Rapid detection chip for high-sensitivity cardiac troponin and its detection method

Abstract

The present invention discloses a high-sensitivity cardiac troponin rapid detection chip and a detection method thereof; the high-sensitivity cardiac troponin rapid detection chip includes a substrate having a blood collection port penetrating through the top surface and the bottom surface, and a plurality of microchannels are provided on the top surface of the substrate, each of which is centered on the blood collection port and gradually spirally expands outward. Each of the microchannels is sequentially divided into a red blood cell treatment area, an incubation area, and a detection area from the inner circle to the outer circle. Immobilized biomolecules or biomolecular films capable of retaining red blood cells or rupturing red blood cells are respectively provided in each section of the red blood cell treatment area, and the functions of each section of the incubation area and the detection area are regulated by method one or method two; the present invention adopts microfluidic technology, which can solve the environmental and time-effect problems in the blood sample collection and its processing process of the existing human cardiac troponin detection, and the prepared microfluidic rapid detection chip for human high-sensitivity cardiac troponin is small in size and low in preparation cost.

Description

Technical Field

[0001] The present invention relates to the technical field of cardiac troponin detection, in particular to the technical field of high-sensitivity cardiac troponin detection chips. Background Art

[0002] Acute coronary syndrome (ACS) is a group of clinical syndromes based on the pathological basis of rupture or invasion of coronary atherosclerotic plaques and subsequent formation of complete or incomplete occlusive thrombi, including acute ST-segment elevation myocardial infarction, acute non-ST-segment elevation myocardial infarction, and unstable angina (UA); ACS often presents with symptoms such as episodic chest pain and chest tightness, which can lead to arrhythmia and heart failure (even sudden death), seriously affecting the quality of life and lifespan of its patients; most ACS patients delay the timely diagnosis and treatment of ACS due to lack of sufficient attention to mild chest tightness and chest pain symptoms (if appropriate treatment methods are taken in a timely manner for ACS patients, the mortality rate can be greatly reduced, complications can be reduced, and the prognosis can be improved).

[0003] Currently, the diagnosis, prognosis assessment, treatment, and risk stratification of ACS are mainly judged based on the content of cardiac troponin (cTn) in human blood; cTn is a regulatory protein for myocardial muscle contraction, consisting of subunits of three different genes, including cardiac troponin T (cTnT), cardiac troponin I (cTnI), and troponin C (TnC); among them, both cTnI and cTnT are encoded by separate genes and are biomarkers of myocardial injury; cTnI mainly participates in the inhibition of myocardial muscle contraction, with strong specificity, generally increasing gradually in peripheral blood 4 - 8 hours after myocardial injury and reaching the highest value 12 - 24 hours after myocardial injury (elevated cTnI can still be detected in peripheral blood 7 - 10 days after myocardial injury); cTnT plays a connecting role in regulating muscle contraction, and like cTnI, is mainly expressed in the myocardium and can also be detected in small amounts in skeletal muscle, but has a longer half-life and exists in the blood for a longer time (this makes cTnT suitable for observing later-stage conditions); in addition, in addition to being used in the diagnosis and treatment of acute myocardial injury, cTnT detection can also be used for risk stratification of patients with chronic myocardial diseases (such as heart failure); that is to say, rapid, sensitive, and accurate determination of single cTnI, cTnT in blood or simultaneous detection of both is of great significance for the diagnosis of acute myocardial infarction, risk stratification of acute coronary syndrome, monitoring of myocardial injury caused by various factors, and prognosis judgment of cardiac events.

[0004] At present, the main detection methods for cTnI and / or cTnT are enzyme-linked immunosorbent assay (ELISA) and chemiluminescence assay. Among them, for the ELISA method, such as the triple-antibody method for rapid quantitative determination of cardiac troponin I with the publication number CN1271409C, there are usually problems of long operation time and low automation degree. For the chemiluminescence assay, such as the method for detecting cardiac troponin I / T by flash homogeneous chemiluminescence technology with the publication number CN110031635A, although it has the characteristics of strong specificity, high sensitivity and high accuracy, it often requires expensive instrument equipment and experienced operators, and can basically only be used in specific medical institutions.

