Magnetic aggregation induced emission particle fluorescence-electrochemical nucleic acid dual-detection chip and detection system

Abstract

The invention discloses a magnetic aggregation induced emission particle fluorescence-electrochemical nucleic acid dual detection chip and system.The chip comprises a chip body and an upper cover, and a plurality of electrochemical reaction detection cavities and fluorescence reaction detection cavities are formed in the left side and the right side of the chip body in the front-back direction respectively; the electrochemical reaction detection cavities and the fluorescence reaction detection cavities are arranged in a one-to-one correspondence manner; each electrochemical reaction detection cavity and each fluorescence reaction detection cavity are provided with a flow channel, and one end of each flow channel is communicated with the electrochemical reaction detection cavity or the fluorescence reaction detection cavity; the other end of the sampling tube is communicated with an MAIE electrochemical detection auxiliary liquid sample adding hole and an MAIE fluorescence detection auxiliary liquid sample adding hole which are formed in the chip main body; a plurality of MAIE electrochemical detection sample adding holes and MAIE fluorescence detection sample adding holes are formed in the upper cover, and corresponding openings are formed in the positions, corresponding to the MAIE electrochemical detection auxiliary liquid adding holes and the MAIE fluorescence detection auxiliary liquid adding holes, of the upper cover.

Description

Technical Field

[0001] The present invention belongs to the field of biological detection technology, and in particular relates to a magnetic aggregation-induced emission particle (MAIE) fluorescence-electrochemical nucleic acid dual detection chip and a detection system. Background Art

[0002] Nucleic acid detection technology plays a crucial role in the field of molecular diagnostics. However, existing single-electrochemical nucleic acid detection methods often suffer from reduced accuracy due to environmental interference and nonspecific reactions, resulting in false positive or false negative results, seriously affecting the reliability of test results.

[0003] Multi-system detection is achieved by combining two or more detection methods, such as the combination of electrochemistry and surface enhanced Raman scattering (SERS), or the combination of fluorescence and colorimetric detection, to achieve multiple signal outputs for the same detection target. The advantage of this multi-system detection method is that different detection methods do not interfere with each other and can verify each other, thereby significantly improving the reliability of detection. It is particularly suitable for solving problems such as detection of low-concentration markers, non-specific reactions and environmental interference. However, there are also some defects. For example, electrochemical-SERS dual-mode detection requires the simultaneous integration of an electrochemical workstation and a Raman spectrometer, which has high requirements for equipment synchronization and signal acquisition timing, is difficult to integrate, and is cumbersome to operate. In addition, electrochemical processes (such as potential scanning) may cause changes in the microenvironment of the electrode surface (such as pH fluctuations, bubble generation), resulting in SERS signal drift or distortion, poor signal synchronization, and electrochemical reactions may destroy the uniformity of the SERS substrate layer structure, reducing signal reproducibility; electrochemical side reactions will interfere with the recognition of SERS spectra, and impurities in complex samples (such as blood, sewage) may contaminate the SERS substrate or clog the electrode, reducing long-term stability, all of which limit its practical application scenarios.

[0004] In summary, there is an urgent need to provide a new type of multi-system detection platform that is simple to operate, stable, not restricted by scenarios, and can simultaneously achieve sensitive, rapid and efficient detection of multiple target substances, providing innovative technical solutions for scientific research and applications in related fields. Summary of the Invention

[0005] To address the above-mentioned technical problems existing in existing electrochemical detection technology, the present invention provides a magnetic aggregation-induced emission particle (MAIE) fluorescence-electrochemical nucleic acid dual detection chip and detection system, which enables synchronous monitoring of electrochemical and fluorescence signals of samples within the chip. It can be heated at variable temperatures and incubated at different temperatures in the electrochemical reaction detection chamber and the fluorescence reaction detection chamber. It is easy to operate and miniaturized.

