An automated, non-amplification nucleic acid detection device

Abstract

The application belongs to the technical field of biomedical engineering, and relates to an automatic non-amplification nucleic acid detection device, which comprises a mounting assembly, a first fluid input assembly, a second fluid input assembly, a third fluid input assembly, a fourth fluid input assembly, a detection assembly, a fluid output assembly and a control assembly. The automatic non-amplification nucleic acid detection device provided in the application has excellent sensitivity, good specificity and wide applicability, so that high-sensitivity automatic nucleic acid molecule detection is possible.

Description

Technical Field

[0001] This invention relates to the field of biomedical engineering technology, and in particular to an automated amplification-free nucleic acid detection device. Background Technology

[0002] Emerging molecular diagnostics is fundamentally transforming healthcare, providing more precise analysis for disease diagnosis, prognosis, and management. Among numerous molecular diagnostic tools, the CRISPR (a repetitive sequence within the prokaryotic genome) / Cas enzyme (a cysteine-containing aspartate-specific protease) system, which uses programmable RNA-guided (gRNA) DNA endonucleases to target specific sequences, demonstrates great potential as a novel detection platform. However, despite its groundbreaking prospects, this technology still has limitations in sensitivity, especially in detecting low-abundance targets, which is crucial for the early diagnosis and monitoring of diseases.

[0003] Combining CRISPR / Cas12a with nucleic acid amplification technologies, such as polymerase chain reaction (PCR), recombinase polymerase amplification (RPA), or loop-mediated isothermal amplification (LAMP), has become a routine method for improving diagnostic sensitivity. However, this demand highlights a key bottleneck in molecular diagnostics—the inherent limitations of amplification techniques. The amplification process is prone to errors, including nonspecific amplification and contamination risks, thus affecting diagnostic accuracy. Furthermore, the complex thermal cycling equipment required for PCR or the precise temperature control of LAMP increases operational complexity and cost, making these methods unsuitable for point-of-care testing (POCT) in resource-constrained or field-based environments. Therefore, while amplification techniques have helped improve the sensitivity of CRISPR technology, they have also introduced new limitations that hinder the widespread application and adoption of molecular diagnostics.

[0004] Ideal molecular diagnostic point-of-care testing (POCT) devices should be automated, user-friendly, and cost-effective. These devices should not only be able to handle the complexities of genetic and molecular analysis but also integrate seamlessly with the emerging digital health ecosystem. Automation and simplification of diagnostic procedures are crucial to ensuring the availability and widespread use of these tools in resource-limited environments and non-traditional healthcare facilities. However, current amplification-based diagnostic methods fail to meet these requirements due to their operational complexity, high cost, and the need for specialized equipment and training.

[0005] The CRISPR / Cas12a system presents challenges in achieving ultrasensitive automated detection of nucleic acids. To build a nucleic acid detection device with ultrasensitive, automated, and rapid detection capabilities, the following issues need to be addressed:

[0006] i) The CRISPR / Cas12a system has limitations in the sensitivity of nucleic acid detection, especially when detecting low-abundance targets;

[0007] ii) The integration process of CRISPR / Cas12a with nucleic acid amplification technology is complex and makes it difficult to achieve real-time nucleic acid detection;

[0008] iii) The automation of CRISPR / Cas12a for nucleic acid diagnostics has not yet been fully realized. Summary of the Invention

[0009] In view of this, embodiments of this application provide an automated amplification-free nucleic acid detection device.

[0010] Specifically, the present invention is achieved through the following technical solution:

[0011] According to a first aspect of the present invention, an automated amplification-free nucleic acid detection device is provided, comprising:

[0012] Installation components are used to provide installation space;

[0013] A first fluid input component is used to input a sample solution; the first fluid input component is disposed on the mounting component;

[0014] A second fluid input component is used to input the detection reagent solution; the second fluid input component is disposed on the mounting component.

[0015] A third fluid input component is used to input a flushing solution; the third fluid input component is disposed on the mounting component;

[0016] A fourth fluid input component is used to input a protein streptavidin solution; the fourth fluid input component is disposed on the mounting component.