[0005] With the continuous improvement of the detection sensitivity of cardiac troponin and the wide development of the detection of high-sensitivity cardiac troponin in clinical laboratories, it is urgent to solve the problem of how to avoid drawing venous blood (that is, without the operation of professional medical staff) and reduce the detection time (the detection time of existing methods is usually more than about 10 minutes) on the basis of realizing point-of-care testing (POCT), so as to achieve on-site testing for individuals at home or grass-roots medical staff. Summary of the Invention

[0006] The purpose of the present invention is to solve the problems in the prior art, and propose a rapid detection chip for high-sensitivity cardiac troponin and its detection method. By adopting an integrated microfluidic technology, it can solve the environmental and time-effect problems in the blood sample collection and its processing process of the existing human cardiac troponin detection, and the prepared microfluidic rapid detection chip for human high-sensitivity cardiac troponin is small in size and low in preparation cost.

[0007] To achieve the above object, the present invention provides a high-sensitivity cardiac troponin rapid detection chip, comprising a substrate having a blood collection port penetrating through the top surface and the bottom surface. The substrate is provided with a plurality of microchannels on the top surface, each of which is centered on the blood collection port and gradually spirally expands outward. Each microchannel is sequentially divided into a red blood cell processing area, an incubation area, and a detection area from the inner circle to the outer circle. Immobilized biomolecules or biomolecular films capable of retaining red blood cells or rupturing red blood cells are respectively provided in each section of the red blood cell processing area. The functions of each section of the incubation area and the detection area are regulated by method one or method two. Method one is to respectively provide a conjugate formed by coupling a colloidal carrier and a cardiac troponin-specific polyclonal antibody only in each section of the incubation area. Method two is to respectively provide a cardiac troponin-specific first monoclonal antibody (abbreviation: first antibody) labeled with a fluorescent substance in each section of the incubation area on the one hand, and on the other hand, to respectively provide a cardiac troponin-specific second monoclonal antibody (abbreviation: second antibody) capable of capturing an immune complex formed by the binding of the cardiac troponin-specific first monoclonal antibody labeled with a fluorescent substance and cardiac troponin in each detection area; Microfluidics is a new technology that has emerged in recent years, which refers to using channels with a size of dozens to hundreds of micrometers to process or manipulate extremely small amounts (10 -9 ~10 -18 liters) of liquid, and one of its important characteristics is the extremely strong surface tension at the micrometer scale, having unique fluid properties such as laminar flow, vortex flow, and droplets, etc.; In the present invention, by setting each microchannel as a gradually expanding spiral shape, the blood can exhibit a strong secondary vortex (i.e., the Dean effect) during the flowing process, thereby realizing the transport dynamics of trace amounts of blood that are difficult to complete by a series of conventional methods, effectively improving the interaction between the blood and the substances provided in each section of the red blood cell processing area and the incubation area (realizing some unconventional high-speed chemical reactions), shortening the blood flow time and reducing the chip size at the same time; In addition, the number of microchannels provided can be designed according to the required detection quantity of high-sensitivity cardiac troponin (such as only detecting cTnI, only detecting cTnT, or simultaneously detecting cTnI and cTnT).

[0008] Preferably, the number of turns of each section of the red blood cell processing area is 1 to 2 turns, the number of turns of each section of the incubation area is 2 to 10 turns, and each microchannel forms a detection cavity with a relatively large size at the tail end as the detection area.