[0006] The technical solution adopted in the present invention is:

[0007] The first aspect of the present invention relates to a magnetic aggregation-induced luminescence microparticle fluorescence-electrochemical nucleic acid dual detection chip, comprising a chip body (9), a temperature control device (8) being provided at the bottom of the chip body (9), and an upper cover (1) being provided above the chip body (9), wherein the length direction of the chip body (9) is defined as the front-to-back direction, and the width direction is defined as the left-to-right direction; wherein:

[0008] A plurality of electrochemical reaction detection cavities (6) and fluorescence reaction detection cavities (7) are respectively provided on the left and right sides of the chip body (9) along the front-back direction, and the electrochemical reaction detection cavities (6) and the fluorescence reaction detection cavities (7) are arranged one by one opposite to each other; each of the electrochemical reaction detection cavities (6) and the fluorescence reaction detection cavities (7) is provided with a flow channel (10), and one end of each of the flow channels (10) is connected to the electrochemical reaction detection cavity (6) or the fluorescence reaction detection cavity (7); and the other end is connected to the MAIE electrochemical detection auxiliary liquid sample addition hole (11) and the MAIE fluorescence detection auxiliary liquid sample addition hole (12) provided on the chip body (9);

[0009] The upper cover (1) is provided with a plurality of MAIE electrochemical detection sample loading holes (13) and MAIE fluorescence detection sample loading holes (14), and the MAIE electrochemical detection sample loading holes (13) and MAIE fluorescence detection sample loading holes (14) respectively correspond to the positions of the electrochemical reaction detection chamber (6) and the fluorescence reaction detection chamber (7); and the upper cover (1) is provided with openings for the MAIE electrochemical detection auxiliary liquid loading holes (11) and openings for the MAIE fluorescence detection auxiliary liquid loading holes (12) at positions corresponding to the MAIE electrochemical detection auxiliary liquid loading holes (11) and the MAIE fluorescence detection auxiliary liquid loading holes (12).

[0010] Preferably, a CRISPR / Cas reaction element that reacts with the nucleic acid to be detected is provided in the electrochemical reaction detection chamber (6), and the bottom of the electrochemical reaction detection chamber (6) has a first open end;

[0011] A fluorescent reaction element that reacts with the nucleic acid to be detected is provided in the fluorescent reaction detection chamber (7), and the bottom of the fluorescent reaction detection chamber (7) has a second open end;

[0012] The first opening end and the second opening end are both provided with a three-electrode system.

[0013] Preferably, the three-electrode system comprises an auxiliary electrode (2), a working electrode (3) and a reference electrode (4); the auxiliary electrode (2) and the reference electrode (4) are respectively arranged around the working electrode (3); the auxiliary electrode (2), the working electrode (3) and the reference electrode (4) are respectively connected to electrode contacts (5); and the electrode contacts (5) are used to be electrically connected to an external supporting instrument.

[0014] Preferably, the temperature control device (8) is composed of a heating resistor.

[0015] Preferably, the flow channel (10), the electrochemical reaction detection chamber (6), and the fluorescence reaction detection chamber (7) are distributed on the same plane.

[0016] Preferably, temperature measurement points are provided in both the electrochemical reaction detection chamber (6) and the fluorescence reaction detection chamber (7), and the temperature is fed back through the temperature measurement points to control the temperature of the electrochemical reaction detection chamber (6) and the fluorescence reaction detection chamber (7).

[0017] Preferably, the electrochemical reaction detection chamber (6) contains a biotinylated CRISPR / Cas reaction element coated with MAIE, a MAIE fluorescent reporter probe and a MAIE electrochemical reporter probe.

[0018] Preferably, the fluorescent reaction detection cavity (7) has a fluorescent reaction element coated with a fluorescent reporter group.

