Blood coagulation function and thrombus marker combined detection platform and method

Abstract

The invention discloses a blood coagulation function and thrombus marker combined detection platform and method, and belongs to the field of blood detection.The detection device comprises a static magnetic field generation structure, a detection coil, a heating assembly and a magnetoelastic sensor, the heating assembly is used for heating a blood sample, and the magnetoelastic sensor is located in the center of the detection coil; the detection coil is electrically connected with the impedance analysis device, current is applied to the detection coil to generate an alternating magnetic field for working of the magnetoelastic sensor, the static magnetic field generation structure is used for generating a static magnetic field for working of the magnetoelastic sensor, and magnetic lines of the static magnetic field are parallel to magnetic lines of the alternating magnetic field. The impedance analysis device drives the detection coil to generate an alternating magnetic field around the magnetoelastic sensor, the magnetoelastic sensor generates resonance in the alternating magnetic field, the detection coil generates an induction signal, the impedance analysis device collects the induction signal, and through the design, joint detection of the thrombus marker and the blood coagulation function can be achieved on a single platform.

Description

Technical Field

[0001] The present invention relates to the field of blood testing, and in particular to a platform and method for combined detection of coagulation function and thrombosis markers. Background Art

[0002] Epidemiological studies have shown that thrombotic diseases account for 51% of deaths worldwide, far exceeding any other disease. Disseminated intravascular coagulation (DIC), a serious thrombotic disease, has a mortality rate as high as 80%. However, clinically, there is a lack of clear indicators for early diagnosis of thrombosis, and with the absence of obvious clinical manifestations, the probability of early detection is only 1%. Early identification and correction of coagulopathy is crucial for reducing transfusion requirements, reducing complications, and improving survival rates.

[0003] Currently, commonly used clinical thrombosis molecular marker tests, such as D-dimer, fibrin / fibrinogen degradation products (FDPs), and prothrombin activation inhibitor (PAI-1), can more sensitively and reliably reflect the occurrence and formation of thrombus, pre-DIC status, and vascular endothelial damage in the early stages. However, these marker tests can usually only provide fragmented information and cannot fully reflect the dynamic changes in the coagulation process. Thromboelastography (TEG), as a real-time detection method that reflects the dynamic changes in blood coagulation (including fibrinolysis), can provide parameters such as coagulation initiation time, coagulation formation rate, and clot stability, which helps to assess dynamic coagulation function. However, TEG mainly reflects the interaction between blood cells and coagulation factors and cannot directly reflect the damage to the vascular endothelium. Clinical studies have shown that the combined use of thrombosis marker detection and thromboelastography can significantly improve the diagnostic accuracy of thrombosis and DIC.

[0004] Thrombosis marker detection methods primarily include enzyme-linked immunosorbent assay (ELISA), immunoturbidimetry, chemiluminescence immunoassay, and fluorescence immunoassay. Thromboelastography methods primarily include the hanging wire method, resonant frequency method, rotational method, and ultrasonic resonance method. Despite the diverse development of existing coagulation testing methods, these methods generally suffer from limitations such as long testing cycles, complex procedures, and high sample volume requirements. Furthermore, thrombus marker detection and thrombus detection require separate equipment, resulting in lengthy testing cycles and high blood volume requirements. Summary of the Invention

[0005] In order to overcome the deficiencies of the prior art, one of the objectives of the present invention is to provide a detection platform that can realize the combined detection of thrombosis markers and coagulation function on a single platform.

[0006] In order to overcome the deficiencies of the prior art, a second object of the present invention is to provide a detection method that can realize the combined detection of thrombosis markers and coagulation function on a single platform.

[0007] One of the purposes of the present invention is achieved by the following technical solution:

[0008] A combined detection platform for coagulation function and thrombus markers includes a detection device, a temperature control device, and an impedance analysis device. The detection device includes a static magnetic field generating structure, a detection coil, a heating component, and a magnetoelastic sensor. The heating component is used to heat a blood sample. The magnetoelastic sensor is located at the center of the detection coil. The detection coil is electrically connected to the impedance analysis device. A current is applied to the detection coil to generate an alternating magnetic field for the operation of the magnetoelastic sensor. The static magnetic field generating structure is used to generate a static magnetic field for the operation of the magnetoelastic sensor. The magnetic lines of force of the static magnetic field are parallel to the magnetic lines of force of the alternating magnetic field. The impedance analysis device drives the detection coil to generate the alternating magnetic field around the magnetoelastic sensor. The magnetoelastic sensor resonates in the alternating magnetic field. The detection coil generates an induction signal, and the impedance analysis device collects the induction signal.