[0009] The Dean number is an important indicator for measuring the size of secondary eddies in each of the described microchannels, which depends on properties such as the bend curvature, channel dimensions, fluid inertial effects, and viscous effects; as the Dean number increases, the secondary eddies in the channel become stronger, causing multiple exchanges and contacts between the blood and the channel surface and triggering strong surface catalytic reactions; based on this, in the present invention, the hydraulic diameter of each section of the red blood cell processing area is 10-100 μm, the hydraulic diameter of each section of the incubation area is 5-100 μm, the diameter of each of the detection chambers is 20-500 μm and the depth is 10-500 μm; during use, when blood enters each microchannel from the blood collection port, it will first pass through each section of the red blood cell processing area to rupture or intercept the red blood cells and allow the remaining serum / plasma to continue to move deeper; since each section of the red blood cell processing area is located in the innermost part of the respective microchannel, the channel curvature is the largest in the whole channel and the Dean effect is the strongest; for this reason, the present invention reduces the blood flow rate by increasing the hydraulic diameter of each section of the red blood cell processing area, thereby balancing the excessive cell rupture that may be caused by the Dean flow effect; the second part of each microchannel (i.e., each section of the incubation area) is respectively used for the formation of immune complexes (when blood passes through, cTnI and / or cTnT therein can respectively bind to their specific antibodies to form immune complexes); to ensure that the immune complexes can fully complete the incubation reaction, the present invention increases the Dean effect by relatively reducing the hydraulic diameter of this part of the channel to obtain a larger flow rate; in addition, since the present invention sets a relatively large number of spiral turns for each section of the incubation area, it can further ensure that the immune incubation reaction is fully completed; that is to say, the present invention can balance the differences in the incubation reaction rates of different protein subunits by adjusting the hydraulic diameter of different sections of each microchannel.

[0010] Preferably, the cross-sections of each section of the red blood cell processing area and the incubation area are square, circular, trapezoidal or triangular.

[0011] Preferably, each section of the red blood cell processing area uses a hemolytic agent to rupture red blood cells or an anti-red blood cell antibody to intercept red blood cells.

[0012] Furthermore, the hemolytic agent is one or a combination of cetyltrimethylammonium chloride (CTAB), polysorbate, and saponin.

[0013] Furthermore, the anti-red blood cell antibody is anti-red blood cell membrane glycoprotein A (anti-CD235a) and / or anti-hemoglobin antibody (Anti-Hb).

[0014] Preferably, the colloidal carrier is latex particles.

[0015] Preferably, the fluorescent substance is fluorescein and / or fluorescent microspheres containing fluorescein.

[0016] Preferably, the troponin-specific polyclonal antibody is a polyclonal antibody that can recognize cTnI and / or cTnT, and both the troponin-specific first monoclonal antibody and the troponin-specific second monoclonal antibody are monoclonal antibodies that can recognize cTnI and / or cTnT; in addition, when selecting troponin-specific antibodies, attention needs to be paid to their cross-reactivity (i.e., cross-reactivity between the three subunits of cTn should be avoided) to ensure high specificity in detection.

[0017] Furthermore, the monoclonal antibody that can recognize cTnI is a specific amino acid sequence at the N-terminus or C-terminus of cTnI (such as cTnI-2, etc.).

[0018] Furthermore, the polyclonal antibody that can recognize cTnI is a goat anti-cTnI polyclonal antibody, a rabbit anti-cTnI polyclonal antibody, or a specific amino acid sequence in the N-terminal region, central region, or C-terminal region of different target epitopes of cTnI (such as residues 1-30, 31-110, or 111-181, etc.).

[0019] Furthermore, the monoclonal antibody that can recognize cTnT is a C-terminal epitope of cTnT (such as cTnL-2, etc.).

[0020] Furthermore, the polyclonal antibody that can recognize cTnT is a rabbit-derived polyclonal antibody (such as Ab188879 or Ab64750, etc.), a rabbit anti-cTnT polyclonal antibody (such as PA5-14314, etc.), or a polyclonal antibody SAB1405743.