[0019] A second aspect of the present invention relates to a magnetic particle luminescent double-layer microfluidic detection system, the detection system comprising:

[0020] The magnetic particle luminescent double-layer microfluidic chip as described above;

[0021] A magnet unit, used for driving the magnetic aggregation-induced emission particles (MAIE) in the MAIE primer solution to move;

[0022] An extrusion unit, used for squeezing the labeled primer storage portion and the cleaning solution storage portion to allow the labeled ligand and the cleaning solution to flow out;

[0023] The detection unit is used to detect the luminescence and electrical signals in the detection area.

[0024] Compared with the prior art, the beneficial effects of the present invention are embodied in:

[0025] (1) The present invention encapsulates biotinylated CRISPR / Cas reaction components, MAIE fluorescent reporter probes and MAIE electrochemical reporter probes, in the reaction detection chamber, enabling simultaneous monitoring of electrochemical and fluorescent signals, and enabling sensitive, rapid, and efficient detection of multiple target substances simultaneously.

[0026] (2) The present invention can be heated at variable temperatures, and incubation at different temperatures can be performed in the electrochemical reaction detection chamber and the fluorescence reaction detection chamber, which is easy to operate and miniaturized.

[0027] (3) The combination of electrochemical and fluorescence detection technologies in the present invention fully utilizes the complementary advantages of the two: electrochemical direct measurement of current or potential changes can reflect reaction information in real time and quantitatively, and is suitable for high-sensitivity, rapid, and real-time dynamic monitoring; while fluorescence detection reduces background interference and enhances specificity and sensitivity through the specific binding of target molecules with fluorescent probes (such as aptamers and antibodies). The combination of the two can significantly improve the sensitivity and accuracy of detection. In addition, the dual-mode detection of electrochemical and fluorescence detection has strong resistance to environmental interference, is insensitive to solution turbidity and color, and is suitable for complex samples (such as blood and sewage).

[0028] (4) The synergistic effect of the electrochemical and fluorescence detection technologies of the present invention is excellent. Fluorescence provides molecular recognition signals, electrochemistry verifies reaction activity, and cross-validation can reduce the risk of false positives / negatives. It has unique advantages, especially in multiple analysis, trace substance detection and complex sample analysis. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 This is a top view of the flow channel chamber component provided by the present invention (excluding the upper cover).

[0030] Figure 2 This is a schematic diagram of the layered structure of the MAIE fluorescence-electrochemical nucleic acid dual detection chip provided by the present invention.

[0031] Figure 3 This is a structural diagram of the MAIE fluorescence-electrochemical nucleic acid dual detection chip provided by the present invention (including the upper cover). DETAILED DESCRIPTION

[0032] The specific implementation of the new embodiment of the present invention is described in detail below with reference to the accompanying drawings. It should be understood that the specific implementation described herein is only used to illustrate and explain the new embodiment of the present invention, and is not used to limit the new embodiment of the present invention.

[0033] In the description of the present invention, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", "clockwise", "counterclockwise", "axial", "radial", "circumferential", etc., indicating the orientation or positional relationship, are based on the orientation or positional relationship shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operate in a specific orientation, and therefore should not be understood as limiting the present invention. In addition, the embodiments of the present invention and the features therein may be combined with each other unless there is a conflict.

[0034] The terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of the technical features being referred to. Thus, a feature specified as "first" or "second" may explicitly or implicitly include at least one such feature. In the description of the present invention, "plurality" means at least two, for example, two, three, etc., unless otherwise specifically defined.

[0035] The present invention designs a magnetic aggregation-induced emission particle (MAIE) fluorescence-electrochemical nucleic acid dual detection chip and detection system. In order to make the purpose, technical solution and advantages of the present invention clearer, the present invention will be described in detail with reference to the accompanying drawings and in combination with exemplary embodiments.

