located on the second side of the temperature control zone along the second direction y. The optical fiber 8 is connected to the main support assembly.

[0173] The thermal conditioning unit 23 is located only on the first side of the amplification section along the second direction y and moves along the first direction x. This allows the optical fiber 8 to be positioned on the second side of the amplification section along the second direction y and fixed in position. This configuration improves the stability of fluorescence emission, thereby enhancing the stability of fluorescence detection and extending the lifespan of the optical fiber 8. Furthermore, the single-sided heating structure reduces the size of the nucleic acid amplification system along the second direction y, allowing for flexible configuration of the nucleic acid amplification system's assembly position.

[0174] like Figures 15 to 18As shown, in some embodiments, the temperature control assembly 2 further includes two temperature control brackets 200, which are arranged opposite to each other and spaced apart along the second direction y. One temperature control bracket 200 is configured to support the heat adjustment part 23 located on the same side of the temperature control zone along the second direction y, and the other temperature control bracket 200 is connected to the main bracket assembly and is configured to support the optical fiber 8. The temperature control bracket 200 connected to the main bracket assembly includes a plurality of first grooves; and / or The amplification section includes a plurality of second grooves 911, which are located on the side of the amplification section facing the main support assembly along the second direction y.

[0175] For example, such as Figure 18 As shown, the microfluidic chip 91 has multiple second grooves 911 arranged in an array.

[0176] The first and second grooves 911 can reduce heat loss from the non-heated side, thereby helping to improve the temperature change rate of the liquid inside the microfluidic chip 91.

[0177] like Figures 15 to 18 As shown, in some embodiments, the main support assembly includes an annular frame 15, to which a temperature control bracket 200 for fixing the optical fiber 8 is connected to the right side frame of the annular frame 15. A heat regulation unit 23 is connected to the temperature control bracket 200 located on the left side, which is connected to a back plate 111. The back plate 111 is erected and connected to the upper rear of the support plate 11. A first drive assembly includes a motor, which is drivenly connected to the support plate 11 and configured to drive the support plate 11 to move along a first direction x, thereby causing the heat regulation unit 23 to move up and down.

[0178] like Figure 15 As shown, in some embodiments, a mounting plate for mounting elastic elements 7 is connected to the left side of the backplate 111, the mounting plate extending in a third direction. The nucleic acid amplification system includes two elastic elements 7, which are spaced apart from each other. The elastic elements 7 are connected between the mounting plate and the temperature control bracket 200 located on the left side, and extend in a second direction y to provide a clamping force to the temperature control bracket 200 in the second direction y.

[0179] like Figure 16 As shown, in some embodiments, the backplate 111 includes a guide rail 13 extending along a first direction x. The main support assembly also includes a guide block 19, which includes a hole that mates with the guide rail 13. The backplate 111 can move along the first direction x through the sliding engagement of the guide rail 13 and the guide block 19.

[0180] like Figure 17 As shown, in some embodiments, the optical fiber 8 can be fixedly connected to the temperature control bracket 200 via the flange 20.

[0181] Another aspect of this disclosure provides a nucleic acid amplification method for a nucleic acid amplification system, wherein the temperature control zone has a target temperature and / or an overshoot temperature, and the nucleic acid amplification method includes: Adjust the temperature of the temperature control zone to the target temperature and / or overshoot temperature; and The amplification unit is controlled to cycle between at least two temperature-controlled zones to achieve nucleic acid amplification.

[0182] The process of pre-adjusting the temperature of at least two temperature control zones includes adjusting the temperature conditions required for each stage of the PCR amplification reaction to the target temperature.

[0183] The first temperature range can be 93℃-98℃, the second temperature range can be 72℃-75℃, and the third temperature range can be 50℃-65℃.