[0017] A detection component is used to detect the presence of nucleic acids in a sample solution; the detection component is disposed on the mounting component and is connected to the first fluid input component, the second fluid input component, the third fluid input component, and the fourth fluid input component, respectively;

[0018] A fluid output component for outputting the post-reaction mixed solution; the fluid output component is disposed on the mounting component and connected to the detection component;

[0019] A control component is used to control the various parts; the control component is disposed on the mounting component and is connected to the first fluid input component, the second fluid input component, the third fluid input component, the fourth fluid input component, the detection component, and the fluid output component, respectively.

[0020] Optionally, the mounting assembly includes a housing and a mounting plate, wherein the mounting plate is disposed inside the housing, and the first fluid input assembly, the second fluid input assembly, the third fluid input assembly, the fourth fluid input assembly, the detection assembly, the fluid output assembly, and the control assembly are respectively disposed on the mounting plate.

[0021] Optionally, the first fluid input component includes a first micropump and a first fluid delivery pipe, wherein the first micropump is connected to the control component and the first fluid delivery pipe, and the first fluid delivery pipe is connected to the detection component.

[0022] Optionally, the second fluid input component includes: a second micropump and a second fluid delivery pipe, wherein the second micropump is connected to the control component and the second fluid delivery pipe respectively, and the second fluid delivery pipe is connected to the detection component.

[0023] Optionally, the third fluid input component includes a third micropump and a third fluid delivery pipe, wherein the third micropump is connected to the control component and the third fluid delivery pipe respectively, and the third fluid delivery pipe is connected to the detection component.

[0024] Optionally, the fourth fluid input component includes a fourth micropump and a fourth fluid delivery pipe, wherein the fourth micropump is connected to the control component and the fourth fluid delivery pipe respectively, and the fourth fluid delivery pipe is connected to the detection component.

[0025] Optionally, the detection component includes: a first reaction chamber, a second reaction chamber, and a third reaction chamber, wherein the first reaction chamber is connected to a first fluid delivery pipe in the first fluid input component, a second fluid delivery pipe in the second fluid input component, a fourth fluid delivery pipe in the fourth fluid input component, and the second reaction chamber, respectively; the third reaction chamber is connected to a third fluid delivery pipe in the third fluid input component, the fluid output component, and the second reaction chamber, respectively; a heater is provided on the bottom wall of the first reaction chamber, and the heater is connected to the control component; an electrode is provided on the bottom wall of the third reaction chamber, and the electrode is connected to the control component.

[0026] Optionally, the fluid output assembly includes a fluid output pipe, wherein the fluid output pipe is connected to a third reaction chamber in the detection assembly.

[0027] Optionally, the control component includes: a microfluidic control module, a heating controller, a micropump controller, and a wireless transmission module, wherein the microfluidic control module is connected to the heating controller and the micropump controller, the heating controller is connected to the heater in the detection component, the micropump controller is connected to the first micropump in the first fluid input component, the second micropump in the second fluid input component, the third micropump in the third fluid input component, and the fourth micropump in the fourth fluid input component, and the wireless transmission module is connected to the electrodes in the detection component.

[0028] Optionally, the control component further includes a transmission end, which is connected to the wireless transmission module and the electrode respectively.

[0029] The technical solution provided in this application has at least the following beneficial effects:

[0030] This application provides an automated amplification-free nucleic acid detection device, including an installation component that provides the necessary installation space, supports, and integrates all other components. A first fluid input component is used to input a sample solution containing the nucleic acid to be detected. A second fluid input component is used to input a detection reagent solution, which may react with a specific nucleic acid sequence in the sample solution to aid detection. A third fluid input component is used to input a rinsing solution for cleaning the system and ensuring the accuracy of the detection results. A fourth fluid input component is used to input a protein streptavidin solution, which may be used to bind to specifically labeled nucleic acids to enhance the detection signal. A detection component is the core component used to detect the nucleic acid status in the sample solution. It receives and processes fluids by connecting to all fluid input components to perform nucleic acid detection. A fluid output component is used to output the mixed solution after the reaction, which may be waste liquid or a solution requiring further analysis. A control component is responsible for controlling the operation of all other components, ensuring the automation and accuracy of the entire detection process.