[0036] Example 1

[0037] refer to Figure 1 、 Figure 2 and Figure 3 The present invention provides a magnetic aggregation-induced luminescence microparticle fluorescence-electrochemical nucleic acid dual detection chip, comprising a chip body 9, a temperature control device 8 being provided at the bottom of the chip body 9, and an upper cover 1 being provided above the chip body 9. The length direction of the chip body 9 is defined as the front-to-back direction, and the width direction is defined as the left-to-right direction; wherein:

[0038] A plurality of electrochemical reaction detection cavities 6 and fluorescent reaction detection cavities 7 are respectively provided on the left and right sides of the chip body 9 along the front-to-back direction. The electrochemical reaction detection cavities 6 and fluorescent reaction detection cavities 7 are arranged one by one opposite to each other. Each of the electrochemical reaction detection cavities 6 and fluorescent reaction detection cavities 7 is provided with a flow channel 10. One end of each flow channel 10 is connected to the electrochemical reaction detection cavity 6 or the fluorescent reaction detection cavity 7; the other end is connected to the MAIE electrochemical detection auxiliary liquid addition hole 11 and the MAIE fluorescence detection auxiliary liquid addition hole 12 provided on the chip body 9.

[0039] The upper cover 1 is provided with a plurality of MAIE electrochemical detection sample addition holes 13 and MAIE fluorescence detection sample addition holes 14, and the MAIE electrochemical detection sample addition holes 13 and MAIE fluorescence detection sample addition holes 14 correspond to the positions of the electrochemical reaction detection chamber 6 and the fluorescence reaction detection chamber 7 respectively; the upper cover 1 is provided with openings for the MAIE electrochemical detection auxiliary liquid addition hole 11 and the MAIE fluorescence detection auxiliary liquid addition hole 12 at positions corresponding to the MAIE electrochemical detection auxiliary liquid addition hole 11 and the MAIE fluorescence detection auxiliary liquid addition hole 12.

[0040] In this embodiment, a CRISPR / Cas reaction element that reacts with the nucleic acid to be detected is provided in the electrochemical reaction detection chamber 6, and the bottom of the electrochemical reaction detection chamber 6 has a first open end;

[0041] The fluorescent reaction detection chamber 7 is provided with a fluorescent reaction element that reacts with the nucleic acid to be detected, and the bottom of the fluorescent reaction detection chamber 7 has a second open end;

[0042] The first opening end and the second opening end are both provided with a three-electrode system.

[0043] In this embodiment, the three-electrode system includes an auxiliary electrode 2, a working electrode 3 and a reference electrode 4; the auxiliary electrode 2 and the reference electrode 4 are respectively surrounded by the working electrode 3, and the auxiliary electrode 2, the working electrode 3 and the reference electrode 4 are respectively connected to the electrode contacts 5, and the electrode contacts 5 are used to be electrically connected to external supporting instruments.

[0044] In this embodiment, the temperature control device 8 is composed of a heating resistor.

[0045] In this embodiment, the flow channel 10 , the electrochemical reaction detection chamber 6 , and the fluorescence reaction detection chamber 7 are distributed on the same plane.

[0046] In this embodiment, temperature measurement points are provided in the electrochemical reaction detection chamber 6 and the fluorescence reaction detection chamber 7 , and the temperature is fed back through the temperature measurement points to control the temperature of the electrochemical reaction detection chamber 6 and the fluorescence reaction detection chamber 7 .

[0047] In this embodiment, the electrochemical reaction detection chamber 6 contains a biotinylated CRISPR / Cas reaction element coated with MAIE, a MAIE fluorescent reporter probe, and a MAIE electrochemical reporter probe.

[0048] Specifically, the electrochemical reaction detection chamber 6 is provided with a biotinylated crRNA (targeting the SARS-CoV-2 target gene) fixed on the surface of the working electrode to be tested, a CRISPR / Cas12a complex pre-assembled with the Cas12a protein, and a methylene blue (MB) reporter probe reaction part labeled at both ends of ssDNA.

[0049] In this embodiment, the fluorescent reaction detection cavity 7 contains a fluorescent reaction element coated with a fluorescent reporter group.