[0184] The nucleic acid amplification method of this disclosure has the advantages of the nucleic acid amplification system of this disclosure. By pre-setting each temperature control zone to the target temperature, it can provide the different temperature conditions required for the denaturation, annealing, and extension stages of the PCR reaction in one go. There is no need to wait for the temperature control zones to rise or fall when switching reaction stages, thereby shortening the cycle time of each PCR reaction and accelerating the amplification process. In addition, this nucleic acid amplification method also helps to reduce temperature fluctuations within the temperature control zone, ensures the stability of heat transfer, and extends the actual effective time of each stage of the PCR reaction, thereby improving nucleic acid amplification efficiency and amplification effect.

[0185] In some embodiments, the target temperature and / or overshoot temperature of the first temperature control zone are in a first temperature range, and the target temperature and / or overshoot temperature of the second temperature control zone are in a second temperature range. Controlling the amplification unit to cyclically switch between at least two temperature control zones includes: The control unit cycles between the first and second temperature control zones.

[0186] This nucleic acid amplification method can reduce PCR reaction time, thereby improving amplification efficiency.

[0187] In some embodiments, the nucleic acid amplification method in one PCR cycle includes: The amplification section is switched to the first temperature control zone and kept in the first temperature control zone until the denaturation stage ends. Set the second temperature control zone to the second overshoot temperature; The control unit is switched to the second temperature control zone; After the temperature difference between the amplification reagent and the second target temperature reaches a preset value, the temperature of the second temperature control zone is adjusted to the second target temperature; and The amplification section is kept in the second temperature control zone until the annealing stage ends.

[0188] In some embodiments, the nucleic acid amplification method in one PCR cycle includes: Set the first temperature control zone to the first overshoot temperature; The amplification unit is switched to the first temperature control zone until the temperature difference between the amplification reagent and the first target temperature reaches the preset difference. Adjust the temperature of the first temperature control zone to the first target temperature, and control the amplification section to remain in the first temperature control zone until the denaturation stage ends; Adjust the temperature of the second temperature control zone to the second overshoot temperature; The amplification unit is switched to the second temperature control zone until the temperature difference between the amplification reagent and the second target temperature reaches a preset value; and Adjust the temperature of the second temperature control zone to the second target temperature, and control the amplification section to remain in the second temperature control zone until the annealing stage ends.

[0189] In some embodiments, the target temperature and / or overshoot temperature of the first temperature control zone is in a first temperature range, the target temperature and / or overshoot temperature of the second temperature control zone is in a second temperature range, and the target temperature and / or overshoot temperature of the third temperature control zone is in a third temperature range. Controlling the amplification unit to cyclically switch between at least two temperature control zones includes: The control amplification unit cycles through the first, second, and third temperature control zones.

[0190] This nucleic acid amplification method can provide different temperature conditions for the denaturation, extension, and annealing stages by using a first, second, and third temperature control zone, respectively. By setting multiple temperature control zones and independently adjusting the temperature of each zone, the temperature fluctuations in each reaction stage can be reduced, making the PCR reaction stable and reliable. This makes the nucleic acid amplification method suitable for clinical nucleic acid testing where high temperature accuracy and reliability are required.

[0191] In some embodiments, pre-adjusting the temperature of at least two temperature control zones includes: Adjust the temperatures of the first, second, and third temperature control zones to the target temperature; Controlling the amplification unit to cycle between at least two temperature-controlled zones includes: The control unit sequentially switches between the first temperature control zone, the third temperature control zone, and the second temperature control zone.

[0192] For example, the temperature of the first temperature control zone is adjusted to the first target temperature, the temperature of the second temperature control zone is adjusted to the second target temperature, and the temperature of the third temperature control zone is adjusted to the third target temperature. The first target temperature is greater than the second and third target temperatures, and the second target temperature is greater than the third target temperature, so that the temperature conditions required for the denaturation stage are formed in the first temperature control zone, the temperature conditions required for the extension stage are formed in the second temperature control zone, and the temperature conditions required for the annealing stage are formed in the third temperature control zone.

[0193] Specifically, the temperature control component 2 and the amplification unit are driven to move relative to each other along the first direction x by the first driving component, so that in each cycle, the amplification unit switches from the first temperature control zone to the third temperature control zone, then to the second temperature control zone, and finally switches back to the first temperature control zone for the next cycle.