[0031] By precisely controlling the fluid input and detection process, the device can detect very low concentrations of nucleic acids, improving detection sensitivity. Using specific detection reagents and a protein streptavidin solution, the device can selectively detect target nucleic acid sequences, reducing false alarms. The device's design allows it to be applied to a variety of nucleic acid detection scenarios, suitable for different types of samples and nucleic acid sequences. The integration of control components automates the entire detection process, reducing the need for manual operation and improving efficiency and accuracy. This automated, amplification-free nucleic acid detection device achieves a fully automated process from sample input to result output by integrating multiple functional components. Its high sensitivity, specificity, and broad applicability make it an ideal tool for high-precision nucleic acid detection. The device's design not only improves detection efficiency but also reduces operational complexity, enabling the automation and widespread adoption of nucleic acid molecular detection. Attached Figure Description

[0032] The accompanying drawings, which are incorporated in and form part of this specification, illustrate embodiments consistent with the invention and, together with the description, serve to explain the principles of the invention.

[0033] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the description of the embodiments or related technologies will be briefly introduced below. Obviously, those skilled in the art can obtain other drawings based on these drawings without creative effort.

[0034] Figure 1 This is a schematic diagram of the structure of an automated amplification-free nucleic acid detection device provided in an embodiment of this application;

[0035] Figure 2 This is a schematic diagram of the structure of an automated amplification-free nucleic acid detection device provided in an embodiment of this application;

[0036] Figure 3 This is a partial structural schematic diagram of an automated amplification-free nucleic acid detection device provided in an embodiment of this application;

[0037] Figure 4 A schematic diagram of the structure of the detection component in an automated amplification-free nucleic acid detection device provided in this application embodiment. Detailed Implementation

[0038] To make the objectives, technical solutions, and advantages of the embodiments of this application clearer, the technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0039] Figure 1 An automated amplification-free nucleic acid detection device applicable to embodiments of this application is illustrated schematically.

[0040] Reference Figure 1-4 As shown, an automated amplification-free nucleic acid detection device is provided, comprising:

[0041] Install component 10, which provides installation space;

[0042] A first fluid input component 20 is used to input a sample solution; the first fluid input component 20 is disposed on the mounting component 10;

[0043] The second fluid input component 30 is used to input the detection reagent solution; the second fluid input component 30 is disposed on the mounting component 10.

[0044] A third fluid input component 40 is used to input a flushing solution; the third fluid input component 40 is disposed on the mounting component 10;

[0045] A fourth fluid input component 50 is used to input a protein streptavidin solution; the fourth fluid input component 50 is disposed on the mounting component 10.

[0046] The detection component 60 is used to detect the nucleic acid status in the sample solution; the detection component 60 is disposed on the mounting component 10 and is connected to the first fluid input component 20, the second fluid input component 30, the third fluid input component 40 and the fourth fluid input component 50 respectively;

[0047] A fluid output component 70 is used to output the mixed solution after the reaction; the fluid output component 70 is disposed on the mounting component 10 and connected to the detection component 60;

[0048] A control component 80 is used to control each component; the control component 80 is disposed on the mounting component 10 and is respectively connected to the first fluid input component 20, the second fluid input component 30, the third fluid input component 40, the fourth fluid input component 50, the detection component 60 and the fluid output component 70.

[0049] In this embodiment, a sample solution containing the sample to be tested is input into the detection component 60 via a first fluid input component 20. A detection reagent solution (containing Cas12a, gRNA, and single-stranded reporter DNA modified with fluorophore (FAM) and biotinylate molecule (ssDNA-FB)) is input into the detection component 60 via a second fluid input component 30. A streptavidin (SA) solution is input into the detection component 60 via a fourth fluid input component 50. A PBS buffer (dissolving protective reagent) is input into the detection component 60 via the first fluid input component 20. The solutions react in the detection component 60. The detection component 60 detects the nucleic acid status in the sample solution and outputs the detection results. Then, the fluid outputs the mixed solution after reaction via a fluid output component 70. The control component 80 controls the normal operation of each component in the device.

[0050] For example, the mounting assembly 10 includes a housing 11 and a mounting plate 12, wherein the mounting plate 12 is disposed inside the housing 11, and the first fluid input assembly 20, the second fluid input assembly 30, the third fluid input assembly 40, the fourth fluid input assembly 50, the detection assembly 60, the fluid output assembly 70 and the control assembly 80 are respectively disposed on the mounting plate 12.

[0051] In this embodiment, the housing 11 is used to house and protect the various parts, and the mounting plate 12 is used to mount the various parts.