[0009] Furthermore, the static magnetic field generating structure is a permanent magnet or a Helmholtz coil.

[0010] Furthermore, when the detection device performs thrombus marker detection, the surface of the magnetoelastic sensor is modified with specific antibodies.

[0011] Furthermore, when the detection device performs a coagulation function test, the magnetoelastic sensor is located in the blood that is in the coagulation process.

[0012] Furthermore, the heating assembly includes a heat-conducting column and a heating belt wound around the heat-conducting column, and the magnetoelastic sensor is located in the heat-conducting column.

[0013] Furthermore, the detection device further includes a mounting base, which is detachably mounted inside the heat-conducting column and is used to carry the magnetoelastic sensor and the blood sample.

[0014] Furthermore, the heating component is located in the detection coil.

[0015] Furthermore, the heating belt is a copper belt wrapped with polyimide.

[0016] Furthermore, there are two detection devices, each of which is connected to the temperature control device and the impedance analysis device respectively, and the two detection devices perform coagulation function detection and thrombus marker detection on the blood sample respectively.

[0017] Furthermore, the number of the detection device is one, and the detection device performs coagulation function detection and thrombosis marker detection on the blood sample at different times.

[0018] The second object of the present invention is achieved by adopting the following technical solution:

[0019] A detection method based on the above-mentioned coagulation function and thrombosis marker combined detection platform comprises the following steps:

[0020] Thrombus marker concentration detection: a magnetoelastic sensor modified with a specific antibody is prepared and placed inside a detection coil of a detection device; a static magnetic field generating structure generates a static magnetic field for the magnetoelastic sensor to operate, and the detection coil generates an alternating magnetic field around the magnetoelastic sensor. The magnetic lines of force of the static magnetic field are parallel to the magnetic lines of force of the alternating magnetic field, and the magnetoelastic sensor resonates in the alternating magnetic field. The impedance analysis device collects the electrical properties of the detection coil and records the initial resonant frequency of the magnetoelastic sensor; a blood sample is added so that the magnetoelastic sensor modified with the specific antibody is located in the blood sample. The specific antibodies modified on the surface of the magnetoelastic sensor capture the thrombus marker molecules in the sample being tested, causing the load mass on the sensor surface to change, thereby changing the resonant frequency of the sensor. The impedance analysis device collects the electrical properties of the detection coil and records the response resonant frequency of the magnetoelastic sensor; the offset of the resonant frequency is calculated, and a calibration curve is obtained based on the resonant frequency offset corresponding to different thrombus marker concentrations, and the thrombus marker concentration in the sample being tested is calculated;

[0021] Coagulation function test: A blood sample and an activator are prepared in proportion, and the mixed sample is added so that the magnetoelastic sensor is located in the mixed sample, and the mixed sample is placed inside the detection coil of the detection device; the static magnetic field generating structure generates a static magnetic field for the operation of the magnetoelastic sensor, and the detection coil generates an alternating magnetic field around the magnetoelastic sensor. The magnetic lines of force of the static magnetic field are parallel to the magnetic lines of force of the alternating magnetic field, and the magnetoelastic sensor resonates in the alternating magnetic field. When the blood coagulates under the action of the activator, the viscoelasticity of the blood will continue to change. When the magnetoelastic sensor works in the continuously coagulating blood, due to the viscous damping effect, the energy generated by the magnetoelastic sensor in the resonant state will be lost, and the impedance amplitude will decrease. The impedance analysis device collects the electrical characteristics of the detection coil, records the impedance amplitude of the magnetoelastic sensor, obtains a time-impedance amplitude change curve, and extracts coagulation-related parameters.

[0022] Furthermore, the thrombus marker concentration detection step and the coagulation function detection step are performed separately on the two detection devices.

[0023] Furthermore, the thrombus marker concentration detection step and the coagulation function detection step are performed on a detection device.