[0050] Specifically, the fluorescence reaction detection chamber 7 is provided with a MAIE-ssDNA fluorescent reporter probe that reacts with the same crRNA-Cas12a complex as the working electrode surface fixed in the above-mentioned electrochemical reaction detection chamber 6 and the SARS-CoV-2 nucleic acid to be tested.

[0051] Example 2

[0052] A magnetic particle luminescent double-layer microfluidic detection system of the present invention comprises:

[0053] The magnetic particle luminescent double-layer microfluidic chip as described above;

[0054] A magnet unit, used for driving the magnetic aggregation-induced emission particles (MAIE) in the MAIE primer solution to move;

[0055] An extrusion unit, used for squeezing the labeled primer storage portion and the cleaning solution storage portion to allow the labeled ligand and the cleaning solution to flow out;

[0056] The detection unit is used to detect the luminescence and electrical signals in the detection area.

[0057] In this embodiment, the cleaning solution is used to clean the MAIE microparticles to remove non-specifically adsorbed analytes, luminescent markers, and other substances that may affect the detection results.

[0058] Specifically, the cleaning solution can be a buffer reagent, protein and surfactant, wherein the buffer reagent includes but is not limited to phosphate, Tris-HCl, borate, and acetate, the protein includes but is not limited to bovine serum albumin, casein, etc., and the surfactant includes but is not limited to polyvinyl pyrrolidone, polyethylene glycol, Triton X-100, Tween, etc.; the pH range of the cleaning solution is 6.0 to 10.0.

[0059] More specifically, the washing solution used was a pH 7.4 Tris-HCl buffer containing bovine serum albumin.

[0060] When conducting a test in the present invention, the operator adds 10 to 30 μL of the nucleic acid sample to be tested into the MAIE electrochemical detection sample addition well 13 and the MAIE fluorescence detection sample addition well 14 using a syringe, a pipette, or other tools, and adds 5 to 10 μL of an adapted auxiliary liquid into the corresponding MAIE electrochemical detection auxiliary liquid addition well 11 and the MAIE fluorescence detection auxiliary liquid addition well 12, and incubates at an adapted temperature.

[0061] In the electrochemical reaction detection chamber 6, when the liquid completely covers the three-electrode system, if the nucleic acid sample to be tested contains target RNA, the target RNA can form a ternary complex with the CRISPR / Cas protein and CRISPR / crRNA. This ternary complex activates the trans-cleavage activity of the CRISPR / Cas protein, resulting in the RNA chain in the reporter molecule being sheared. This process will cause the number of complexes to decrease, thereby changing the resistance of the electrode surface. By manipulating the instrument to apply voltage to the electrode contact 5, the reduction of the complex on the electrode surface will lead to a decrease in resistance, thereby causing an increase in the electrochemical signal. Therefore, when the target RNA is present in the nucleic acid sample to be tested, the current on the electrode will increase, which is manifested as an increase in the electrochemical detection signal. On the contrary, when the nucleic acid sample to be tested does not contain target RNA, the trans-cleavage activity of the CRISPR / Cas protein will not be activated, the RNA chain in the reporter molecule will not be sheared, and the ternary complex remains unchanged. At this time, the resistance of the electrode surface will increase, resulting in a decrease in current, thereby reducing the electrochemical detection signal. Therefore, when there is no target RNA in the nucleic acid sample to be tested, the electrochemical detection signal will weaken.