[0052] For example, the first fluid input component 20 includes a first micro pump 21 and a first fluid delivery pipe 22, wherein the first micro pump 21 is connected to the control component 80 and the first fluid delivery pipe 22 respectively, and the first fluid delivery pipe 22 is connected to the detection component 60.

[0053] In this embodiment, the first fluid delivery tube 22 is connected to the sample solution, and the first micro pump 21 receives the control command from the control component 80 and drives the sample solution into the detection component 60.

[0054] For example, the second fluid input component 30 includes a second micro pump 31 and a second fluid delivery pipe 32, wherein the second micro pump 31 is connected to the control component 80 and the second fluid delivery pipe 32 respectively, and the second fluid delivery pipe 32 is connected to the detection component 60.

[0055] In this embodiment, the second fluid delivery tube 32 is connected to the detection reagent solution (containing Cas12a, gRNA, and single-stranded reporter DNA modified with fluorophore (FAM) and biotin molecules (ssDNA-FB), and the second micropump 31 receives control commands from the control component 80 and drives the sample solution into the detection component 60.

[0056] For example, the third fluid input component 40 includes a third micro pump 41 and a third fluid delivery pipe 42, wherein the third micro pump 41 is connected to the control component 80 and the third fluid delivery pipe 42 respectively, and the third fluid delivery pipe 42 is connected to the detection component 60.

[0057] In this embodiment, the third fluid delivery tube 42 is connected to the PBS buffer, and the third micropump 41 receives the control command from the control component 80 and drives the sample solution into the detection component 60.

[0058] For example, the fourth fluid input component 50 includes a fourth micro pump 51 and a fourth fluid delivery pipe 52, wherein the fourth micro pump 51 is connected to the control component 80 and the fourth fluid delivery pipe 52 respectively, and the fourth fluid delivery pipe 52 is connected to the detection component 60.

[0059] In this embodiment, the fourth fluid delivery tube 52 is connected to the streptavidin (SA) solution, and the fourth micropump 51 receives the control command from the control component 80 and drives the sample solution into the detection component 60.

[0060] For example, the detection component 60 includes a first reaction chamber 61, a second reaction chamber 62, and a third reaction chamber 63. The first reaction chamber 61 is connected to the first fluid delivery pipe 22 in the first fluid input component 20, the second fluid delivery pipe 32 in the second fluid input component 30, the fourth fluid delivery pipe 52 in the fourth fluid input component 50, and the second reaction chamber 62. The third reaction chamber 63 is connected to the third fluid delivery pipe 42 in the third fluid input component 40, the fluid output component 70, and the second reaction chamber 62. A heater is provided on the bottom wall of the first reaction chamber 61, and the heater is connected to the control component 80. An electrode is provided on the bottom wall of the third reaction chamber 63, and the electrode is connected to the control component 80.

[0061] In this embodiment, the first reaction chamber 61, the second reaction chamber 62, and the third reaction chamber 63 are connected by a serpentine channel. A first micropump 21 drives the sample solution into the serpentine channel, and a second micropump 31 drives the detection reagent solution (containing Cas12a, gRNA, and single-stranded reporter DNA modified with a fluorophore (FAM) and biotin molecules (ssDNA-FB)) into the serpentine channel, then into the first reaction chamber 61. The reaction is carried out at 37°C for 15 minutes, followed by a reaction at 65°C for 5 minutes. The first and second micropumps 21 and 31 simultaneously drive the reagent in the first reaction chamber 61, while a fourth micropump 51 drives the streptavidin (SA) solution through the serpentine channel into the second reaction chamber 62 at room temperature for 15 minutes. The first, second, and fourth micropumps 21, 31, and 51 drive the liquid in the second reaction chamber 62 to contact the electrode surface for 20 minutes. A third micropump 41 drives PBS buffer to rinse the electrode in the third reaction chamber 63. A heater, controlled by a control component 80, heats the solution.

[0062] For example, the fluid output assembly 70 includes a fluid output pipe 71, wherein the fluid output pipe 71 is connected to the third reaction chamber 63 in the detection assembly 60.

[0063] In this embodiment of the application, the mixed solution is discharged via fluid output pipe 71.