[0024] Compared with the existing technology, the combined detection platform for coagulation function and thrombus markers of the present invention includes a detection device, a temperature control device and an impedance analysis device. The detection device includes a static magnetic field generating structure, a detection coil, a heating component and a magnetoelastic sensor. The heating component is used to heat the blood sample. The magnetoelastic sensor is located at the center of the detection coil. The detection coil is electrically connected to the impedance analysis device. The detection coil is applied with current to generate an alternating magnetic field for the operation of the magnetoelastic sensor. The static magnetic field generating structure is used to generate a static magnetic field for the operation of the magnetoelastic sensor. The magnetic lines of force of the static magnetic field are parallel to the magnetic lines of force of the alternating magnetic field. The impedance analysis device drives the detection coil to generate an alternating magnetic field around the magnetoelastic sensor. The magnetoelastic sensor resonates in the alternating magnetic field. The detection coil generates an induction signal. The impedance analysis device collects the induction signal. Through the above design, the combined detection of thrombus markers and coagulation function can be realized on a single platform. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 This is a schematic structural diagram of the first embodiment of the combined detection platform for coagulation function and thrombosis markers of the present invention;

[0026] Figure 2 for Figure 1 A partial structural diagram of the detection device of the coagulation function and thrombosis marker combined detection platform;

[0027] Figure 3 This is a schematic structural diagram of a second embodiment of the combined detection platform for coagulation function and thrombosis markers of the present invention;

[0028] Figure 4 is the impedance amplitude-frequency characteristic curve of the magnetoelastic sensor;

[0029] Figure 5 Schematic diagram of the surface modification process of the magnetoelastic sensor;

[0030] Figure 6 This is a schematic diagram of thrombus marker concentration detection;

[0031] Figure 7 It is the calibration curve of thrombus marker concentration;

[0032] Figure 8 This is a schematic diagram of the coagulation function test principle;

[0033] Figure 9 It is the time-impedance amplitude change curve of blood coagulation process;

[0034] Figure 10 The figure is a flow chart of the combined detection method of coagulation function and thrombosis markers of the present invention.

[0035] In the figure: 10, detection device; 11, static magnetic field generating structure; 12, detection coil; 13, heating component; 130, thermal conductive column; 131, heating belt; 14, mounting base; 141, plate; 142, sample slot; 15, magnetoelastic sensor; 20, temperature control device; 30, impedance analysis device; 40, processor. DETAILED DESCRIPTION

[0036] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. All other embodiments obtained by ordinary technicians in this field based on the embodiments of the present invention without making any creative efforts shall fall within the scope of protection of the present invention.

[0037] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention pertains. The terms used in this specification of the present invention are for the purpose of describing specific embodiments only and are not intended to limit the present invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0038] See also Figure 1 as well as Figure 2 , which is the first embodiment of the combined detection platform for coagulation function and thrombosis markers of the present invention, the combined detection platform for coagulation function and thrombosis markers includes a detection device 10, a temperature control device 20, an impedance analysis device 30 and a processor 40.

[0039] In this embodiment, two detection devices 10 are provided, one for coagulation function testing and the other for thrombus marker detection. Specifically, the detection device 10 includes a static magnetic field generating structure 11, a detection coil 12, a heating assembly 13, a mounting base 14, and a magnetoelastic sensor 15. The static magnetic field generating structure 11 is used to generate a static magnetic field for the magnetoelastic sensor 15 to operate. The static magnetic field generating structure 11 can be a permanent magnet or a Helmholtz coil.

[0040] When the static magnetic field generating structure is a permanent magnet, the permanent magnet is cylindrical or rectangular, and the material includes but is not limited to nepheline iron boron, ferrite, etc., and the magnetized end surface area is about 3-4cm 2 The thickness range is 2-5mm, the magnetization direction is the thickness direction, and the surface magnetic field range is 500-2500G. The permanent magnet is parallel to the central axis of the detection coil 12, that is, the magnetic lines of force of the static magnetic field are parallel to the magnetic lines of force of the alternating magnetic field.

[0041] Impedance analyzer 30 applies an alternating current to detection coil 12, generating an alternating magnetic field around magnetoelastic sensor 15. Detection coil 12 is wound with enameled wire. Specifically, in this embodiment, detection coil 12 is wound with enameled wire having a wire diameter of 0.2-0.4 mm. It can be cylindrical in shape, with a diameter of 15-20 mm, a length of 30-40 mm, and 150-200 turns. Detection coil 12 should be wound as evenly and smoothly as possible, and can be wound in multiple layers.

[0042] The heating assembly 13 provides the required temperature environment for sample detection and includes a heat-conducting column 130 and a heating belt 131. The heating assembly 13 is used to heat the sample slot 142 area of the mounting base 14, where the sample and the magnetoelastic sensor 15 are placed.