[0021] The troponin-specific first monoclonal antibody labeled with a fluorescent substance is usually a troponin monoclonal antibody labeled with a fluorescein (such as FITC and Alexa Fluor, etc.), a fluorescent microsphere of the fluorescein, or other probes, to amplify the signal or perform fluorescence detection; at the same time, the correct selection and use of the second antibody are crucial for the successful detection of the immune complex formed by the troponin-specific antibody labeled with a fluorescent substance and troponin; specifically, in the detection of cTnI, the first antibody against the N-terminal epitope (such as residues 1-30) is used to recognize the N-terminal epitope and conjugate with the fluorescein, and the second antibody is used to recognize the central region or C-terminal epitope of cTnI, such as a monoclonal antibody against -cTnI (residues 50-110); the first antibody against the C-terminal epitope (such as residues 150-181) is used to recognize the C-terminal epitope and conjugate with the fluorescent microsphere, and the second antibody is used to recognize the N-terminal or central region of the cTnI antibody, such as a monoclonal antibody against -cTnI (residues 1-49); in addition, in the detection of cTnT, the first antibody against the N-terminal epitope (such as residues 1-80) is used to target the N-terminal epitope and conjugate with the fluorescein, and the second antibody is used to recognize the central or C-terminal epitope, such as a monoclonal antibody against -cTnT (residues 100-150), and the first antibody against the C-terminal epitope (such as residues 160-290) is used to recognize the C-terminal epitope and conjugate with the fluorescent microsphere, and the second antibody is used to recognize the N-terminal or central region, such as a monoclonal antibody against -cTnT (residues 1-50).

[0022] Preferably, the top surface of the substrate is covered with a cover plate that is transparent and can seal the blood collection port and each microchannel, and a sticker that can seal the blood collection port is attached to the bottom surface of the substrate. The cover plate and the sticker jointly seal the blood collection port and each microchannel and form a vacuum environment isolated from the outside or a closed environment filled with a non-oxygen gas.

[0023] Furthermore, the thicknesses of both the substrate and the cover plate are 100 - 1000 μm.

[0024] Furthermore, the substrate is made of a polymer material, a pure metal material, an alloy material, or a non-metal material.

[0025] Still further, the polymer material is PDMS or polystyrene, the pure metal material is aluminum or copper, and the non-metal material is a carbon-based material or a silicon-based material.

[0026] Furthermore, the cover plate is a transparent inorganic glass plate, a transparent organic glass plate, or an optical organic film.

[0027] Furthermore, the non-oxygen gas is an inert gas.

[0028] Still further, the inert gas is one or a combination of two of nitrogen and argon.

[0029] Furthermore, optical anti-reflection coatings are respectively provided on the front and back surfaces of the cover plate, and optical reflection coatings are respectively provided on each section of the incubation area and the detection area; wherein, each section of the incubation area and the detection area can respectively utilize the optical reflection coating to increase the optical signal intensity, while the cover plate can reduce the reflection of the detected incident light and improve the transmittance of the blood sample optical signal through the optical anti-reflection coatings on its front and back surfaces.

[0030] Still further, the optical anti-reflection coating is a metal thin film (such as silver or aluminum, etc.).

[0031] Still further, the optical reflection coating is a TiN thin film; the TiN thin film has a very high light reflectivity in the infrared region. In particular, this thin film not only has high density, low manufacturing cost and biocompatibility, but also has a mature process in integrated circuit production.

[0032] Still further, the surfaces of each section of the incubation area and the detection area need to be chemically polished or chemically-mechanically polished first before setting the optical reflection coating until the surface roughness is less than 0.5 μm (the specific roughness may depend on the wavelength of the detected light); wherein, the chemical polishing or chemically-mechanically polishing treatment can further enhance the light reflectivity of each section of the incubation area and the detection area.

[0033] The present invention also provides a detection method, which needs to use the above-mentioned high-sensitivity cardiac troponin rapid detection chip and the specific operation steps are as follows: first, send blood from the bottom surface of the substrate into the blood collection port, and then when the blood flows to each detection area (the immune complexes that have undergone sufficient incubation reaction will finally enter the terminals of each microchannel), respectively perform quantitative analysis of the fluorescence intensity (fluorescence method) or solution turbidity (turbidimetry) on each detection area; wherein, cTnI or cTnL with known concentration can be used for calibration and establish a standard curve; when in use, compare the measurement data with the standard curve to determine the concentration of cTnI and / or cTnL.