[0194] During each cycle switching process, the amplification section is switched sequentially by the first driving component in the order of the first temperature control zone, the third temperature control zone, the second temperature control zone, and finally back to the first temperature control zone. This ensures that the PCR reaction proceeds stably and strictly in the order of denaturation, annealing, and extension, guaranteeing the continuous and reliable PCR reaction process. It also helps to avoid disordered reaction order, thereby improving the stability of nucleic acid amplification and the reproducibility of amplification results.

[0195] In some embodiments, pre-adjusting the temperature of at least two temperature control zones includes: Adjust the temperatures of the first and third temperature control zones to the target temperature; and Set the second temperature control zone to overshoot temperature.

[0196] Controlling the amplification unit to cycle between at least two temperature-controlled zones includes: During each cycle switching process, the control amplification unit switches sequentially from the first temperature control zone to the second temperature control zone, then to the third temperature control zone, and finally back to the first temperature control zone.

[0197] The overshoot temperature of the second temperature control zone is lower than the target temperature of the third temperature control zone.

[0198] This nucleic acid amplification method utilizes a second temperature control zone to set an overshoot temperature for the annealing stage in each cycle. This accelerates the cooling rate of the amplification reagents before entering the annealing stage, which helps ensure the temperature stability of the amplification reagents within the third temperature control zone, reduces temperature fluctuations, and extends the time the amplification section spends in the annealing stage. This, in turn, facilitates a stable and reliable annealing process.

[0199] In some embodiments, both the second and third temperature control zones may be set with an overshoot temperature lower than the target temperature of the third temperature control zone to prolong the time the amplification section is in a low-temperature environment, thereby improving the stability of the annealing reaction. For example, the target temperature of the third temperature control zone is 55°C, and the overshoot temperature of the second and third temperature control zones is 50°C.

[0200] In some embodiments, controlling the amplification unit to cycle between at least two temperature control zones includes: During each cycle switching process, the control amplification unit switches sequentially from the first temperature control zone to the third temperature control zone, then to the second temperature control zone, and finally back to the first temperature control zone. Pre-setting the temperature of at least two temperature control zones includes: If the temperature control zone only has the target temperature, adjust the temperature of the temperature control zone to the target temperature; and Within the temperature control zone, which has both a target temperature and an overshoot temperature, the temperature of the temperature control zone is first adjusted to the overshoot temperature, and then adjusted to the target temperature.

[0201] Specifically, the overshoot temperature of the first temperature control zone can be greater than the target temperature of the first temperature control zone, the overshoot temperature of the second temperature control zone can be greater than the target temperature of the second temperature control zone, and the overshoot temperature of the third temperature control zone is lower than the target temperature of the third temperature control zone. The temperature difference between the overshoot temperature and the target temperature of each temperature control zone is, for example, 1℃-5℃.

[0202] This nucleic acid amplification method accelerates the rate at which the amplification reagents reach the target temperature in each temperature-controlled zone by setting an overshoot temperature for each zone, thereby improving the amplification efficiency of the PCR reaction. Furthermore, this method extends the time the amplification reagents spend in a low-temperature environment, which further prolongs the time the amplification section is in the annealing phase, thus ensuring a stable and reliable annealing process.

[0203] Another aspect of this disclosure provides a nucleic acid detection method using a nucleic acid amplification system, comprising: Emit fluorescence into amplification chamber 90; and Detect the fluorescence signal within the amplification chamber 90.

[0204] This nucleic acid detection method enables real-time and accurate detection of nucleic acid amplification results by emitting fluorescence into the amplification chamber 90 and detecting the fluorescence signal, which helps to ensure the accuracy and reliability of the detection results.

[0205] In some embodiments, the amplification unit includes a plurality of amplification cavities 90, which are arranged in layers, and fluorescence is emitted into the amplification cavities 90 layer by layer and the fluorescence signal in the amplification cavities 90 is detected.

[0206] This nucleic acid detection method can detect multiple viruses in parallel, which helps to improve the efficiency of nucleic acid detection.