[0064] For example, the control component 80 includes: a microfluidic control module 81, a heating controller 82, a micropump controller 83, and a wireless transmission module 84. The microfluidic control module 81 is connected to the heating controller 82 and the micropump controller 83, respectively. The heating controller 82 is connected to the heater in the detection component 60. The micropump controller 83 is connected to the first micropump 21 in the first fluid input component 20, the second micropump 31 in the second fluid input component 30, the third micropump 41 in the third fluid input component 40, and the fourth micropump 51 in the fourth fluid input component 50, respectively. The wireless transmission module 84 is connected to the electrodes in the detection component 60.

[0065] In this embodiment, the microfluidic control module 81 stores control commands, which are used to send control commands to the heating controller 82 and the micropump controller 83 respectively. The heating controller 82 controls the heater on the bottom wall of the first reaction chamber 61 to heat it. The micropump controller 83 controls the opening or closing of the first micropump 21, the second micropump 31, the third micropump 41 and the fourth micropump 51 respectively. The detection signal of the electrode is sent to the terminal through the wireless transmission module 84 for the testing personnel to read.

[0066] For example, the control component 80 further includes a transmission terminal 85, which is connected to the wireless transmission module 84 and the electrode respectively.

[0067] In this embodiment, the detection signal from the electrode is transmitted to the wireless transmission module 84 via the transmission terminal 85, and then sent to the terminal for the testing personnel to read. If nucleic acid is present in the sample solution, the detection signal from the electrode will change. Similarly, the nucleic acid content in the sample will also affect the detection signal from the electrode. Therefore, the detection signal from the electrode will indicate the presence or absence of nucleic acid in the sample solution and the content of nucleic acid, and will be transmitted to the terminal.

[0068] In this embodiment, regardless of whether nucleic acid is present in the sample solution, the electrode current will increase from the initial value during the detection process. However, under the same conditions, the increase in electrode current when nucleic acid is present in the sample solution will be less than the increase in electrode current when nucleic acid is not present in the sample solution.

[0069] In the embodiments of this application, when nucleic acid is present in the sample solution, the change in electrode current will become smaller and smaller as the concentration of nucleic acid increases during the detection process.

[0070] In this embodiment, the electrode disposed on the bottom wall of the third reaction chamber 63 is an OECTs (Organic Electrochemical Transistor) electrode. The OECTs electrode is prepared as follows: a PET (polyethylene terephthalate) substrate (0.188 mm thick, Grafix DURA-RAR) is subjected to plasma treatment for 5 minutes and then cleaned with isopropanol. Subsequently, a photoresist (BP212-37S) is uniformly spin-coated onto the PET substrate, exposed and developed under ultraviolet light, and a 10 nm Cr electrode layer and a 90 nm Au electrode layer are sequentially deposited by thermal evaporation. Next, an organic channel layer containing PEDOT:PSS is prepared on the substrate using a spin-coating process. The PEDOT:PSS solution (containing 0.1 wt% sodium dodecylbenzenesulfonate, 5 wt% ethylene glycol, and 1 wt% GOPS) for the OECTs channel layer is spin-coated onto the photoresist pattern. Subsequently, it is annealed at 100°C for 30 minutes to optimize the film performance. Finally, SU-8 photoresist is used to form a protective insulating layer to protect the electrodes.

[0071] The automated amplified nucleic acid detection device provided in this application has excellent sensitivity, good specificity and wide applicability, making highly sensitive automated nucleic acid molecular detection possible.

[0072] It should be noted that in this application, the terms "upper," "lower," "left," "right," "front," "rear," "top," "bottom," "inner," "outer," "vertical," "horizontal," "lateral," and "longitudinal" indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. These terms are mainly for the purpose of better describing this application and its embodiments, and are not intended to limit the indicated device, element, or component to having a specific orientation, or to be constructed and operated in a specific orientation.

[0073] Furthermore, in addition to indicating location or positional relationship, some of the aforementioned terms may also have other meanings. For example, the term "above" may also be used in some cases to indicate a certain dependency or connection relationship. Those skilled in the art can understand the specific meaning of these terms in this application based on the specific circumstances.

[0074] Furthermore, the terms "installation," "setup," "equipped with," "connection," and "linked" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral structure; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium, or an internal connection between two devices, components, or parts. Those skilled in the art can understand the specific meaning of these terms in this application based on the specific circumstances.

[0075] Furthermore, the terms "first," "second," etc., are primarily used to distinguish different devices, elements, or components (which may be the same or different in specific type and construction), and are not intended to indicate or imply the relative importance or quantity of the indicated devices, elements, or components. Unless otherwise stated, "a plurality of" means two or more.