[0034] Preferably, for the fluorescence method, waste liquid areas can be respectively added at the terminals of each microchannel to collect the liquid that has not formed the fluorescent complex.

[0035] Preferably, a separated spectrophotometer penetrates the cover plate at a wavelength of 250-900 nm and detects the fluorescence intensity or solution turbidity reflected by each detection area; wherein, the measurement of the reflected light can effectively increase the measurement accuracy; in addition, the specific fluorescence wavelength depends on the excitation and emission characteristics of the fluorescent substance, such as the commonly used organic fluorescein FITC (excitation wavelength is about 495 nm and emission wavelength is about 519 nm) or rhodamine fluorescein (excitation wavelength is about 540 nm and emission wavelength is about 570 nm) and lanthanide elements for time-resolved fluorescence (such as Eu3+ The excitation wavelength is about 395 nm and the emission wavelength is about 610 nm).

[0036] Preferably, the dot matrix light spectroscopy measurement technology is used to separately perform point-by-point scanning measurement on each section of the incubation area along the blood flow direction to obtain the real-time reaction progress of the immune incubation reaction along the channel flow direction (such a process can further improve the sensitivity of chip measurement on the one hand, and can also assist in parameter selection during early development on the other hand, achieving the effects of optimizing specific antibodies and optimizing the channel length).

[0037] Advantages of the present invention:

[0038] 1) The integrated microfluidic chip formed by integrating the blood collection port and several multi-stage micron-scale channels with detection cavities in the present invention, and biofilms with different functions are coated in sections along the blood flow direction in each integrated microchannel. Thus, blood can be directly drawn from the fingertip end of the human body during testing, and the blood is fully mixed and reacted with the corresponding biofilm in each microchannel before flowing into the detection cavity. Finally, the micro-concentration determination of cardiac troponin is realized through quantitative analysis of fluorescence intensity or solution turbidity. Moreover, the whole process does not require professional medical staff to assist in blood extraction and processing, solving the environmental and time-effect problems of existing cardiac troponin detection, providing the possibility of rapid detection for personalized and clinical doctor's precision medicine, and can be manufactured by using the integrated production processes and equipment commonly used in micron-scale large-scale integrated circuit production. The preparation process is simple, mature and stable, and the preparation cost is low, meeting the expenditure requirements of ordinary people for daily medical consumables;

[0039] 2) By setting each microchannel in a mosquito coil shape in the present invention, the vortex effect of the fluid in each microchannel can be effectively enhanced, thereby shortening the length of each microchannel. At the same time, its circular structure can also greatly reduce the chip size, and thus correspondingly reduce the production cost of the chip;

[0040] 3) By respectively adding optical reflection coatings in each section of the incubation area and the detection area in the present invention, the integrated microfluidic chip can capture relatively weak spectral signals;

[0041] 4) By designing different hydraulic diameters for each section of the red blood cell treatment area and the incubation area in the present invention, the processing requirements for different functions (red blood cell treatment and incubation) can be met;

[0042] 5) By adding a transparent encapsulation cover plate on the substrate, the present invention can realize the optical detection of substances in the detection cavity by selecting the material of the transparent encapsulation cover plate. Moreover, a vacuum environment isolated from the outside or a closed environment filled with non-oxygen gas can be formed between the substrate and the transparent encapsulation cover plate, thereby avoiding the contact of trace human blood with the atmosphere and ensuring the accuracy of detection.

[0043] 6) By adopting the dot matrix light spectrum measurement technology, the present invention separately performs point-by-point scanning measurement on each incubation area along the blood flow direction, so as to facilitate obtaining in real time the reaction progress of each incubation area along the channel (which in turn facilitates the developer to optimize the channel length).