[0207] by Figure 13 and Figure 14 Taking the nucleic acid amplification system shown as an example, the amplification section may include four amplification chambers 90, of which two amplification chambers 90 are located at the top layer and the other two amplification chambers 90 are located at the bottom layer.

[0208] The temperature control component 2 is driven to move to the first detection position, so that the two light outlets 212 are aligned with the two amplification cavities 90 located on the top layer, and fluorescence is collected; the temperature control component 2 is driven to move down to the second detection position, so that the two light outlets 212 are aligned with the two amplification cavities 90 located on the bottom layer, and fluorescence is collected again.

[0209] Another aspect of this disclosure provides a nucleic acid detection system, which includes the nucleic acid amplification system provided in this disclosure.

[0210] The nucleic acid detection system of this disclosure has the advantages of the nucleic acid amplification system of this disclosure.

[0211] The following example, using a nucleic acid amplification system with three temperature-controlled zones, illustrates the nucleic acid amplification method in detail: (1) Set three temperature control zones with only target temperature: Set the first temperature control zone to the first target temperature, the second temperature control zone to the second target temperature, and the third temperature control zone to the third target temperature. The amplification section is switched to the first temperature control zone and kept in the first temperature control zone until the denaturation stage ends. The amplification section is switched to the third temperature control zone and kept in the third temperature control zone until the annealing stage ends. The amplification section is switched to the second temperature control zone and kept in the second temperature control zone until the extension stage ends. The control unit switches back to the first temperature control zone to begin the next cycle.

[0212] (2) Set the first and third temperature control zones to have target temperatures, and the second temperature control zone to have an overshoot temperature, with the overshoot temperature of the second temperature control zone being lower than the target temperature of the third temperature control zone (suitable for amplification of short DNA fragments): Adjust the temperatures of the first and third temperature control zones to their respective target temperatures, and adjust the temperature of the second temperature control zone to the overshoot temperature; The amplification section is switched to the first temperature control zone and kept in the first temperature control zone until the denaturation stage ends. The amplification unit is switched to the second temperature control zone until the temperature difference between the amplification reagent and the third target temperature reaches the preset difference. The amplification section is switched to the third temperature control zone and kept in the third temperature control zone until the annealing stage ends. The control unit switches back to the first temperature control zone to begin the next cycle.

[0213] (3) A second temperature control zone (as a temperature control zone for extending the reaction) is set up, which has both the target temperature and the overshoot temperature: Before the amplification section switches from the first temperature control zone to the second temperature control zone, the second temperature control zone is adjusted to the overshoot temperature; When the temperature difference between the amplification section and the target temperature of the second temperature control zone reaches the preset difference, the second temperature control zone is then adjusted to the target temperature.

[0214] The adjustment methods for the first and third temperature control zones, which simultaneously have target temperature and overshoot temperature, are the same as those for the second temperature control zone, and will not be repeated here.

[0215] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of this disclosure and not to limit them; although this disclosure has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications can still be made to the specific implementation of this disclosure or equivalent substitutions can be made to some technical features, all of which should be covered within the scope of the technical solutions claimed in this disclosure.

Claims

1. A nucleic acid amplification system, characterized in that, include: The microfluidic cartridge (9) includes an amplification section having an amplification cavity (90) for containing amplification reagents. At least two temperature control zones are arranged along a first direction (x), the temperature control zones are configured to accommodate the amplification unit, and the temperature of at least a portion of the temperature control zones is adjustable; The temperature control component (2) is configured to independently adjust the temperature of each of the temperature control zones so that the temperatures of each of the temperature control zones are different; and A first driving component, drivenly connected to the amplification unit and / or the temperature control component (2), is configured to move the amplification unit and the temperature control component (2) relative to each other along the first direction (x) so that the amplification chamber (90) cycles between at least two of the temperature control zones.

2. The nucleic acid amplification system according to claim 1, characterized in that, The at least two temperature control zones include a first temperature control zone and a second temperature control zone. The temperature of the first temperature control zone is in a first temperature range, and the temperature of the second temperature control zone is in a second temperature range. The temperature value of the first temperature range is generally higher than the temperature value of the second temperature range.