[0043] In this embodiment, the heat-conducting column 130 is made of a ceramic material, such as silicon nitride, aluminum oxide, etc., which has the characteristics of insulation, heat conduction and non-magnetic conductivity. The heat-conducting column 130 is cylindrical in shape, 30 mm in length and 15-20 mm in diameter, with a square through groove inside for placing the mounting seat 14 and forming a tight wrap around it. The heating belt 131 is a thin belt made of a copper belt wrapped with polyimide. It is evenly and symmetrically wrapped around the ceramic surface to reduce electromagnetic influence. It can cause heat by applying current and heat the inside of the ceramic by heat conduction. In addition, the heating belt 131 can also be contained inside the ceramic.

[0044] Mounting base 14 is inserted and installed inside thermally conductive pillar 130 and is located at the center of detection coil 12. Mounting base 14 includes a plate 141 with a sample slot 142 for accommodating the sample and magnetoelastic sensor 15. Sample slot 142 is 4.3 mm wide, 10.3 mm long, and 0.2-0.4 mm deep.

[0045] The magnetization state of the magnetoelastic sensor 15 changes under magnetic field excitation. Due to the law of electromagnetic induction, the induced voltage or impedance of the detection coil 12 will change accordingly. This induced voltage or impedance characterizes the electrical characteristics of the magnetoelastic sensor 15. The electrical signal can be collected by the impedance analysis device 30. The processor and the impedance analysis device 30 can measure and record the impedance amplitude-frequency characteristic curve of the magnetoelastic sensor 15 in real time, as well as the resonant frequency and resonant impedance amplitude contained in the curve, such as Figure 4 In this example, the resonant frequency is defined as the frequency corresponding to the maximum impedance amplitude in the impedance amplitude-frequency characteristic curve of the magnetoelastic sensor, and the resonant impedance amplitude is defined as the maximum impedance amplitude in the impedance amplitude-frequency characteristic curve of the magnetoelastic sensor.

[0046] Specifically, the magnetoelastic sensor 15 is made of an amorphous soft magnetic alloy material, and its composition is usually a combination of two or more of iron, cobalt, nickel, molybdenum, silicon, boron, copper, and niobium, such as iron silicon boron, iron nickel molybdenum boron, iron nickel boron copper, cobalt iron nickel silicon boron, iron niobium copper silicon boron, etc. Amorphous soft magnetic alloy materials have magnetostrictive properties, that is, they will undergo mechanical deformation under the action of an external magnetic field. When the excitation frequency of the magnetic field is close to the intrinsic frequency of the sensor, the sensor will resonate, and the vibration process will change the magnetization state in the sensor material, thereby changing the magnetic flux density of the sensor. Amorphous soft magnetic alloys are cut into different shapes through different processes to prepare sensors, including rectangles, triangles, sectors, etc. The present invention prepares the material into a thin sheet with a length of 10 mm and a width of 4 mm by laser cutting, and the thickness is usually 20-50 μm.

[0047] The temperature control device 20 is electrically connected to the heating belt 131 and has power regulation and temperature feedback measurement functions to achieve temperature control of the heating component. This module is realized through circuit design and development.

[0048] The impedance analysis device 30 is electrically connected to the detection coil 12. The impedance analysis device 30 has the following functions: first, an excitation output function, which generates an AC voltage or current signal of a specific frequency and amplitude, used to drive the detection coil 12 to generate an alternating magnetic field around the magnetoelastic sensor 15; second, real-time acquisition of the induced voltage or impedance change on the detection coil 12, which reflects the resonant characteristics of the magnetoelastic sensor 15. Third, the acquired signal is transmitted to an external storage device or data analysis system for further processing and recording. The impedance analysis device 30 can be implemented using commercial impedance analysis equipment, such as an LCR meter or impedance analyzer, or through circuit design and development.

[0049] The processor 40 is electrically connected or in communication connection with the impedance analysis device 30 , and processes the collected signals.