[0062] In the fluorescence reaction detection chamber 7, when the liquid completely covers the three-electrode system, if the sample to be tested contains target RNA, the target RNA can form a ternary complex with the CRISPR / Cas protein and CRISPR / crRNA, thereby activating the trans-cleavage activity of the CRISPR / Cas protein. At this time, the RNA chain in the reporter molecule is sheared, releasing aggregation-induced emission (AIE) fluorescent groups. The released fluorescent groups will diffuse to the electrode surface and participate in the electrochemical reaction, causing the fluorescent groups to be excited, thereby emitting a fluorescent signal. By applying voltage to the electrode contacts 5, the electrochemical reaction occurring on the electrode surface will prompt more fluorescent groups to participate in the excitation, further enhancing the fluorescence signal. At the same time, AIE causes fluorescence aggregation, and the fluorescence signal is significantly enhanced. Therefore, when the target RNA is present in the sample to be tested, the fluorescence signal will be enhanced. On the contrary, if the sample to be tested does not contain target RNA, the trans-cleavage activity of the CRISPR / Cas protein will not be activated, the RNA chain in the reporter molecule will not be sheared, and the fluorescent group cannot be released. Therefore, no additional fluorescent groups will be produced on the electrode surface, and the fluorescence signal will be weakened. In other words, when there is no target RNA in the sample to be tested, the enhancement of the fluorescence signal caused by fluorescence quenching due to AIE will be significantly reduced.

[0063] Example 3

[0064] refer to Figure 1, dual-color MAIE microparticles based on different fluorescence wavelengths are used to detect different targets of the same nucleic acid: In this embodiment three, two types of HPV, HPV16 and HPV18, are taken as examples. The electrochemical reaction detection chamber 6 is provided with a biotinylated crRNA (targeting the HPV16 target gene) fixed on the surface of the working electrode to be tested, and a CRISPR / Cas12a complex pre-assembled with the Cas12a protein and a MAIE-ssDNA (480nm) fluorescent reporter probe reaction piece that reacts with the HPV16 nucleic acid to be tested; the fluorescent reaction detection chamber 7 is provided with a MAIE-ssDNA (620nm) fluorescent reporter probe that reacts with the same crRNA-Cas12a complex as the working electrode surface fixed in the above-mentioned electrochemical reaction detection chamber 6 and the HPV18 nucleic acid to be tested. The detection chamber 6 and the detection chamber 7 can both be electrochemical detection chambers or fluorescent detection chambers, and the corresponding electrochemical detection sample loading hole 13 and the MAIE fluorescent detection sample loading hole 14 can be used interchangeably.

[0065] Compared with the existing methods based on a single detection mode, this chip achieves the simultaneous detection of nucleic acids and other target substances on the same platform by combining magnetic aggregation-induced luminescence and electrochemical and fluorescence detection technologies. This gives the chip significant advantages in the detection of multiple biomarkers and has excellent specificity and sensitivity, making it suitable for multiple fields such as clinical diagnosis, environmental monitoring, and food safety.

[0066] Although the embodiments of the novel invention have been shown and described above, it is understood that the above embodiments are exemplary and are not to be construed as limitations on the novel invention. A person skilled in the art may change, modify, replace and modify the above embodiments within the scope of the novel invention.

Claims

1. A magnetic aggregation-induced luminescence microparticle fluorescence-electrochemical nucleic acid dual detection chip, comprising a chip body (9), a temperature control device (8) provided at the bottom of the chip body (9), an upper cover (1) provided above the chip body (9), wherein the length direction of the chip body (9) is defined as the front-to-back direction, and the width direction is defined as the left-to-right direction; and characterized in that: A plurality of electrochemical reaction detection cavities (6) and fluorescence reaction detection cavities (7) are respectively provided on the left and right sides of the chip body (9) along the front-back direction, and the electrochemical reaction detection cavities (6) and the fluorescence reaction detection cavities (7) are arranged one by one opposite to each other; each of the electrochemical reaction detection cavities (6) and the fluorescence reaction detection cavities (7) is provided with a flow channel (10), and one end of each of the flow channels (10) is connected to the electrochemical reaction detection cavity (6) or the fluorescence reaction detection cavity (7); and the other end is connected to the MAIE electrochemical detection auxiliary liquid sample addition hole (11) and the MAIE fluorescence detection auxiliary liquid sample addition hole (12) provided on the chip body (9); The upper cover (1) is provided with a plurality of MAIE electrochemical detection sample loading holes (13) and MAIE fluorescence detection sample loading holes (14), and the MAIE electrochemical detection sample loading holes (13) and MAIE fluorescence detection sample loading holes (14) respectively correspond to the positions of the electrochemical reaction detection chamber (6) and the fluorescence reaction detection chamber (7); and the upper cover (1) is provided with openings for the MAIE electrochemical detection auxiliary liquid loading holes (11) and openings for the MAIE fluorescence detection auxiliary liquid loading holes (12) at positions corresponding to the MAIE electrochemical detection auxiliary liquid loading holes (11) and the MAIE fluorescence detection auxiliary liquid loading holes (12).