3. The nucleic acid amplification system according to claim 1, characterized in that, The at least two temperature control zones include a first temperature control zone, a second temperature control zone, and a third temperature control zone. The temperature of the first temperature control zone is in a first temperature range, the temperature of the second temperature control zone is in a second temperature range, and the temperature of the third temperature control zone is in a third temperature range. The temperature value of the first temperature range is higher than the temperature values ​​of the second and third temperature ranges, and the second temperature range has a segment that is higher than the third temperature range.

4. The nucleic acid amplification system according to any one of claims 1-3, characterized in that, The temperature control component (2) includes a heat adjustment section (23) located on at least one side of the temperature control zone along a second direction (y) and configured to heat or cool the temperature control zone; wherein the second direction (y) is perpendicular to the first direction (x).

5. The nucleic acid amplification system according to claim 4, characterized in that, The temperature control component (2) includes multiple sets of heat adjustment parts (23) arranged along a first direction (x), each set of heat adjustment parts (23) includes two heat adjustment parts (23) arranged opposite to each other and spaced apart along a second direction (y), and a temperature control zone is formed between the two heat adjustment parts (23).

6. The nucleic acid amplification system according to claim 5, characterized in that, Also includes: The second drive assembly, which is driven connected to multiple sets of the thermal adjustment units (23), is configured to drive the thermal adjustment units (23) on both sides of the temperature control zone to move along the second direction (y) to move closer to each other.

7. The nucleic acid amplification system according to claim 6, characterized in that, The temperature control assembly (2) further includes two temperature control brackets (200), which are arranged opposite to each other and spaced apart along the second direction (y). The temperature control brackets (200) are configured to support the heat adjustment part (23) located on the same side of the temperature control zone along the second direction (y). The second drive assembly includes a plurality of elastic elements (7), and at least one elastic element (7) is connected to the side of each temperature control bracket (200) away from the temperature control zone along the second direction (y). The elastic element (7) extends along the second direction (y) and is configured to apply pressure along the second direction (y) toward the temperature control zone to the connected temperature control bracket (200).

8. The nucleic acid amplification system according to claim 6, characterized in that, Also includes: The first guide mechanism is configured to guide the movement of the thermal adjustment section (23) on both sides of the temperature control zone along the second direction (y).

9. The nucleic acid amplification system according to claim 5, characterized in that, The temperature control assembly (2) further includes two temperature control brackets (200), which are arranged opposite to each other and spaced apart along the second direction (y). The temperature control brackets (200) are configured to support the heat adjustment part (23) located on the same side of the temperature control zone along the second direction (y). The temperature control brackets (200) include a first mating part (16). The nucleic acid amplification system also includes a main support assembly, the main support assembly including a second mating part (101), the first driving assembly being configured to drive the temperature control support (200) to move relative to the main support assembly along the first direction (x) so that the first mating part (16) moves along the first direction (x) closer to the second mating part (101), the second mating part (101) being configured to apply pressure to the first mating part (16) along the second direction (y) away from the temperature control zone.

10. The nucleic acid amplification system according to claim 4, characterized in that, The temperature control component (2) also includes: A plurality of first heat-conducting elements (21), corresponding to a plurality of said heat-regulating sections (23), are located on the side of said heat-regulating section (23) along the second direction (y) close to said temperature control zone, and are configured to transfer the cold or heat released by said heat-regulating section (23) to said temperature control zone; and / or Multiple second heat-conducting elements are disposed corresponding to multiple heat-regulating parts (23) and multiple first heat-conducting elements (21), and the second heat-conducting elements are connected along the second direction (y) between the corresponding first heat-conducting element (21) and the heat-regulating part (23) to form a continuous heat transfer path.

11. The nucleic acid amplification system according to claim 4, characterized in that, The temperature control component (2) also includes: Multiple heat insulation elements (24) are provided corresponding to multiple heat regulation parts (23), and the heat insulation elements (24) are located on the side of the corresponding heat regulation part (23) away from the temperature control zone along the second direction (y).