[0044] The features and advantages of the present invention will be described in detail through embodiments in conjunction with the accompanying drawings. Description of the Drawings

[0045] Figure 1 is the three-dimensional structure schematic diagram of Embodiment 1;

[0046] Figure 2 is the top view of Embodiment 1;

[0047] Figure 3 is Figure 2 the sectional view taken along the line A-A of

[0048] Figure 4 is the physical diagram of Embodiment 1;

[0049] Figure 5 is the three-dimensional structure schematic diagram of Embodiment 2;

[0050] Figure 6 is the top view of Embodiment 2;

[0051] Figure 7 is Figure 6 the sectional view taken along the line B-B of

[0052] In the figures: 1 - substrate, 11 - blood collection port, 12 - microchannel, 121 - detection cavity, 2 - cover plate. Detailed Description of the Specific Embodiment

[0053] Embodiment 1:

[0054] Refer to Figures 1 to 4, this embodiment provides a high-sensitivity cardiac troponin rapid detection chip, including a substrate 1 with a blood collection port 11 penetrating the top and bottom surfaces. Two microchannels 12 are provided on the top surface of the substrate 1, each centered on the blood collection port 11 and gradually spiraling outward. Each microchannel 12 is sequentially divided into a red blood cell treatment area, an incubation area, and a detection area from the inner circle to the outer circle. Immobilized biomolecules or biomolecular films capable of retaining red blood cells or rupturing red blood cells are respectively provided in each section of the red blood cell treatment area. The function of each section of the incubation area and the detection area is regulated by method one or method two. Method one is to only provide a conjugate formed by coupling a colloidal carrier with a cardiac troponin-specific polyclonal antibody in each section of the incubation area. Method two is to provide a fluorescently labeled cardiac troponin-specific first monoclonal antibody in each section of the incubation area on the one hand and a cardiac troponin-specific second monoclonal antibody capable of capturing the immune complex formed by the binding of the fluorescently labeled cardiac troponin-specific first monoclonal antibody and cardiac troponin in each detection area on the other hand.

[0055] The number of turns of each section of the red blood cell treatment area is 1 turn, the number of turns of each section of the incubation area is 3 turns, and each microchannel 12 forms a detection cavity 121 with a relatively large size at the tail end as the detection area.

[0056] The hydraulic diameter of each section of the red blood cell treatment area is 10 - 100 μm, the hydraulic diameter of each section of the incubation area is 5 - 100 μm, the diameter of each detection cavity 121 is 20 - 500 μm, and the depth is 10 - 500 μm.

[0057] The cross-sections of each section of the red blood cell treatment area and the incubation area are trapezoidal.

[0058] Each section of the red blood cell treatment area uses a hemolytic agent to rupture red blood cells or an anti-red blood cell antibody to retain red blood cells.

[0059] The cardiac troponin-specific polyclonal antibody is a polyclonal antibody capable of recognizing cTnI and / or cTnT. Both the cardiac troponin-specific first monoclonal antibody and the cardiac troponin-specific second monoclonal antibody are monoclonal antibodies capable of recognizing cTnI and / or cTnT.

[0060] The top surface of the substrate 1 is covered with a cover plate 2 that is transparent and can seal the blood collection port 11 and each microchannel 12. A sticker that can seal the blood collection port 11 is attached to the bottom surface of the substrate 1. The cover plate 2 and the sticker jointly seal the blood collection port 11 and each microchannel 12 and form a vacuum environment isolated from the outside or a closed environment filled with a non-oxygen gas.

[0061] The front and back sides of the cover plate 2 are respectively provided with an optical anti-reflection coating, and each section of the incubation area and the detection area is respectively provided with an optical reflection coating.

[0062] This embodiment also provides a detection method, which uses the above-mentioned high-sensitivity cardiac troponin rapid detection chip, and the specific operation steps are as follows: first, blood is fed into the blood collection port 11 from the bottom surface of the substrate 1, and then when the blood flows to each detection area, quantitative analysis of the fluorescence intensity or solution turbidity is carried out on each detection area respectively.

[0063] The dot matrix optical spectrum measurement technology is used to separately perform point-by-point scanning measurement on each section of the incubation area along the blood flow direction to obtain the real-time reaction progress of the immune incubation reaction along the channel flow direction.

[0064] Embodiment 2:

[0065] Refer to Figures 5 to 7 , for the high-sensitivity cardiac troponin rapid detection chip of the present invention, the substrate 1 is provided with four micro-channels 12 on the top surface, which are respectively centered on the blood collection port 11 and gradually spiral outward.