2. The magnetic aggregation-induced luminescence microparticle fluorescence-electrochemical nucleic acid dual detection chip according to claim 1, characterized in that: The electrochemical reaction detection chamber (6) is provided with a CRISPR / Cas reaction element that reacts with the nucleic acid to be detected, and the bottom of the electrochemical reaction detection chamber (6) has a first open end; A fluorescent reaction element that reacts with the nucleic acid to be detected is provided in the fluorescent reaction detection chamber (7), and the bottom of the fluorescent reaction detection chamber (7) has a second open end; The first opening end and the second opening end are both provided with a three-electrode system.

3. The magnetic aggregation-induced luminescence microparticle fluorescence-electrochemical nucleic acid dual detection chip according to claim 2, characterized in that: The three-electrode system comprises an auxiliary electrode (2), a working electrode (3) and a reference electrode (4); the auxiliary electrode (2) and the reference electrode (4) respectively surround the working electrode (3); the auxiliary electrode (2), the working electrode (3) and the reference electrode (4) are respectively connected to electrode contacts (5); and the electrode contacts (5) are used to be electrically connected to an external supporting instrument.

4. The magnetic aggregation-induced luminescence microparticle fluorescence-electrochemical nucleic acid dual detection chip according to claim 1, characterized in that: The temperature control device (8) is composed of a heating resistor.

5. The magnetic aggregation-induced luminescence microparticle fluorescence-electrochemical nucleic acid dual detection chip according to claim 1, characterized in that: The flow channel (10), the electrochemical reaction detection chamber (6), and the fluorescence reaction detection chamber (7) are distributed on the same plane.

6. The magnetic aggregation-induced luminescence microparticle fluorescence-electrochemical nucleic acid dual detection chip according to claim 1, characterized in that: Temperature measurement points are provided in both the electrochemical reaction detection chamber (6) and the fluorescence reaction detection chamber (7), and the temperature is fed back through the temperature measurement points to control the temperature of the electrochemical reaction detection chamber (6) and the fluorescence reaction detection chamber (7).

7. The magnetic aggregation-induced luminescence microparticle fluorescence-electrochemical nucleic acid dual detection chip according to claim 3, characterized in that: The electrochemical reaction detection chamber (6) contains a biotinylated CRISPR / Cas reaction element coated with MAIE, a MAIE fluorescent reporter probe, and a MAIE electrochemical reporter probe.

8. The magnetic aggregation-induced luminescence microparticle fluorescence-electrochemical nucleic acid dual detection chip according to claim 3, characterized in that: The fluorescent reaction detection cavity (7) contains a fluorescent reaction element coated with a fluorescent reporter group.

9. A magnetic particle luminescent double-layer microfluidic detection system, characterized in that: The detection system comprises: The magnetic particle luminescent double-layer microfluidic chip according to any one of claims 1 to 8; A magnet unit, used for driving the magnetic aggregation-induced emission particles (MAIE) in the MAIE primer solution to move; An extrusion unit, used for squeezing the labeled primer storage portion and the cleaning solution storage portion to allow the labeled ligand and the cleaning solution to flow out; The detection unit is used to detect the luminescence and electrical signals in the detection area.

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