12. The nucleic acid amplification system according to claim 4, characterized in that, The temperature control component (2) also includes: The first detection device (27) is configured to detect the temperature value of the thermal regulation unit (23); and The control device is signal-connected to the first detection device (27) and the thermal regulation unit (23), and is configured to control the working state of the thermal regulation unit (23) according to the detection result of the first detection device (27).

13. The nucleic acid amplification system according to claim 12, characterized in that, The temperature control component (2) also includes: The first circuit board (201) is electrically connected to the thermal regulation unit (23) and the first detection device (27), and is signal-connected to the control device. It is configured to transmit the detection signal of the first detection device (27) to the control device and the control signal of the control device to the thermal regulation unit (23).

14. The nucleic acid amplification system according to claim 13, characterized in that, The temperature control component (2) also includes: The second circuit board (203) is perpendicular to the first circuit board (201). The second circuit board (203) is electrically connected to the first circuit board (201) via pin headers and is also signal-connected to the control device.

15. The nucleic acid amplification system according to any one of claims 1-3, characterized in that, Also includes: The heat dissipation device (3) is configured to dissipate heat from the temperature control zone when the temperature in the temperature control zone exceeds a preset threshold.

16. The nucleic acid amplification system according to any one of claims 1-3, characterized in that, Also includes: The second guiding mechanism is configured to guide the relative movement of the amplification section and the temperature control component (2) along the first direction (x).

17. The nucleic acid amplification system according to any one of claims 1-3, characterized in that, Also includes: The second detection device is configured to detect the relative position of the amplification unit and the temperature control component (2) along the first direction (x).

18. The nucleic acid amplification system according to any one of claims 1-3, characterized in that, Also includes: An optical fiber (8) is configured to emit fluorescence into the amplification cavity (90); and The third detection device is used to receive the fluorescence signal in the amplification cavity (90).

19. The nucleic acid amplification system according to claim 18, characterized in that, The temperature control component (2) includes a heat adjustment unit (23), which is located on a first side of the temperature control zone along a second direction (y) and moves along the first direction (x). The heat adjustment unit (23) is configured to heat or cool the temperature control zone. The nucleic acid amplification system further includes a main support assembly, which is located at a fixed position along the first direction (x) and on the second side of the temperature control zone along the second direction (y), and the optical fiber (8) is connected to the main support assembly; Wherein, the second direction (y) is perpendicular to the first direction (x).

20. The nucleic acid amplification system according to claim 18, characterized in that, The temperature control assembly (2) further includes two temperature control brackets (200), which are arranged opposite to each other and spaced apart along the second direction (y). One of the temperature control brackets (200) is configured to support the heat adjustment part (23) located on the same side of the temperature control zone along the second direction (y), and the other temperature control bracket (200) is connected to the main bracket assembly and is configured to support the optical fiber (8). The temperature control bracket (200) connected to the main support assembly includes a plurality of first grooves; and / or The amplification section includes a plurality of second grooves (911), the second grooves (911) being located on one side of the amplification section facing the main support assembly along the second direction (y).

21. The nucleic acid amplification system according to claim 18, characterized in that, The temperature control component (2) includes: The thermal regulation unit (23) is configured to heat or cool the temperature control zone; and The first heat-conducting element (21) is provided corresponding to the heat regulation part (23) and is located on the side of the heat regulation part (23) close to the temperature control zone along the second direction (y), and the second direction (y) is perpendicular to the first direction (x). A light-emitting port (212) is disposed on the first heat-conducting element (21) and faces the amplification cavity (90), and the fluorescence shines onto the amplification cavity (90) through the light-emitting port (212).

22. The nucleic acid amplification system according to claim 21, characterized in that, The first heat-conducting element (21) includes a limiting part (211) located on the side of the first heat-conducting element (21) away from the temperature control zone along the second direction (y), and is configured to restrict the optical fiber (8) from moving closer to the temperature control zone along the second direction (y).

23. The nucleic acid amplification system according to claim 21, characterized in that, The amplification section includes a plurality of amplification cavities (90), which are arranged in layers, and the light outlet (212) is provided in a one-to-one correspondence with the amplification cavity (90) in the same layer.