[0076] The above description is merely a specific embodiment of the present invention, enabling those skilled in the art to understand or implement the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the present invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features claimed herein.

Claims

1. An automated amplification-free nucleic acid detection device, characterized in that, include: Installation components are used to provide installation space; The first fluid input component is used to input the sample solution; The first fluid input component is disposed on the mounting component; A second fluid input component is used to input the detection reagent solution; the second fluid input component is disposed on the mounting component. The third fluid input component is used to input the flushing solution; The third fluid input component is disposed on the mounting component; A fourth fluid input component is used to input a protein streptavidin solution; the fourth fluid input component is disposed on the mounting component. A detection component is used to detect the presence of nucleic acids in a sample solution; the detection component is disposed on the mounting component and is connected to the first fluid input component, the second fluid input component, the third fluid input component, and the fourth fluid input component, respectively; A fluid output component for outputting the post-reaction mixed solution; the fluid output component is disposed on the mounting component and connected to the detection component; A control component is used to control the various parts; the control component is disposed on the mounting component and is connected to the first fluid input component, the second fluid input component, the third fluid input component, the fourth fluid input component, the detection component, and the fluid output component, respectively.

2. The automated amplification-free nucleic acid detection device according to claim 1, characterized in that, The mounting assembly includes a housing and a mounting plate, wherein the mounting plate is disposed inside the housing, and the first fluid input component, the second fluid input component, the third fluid input component, the fourth fluid input component, the detection component, the fluid output component, and the control component are respectively disposed on the mounting plate.

3. The automated amplification-free nucleic acid detection device according to claim 1, characterized in that, The first fluid input component includes a first micropump and a first fluid delivery pipe, wherein the first micropump is connected to the control component and the first fluid delivery pipe, and the first fluid delivery pipe is connected to the detection component.

4. The automated amplification-free nucleic acid detection device according to claim 1, characterized in that, The second fluid input component includes a second micropump and a second fluid delivery pipe, wherein the second micropump is connected to the control component and the second fluid delivery pipe, respectively, and the second fluid delivery pipe is connected to the detection component.

5. The automated amplification-free nucleic acid detection device according to claim 1, characterized in that, The third fluid input component includes a third micro pump and a third fluid delivery pipe, wherein the third micro pump is connected to the control component and the third fluid delivery pipe respectively, and the third fluid delivery pipe is connected to the detection component.

6. The automated amplification-free nucleic acid detection device according to claim 1, characterized in that, The fourth fluid input component includes a fourth micropump and a fourth fluid delivery pipe, wherein the fourth micropump is connected to the control component and the fourth fluid delivery pipe respectively, and the fourth fluid delivery pipe is connected to the detection component.

7. The automated amplification-free nucleic acid detection device according to claim 1, characterized in that, The detection assembly includes a first reaction chamber, a second reaction chamber, and a third reaction chamber. The first reaction chamber is connected to a first fluid delivery pipe in the first fluid input assembly, a second fluid delivery pipe in the second fluid input assembly, a fourth fluid delivery pipe in the fourth fluid input assembly, and the second reaction chamber. The third reaction chamber is connected to a third fluid delivery pipe in the third fluid input assembly, the fluid output assembly, and the second reaction chamber. A heater is provided on the bottom wall of the first reaction chamber and is connected to the control assembly. An electrode is provided on the bottom wall of the third reaction chamber and is connected to the control assembly.

8. The automated amplification-free nucleic acid detection device according to claim 1, characterized in that, The fluid output assembly includes a fluid output pipe, wherein the fluid output pipe is connected to the third reaction chamber in the detection assembly.

9. The automated amplification-free nucleic acid detection device according to claim 1, characterized in that, The control component includes a microfluidic control module, a heating controller, a micropump controller, and a wireless transmission module. The microfluidic control module is connected to the heating controller and the micropump controller. The heating controller is connected to the heater in the detection component. The micropump controller is connected to the first micropump in the first fluid input component, the second micropump in the second fluid input component, the third micropump in the third fluid input component, and the fourth micropump in the fourth fluid input component. The wireless transmission module is connected to the electrodes in the detection component.

10. The automated amplification-free nucleic acid detection device according to claim 9, characterized in that, The control component further includes a transmission end, which is connected to the wireless transmission module and the electrode respectively.

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