[0066] The cross-sections of each section of the red blood cell treatment area and the incubation area are all circular.

[0067] Others are the same as those in Embodiment 1.

[0068] The above embodiments are descriptions of the present invention, not limitations on the present invention. Any solution obtained by simply transforming the present invention belongs to the protection scope of the present invention.

Claims

1. High-sensitivity cardiac troponin rapid detection chip, characterized by: The invention comprises a substrate (1) having a blood collection port (11) penetrating the top and bottom surfaces, wherein the substrate (1) is provided with a plurality of microchannels (12) on the top surface, each of which is centered on the blood collection port (11) and gradually expands in a spiral manner toward the periphery, wherein each of the microchannels (12) is divided into a red blood cell processing area, an incubation area, and a detection area in sequence from the inner circle to the outer circle, wherein each section of the red blood cell processing area is provided with an immobilized biomolecule or biomolecule film capable of intercepting or rupturing red blood cells, and wherein the functions of the incubation area and the detection area of ​​each section are regulated by the first method or the second method, wherein the first method is to only provide a combination formed by coupling a colloidal carrier with a troponin-specific polyclonal antibody in each section of the incubation area, and the second method is to provide a troponin-specific first monoclonal antibody labeled with a fluorescent substance in each section of the incubation area, and to provide a troponin-specific second monoclonal antibody capable of capturing an immune complex formed by the combination of the troponin-specific first monoclonal antibody labeled with a fluorescent substance and troponin in each detection area; The number of turns of each section of the red blood cell treatment zone is 1 to 2 turns, the number of turns of each section of the incubation zone is 2 to 10 turns, the hydraulic diameter of each section of the red blood cell treatment zone is 10 to 100 μm, and the hydraulic diameter of each section of the incubation zone is 5 to 100 μm; The top surface of the substrate (1) is covered with a transparent cover plate (2) that can seal the blood collection port (11) and each microchannel (12); the front and back surfaces of the cover plate (2) are respectively provided with optical anti-reflection coatings; each section of the incubation area and the detection area is respectively provided with an optical reflective coating; a separate spectrophotometer penetrates the cover plate (2) and detects the fluorescence intensity or solution turbidity reflected by each detection area; and the surfaces of each section of the incubation area and the detection area are first subjected to chemical polishing or chemical mechanical polishing before the optical reflective coating is provided.

2. The high-sensitivity cardiac troponin rapid detection chip according to claim 1, characterized in that: Each of the microchannels (12) forms a relatively large detection cavity (121) at the tail end to serve as a detection area.

3. The high-sensitivity cardiac troponin rapid detection chip as claimed in claim 2, characterized in that: The diameter of each detection cavity (121) is 20 to 500 μm and the depth is 10 to 500 μm.

4. The high-sensitivity cardiac troponin rapid detection chip according to claim 1, characterized in that: The cross-sections of the red blood cell processing area and the incubation area in each section are square, circular, trapezoidal or triangular.

5. The high-sensitivity cardiac troponin rapid detection chip according to claim 1, characterized in that: The red blood cell processing area described in each section uses a hemolytic agent to rupture the red blood cells or uses an anti-red blood cell antibody to retain the red blood cells.

6. The high-sensitivity cardiac troponin rapid detection chip according to claim 1, characterized in that: The troponin-specific polyclonal antibody is a polyclonal antibody that can recognize cTnI and / or cTnT, and the troponin-specific first monoclonal antibody and the troponin-specific second monoclonal antibody are both monoclonal antibodies that can recognize cTnI and / or cTnT.

7. The high-sensitivity cardiac troponin rapid detection chip according to claim 1, characterized in that: The bottom surface of the substrate (1) is affixed with a sticker capable of sealing the blood collection port (11), and the cover plate (2) and the sticker together seal the blood collection port (11) and each microchannel (12) and form a vacuum environment isolated from the outside or a closed environment filled with non-oxygen gas.

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