24. A nucleic acid detection system, characterized in that, The nucleic acid amplification system includes any one of claims 1 to 23.

25. A nucleic acid amplification method based on the nucleic acid amplification system according to any one of claims 1 to 23, characterized in that, The temperature control zone has a target temperature and / or an overshoot temperature, and the nucleic acid amplification method includes: Adjust the temperature of the temperature control zone to the target temperature and / or overshoot temperature; and The amplification unit is controlled to cyclically switch between at least two of the temperature-controlled zones to achieve nucleic acid amplification.

26. The nucleic acid amplification method according to claim 25, characterized in that, The at least two temperature control zones include a first temperature control zone and a second temperature control zone. The target temperature and / or the overshoot temperature of the first temperature control zone is within a first temperature range, and the target temperature and / or the overshoot temperature of the second temperature control zone is within a second temperature range. The temperature value of the first temperature range is higher than the temperature value of the second temperature range. Controlling the amplification unit to cyclically switch between the at least two temperature control zones includes: The amplification unit is controlled to cycle between the first temperature control zone and the second temperature control zone.

27. The nucleic acid amplification method according to claim 25, characterized in that, The at least two temperature control zones include a first temperature control zone, a second temperature control zone, and a third temperature control zone. The target temperature and / or the overshoot temperature of the first temperature control zone is within a first temperature range. The target temperature and / or the overshoot temperature of the second temperature control zone is within a second temperature range. The target temperature and / or the overshoot temperature of the third temperature control zone is within a third temperature range. The temperature value of the first temperature range is higher than the temperature values ​​of the second and third temperature ranges. The second temperature range has a segment that is higher than the third temperature range. Controlling the amplification unit to cyclically switch between the at least two temperature control zones includes: The amplification unit is controlled to cycle between the first temperature control zone, the second temperature control zone, and the third temperature control zone.

28. The nucleic acid amplification method according to claim 27, characterized in that, Pre-adjusting the temperature of at least two of the temperature control zones includes: Adjust the temperatures of the first temperature control zone, the second temperature control zone, and the third temperature control zone to the target temperature; Controlling the amplification unit to cycle between at least two of the temperature control zones includes: The amplification unit is controlled to sequentially switch between the first temperature control zone, the third temperature control zone, and the second temperature control zone.

29. The nucleic acid amplification method according to claim 27, characterized in that, Pre-adjusting the temperature of at least two of the temperature control zones includes: Adjust the temperatures of the first temperature control zone and the third temperature control zone to the target temperature; and Adjust the second temperature control zone to the overshoot temperature; Controlling the amplification unit to cycle between at least two of the temperature control zones includes: During each cycle switching process, the amplification unit is controlled to switch sequentially from the first temperature control zone to the second temperature control zone, then to the third temperature control zone, and finally back to the first temperature control zone.

30. The nucleic acid amplification method according to claim 27, characterized in that, Controlling the amplification unit to cycle between at least two of the temperature control zones includes: During each cycle switching process, the amplification unit is controlled to switch sequentially from the first temperature control zone to the third temperature control zone, then to the second temperature control zone, and finally back to the first temperature control zone; Pre-adjusting the temperature of at least two of the temperature control zones includes: When the temperature control zone is only at the target temperature, adjust the temperature of the temperature control zone to the target temperature; and When the temperature control zone has the target temperature and the overshoot temperature, the temperature of the temperature control zone is first adjusted to the overshoot temperature, and then adjusted to the target temperature.

31. A nucleic acid detection method based on the nucleic acid amplification system according to any one of claims 1 to 23, characterized in that, include: Fluorescence is emitted into the amplification chamber (90); and The fluorescence signal within the amplification chamber (90) is detected.

32. The nucleic acid detection method according to claim 31, characterized in that, The amplification section includes a plurality of amplification cavities (90), which are arranged in layers. Fluorescence is emitted into the amplification cavities (90) layer by layer and the fluorescence signal in the amplification cavities (90) is detected.

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