[0050] When the coagulation function and thrombus marker combined detection platform of the present application is installed, the permanent magnet is parallel to the central axis of the detection coil 12, that is, the magnetic lines of force of the static magnetic field are parallel to the magnetic lines of force of the alternating magnetic field; the distance between the end face of the permanent magnet in the magnetization direction and the center of the coil is in the range of 3-6 cm, and the heating component 13, the mounting base 14, and the magnetoelastic sensor 15 are placed inside the detection coil 12 from the outside to the inside. The heating component 13 is electrically connected to the temperature control device 20 by means of leads; the detection coil 12 receives the magnetic field changes of the magnetoelastic sensor 15 wirelessly and is electrically connected to the impedance analysis device 30 by means of leads; the impedance analysis device 30 communicates with the host computer via a signal transmission line, such as USB, RS232, GPIB, etc.

[0051] Please continue reading Figure 3This is the second embodiment of the combined coagulation function and thrombosis marker detection platform of the present invention. In this second embodiment, the structure of the combined coagulation function and thrombosis marker detection platform is substantially the same as that of the first embodiment, with the difference being that there is only one detection device 10, and both coagulation function and thrombosis marker detection are performed in the same detection device 10. The difference between the coagulation function and thrombosis marker detection lies in the sample and magnetoelastic sensor 15. The thrombosis marker detection sample includes a thrombosis marker calibration sample with a concentration gradient range and a blood sample with an unknown thrombosis marker concentration. The magnetoelastic sensor 15 is a magnetoelastic sensor modified with a specific antibody. The coagulation function detection sample is a mixed sample formed by mixing a whole blood sample and an activator in a certain proportion. The magnetoelastic sensor 15 does not require modification.

[0052] Please continue reading Figure 10 The present application also discloses a method for combined detection of coagulation function and thrombosis markers based on the above-mentioned combined detection platform, comprising the following steps:

[0053] Thrombus marker concentration detection steps and coagulation function detection steps.

[0054] The principle of thrombus marker concentration detection is as follows: the magnetoelastic sensor 15 detects thrombus marker concentration based on the mass loading effect. When there is no mass loading on the surface of the magnetoelastic sensor 15, its resonant frequency is its own unloaded characteristic frequency. When there is a certain mass loading on the surface of the magnetoelastic sensor 15, its resonant frequency shifts. By modifying the surface of the magnetoelastic sensor 15 with specific antibodies to capture thrombus marker molecules (antigens), the mass loading on the surface of the magnetoelastic sensor 15 changes, which in turn causes the resonant frequency of the magnetoelastic sensor 15 to change. By establishing a calibration curve between the resonant frequency offset of the magnetoelastic sensor 15 and the marker concentration, which is usually a linear relationship, the thrombus marker concentration in the sample to be tested can be determined by calculating the resonant frequency offset.

[0055] Specifically, the thrombosis marker concentration detection includes the following steps:

[0056] Calibration curve acquisition:

[0057] (1) Build the above-mentioned coagulation function and thrombosis marker combined detection platform and prepare a thrombosis marker calibration sample with a concentration gradient range;

[0058] (2) Prepare or prepare a magnetoelastic sensor modified with a specific antibody, place the magnetoelastic sensor in the sample slot 142 of the mounting base 14, and insert the mounting base 14 into the detection coil 12. The steps for preparing or preparing a magnetoelastic sensor modified with a specific antibody are as follows: Figure 5Specifically, the prepared or obtained magnetoelastic sensor is first cleaned and dried with ethanol or deionized water to remove organic matter or impurities attached to the surface. A gold film is deposited on the clean magnetoelastic sensor using a process such as magnetron sputtering or electron beam thermal evaporation, with a thickness controlled to 60±10 nm (range 10-100 nm). Cysteine is covalently bonded to the gold film via the thiol-gold chemical self-assembly principle, introducing an amino group at its terminus. Streptavidin is then introduced via a condensation reaction of carboxyl and amino groups catalyzed by EDC and NHS. The amino group of the amino-polyethylene glycol-biotin reacts with the carboxyl group of the antibody to form an amide bond, achieving biotinylation of the antibody. Finally, the antibody is immobilized on the sensor surface through the specific binding of streptavidin and biotin. The above steps result in a magnetoelastic sensor for capturing specific thrombosis marker molecules (antigens). In this embodiment, the tetravalent binding properties of streptavidin (each molecule contains four biotin binding sites) are utilized to achieve high-density immobilization of antibodies, which can help improve the detection sensitivity of the sensor. In this embodiment, different specific antibodies can be modified by the above method to achieve the detection of different thrombosis marker molecules, including but not limited to D-dimer, thrombin-antithrombin complex (TAT), plasmin-α2 plasmin inhibitor complex (PIC), tissue plasminogen activator (t-PA), thrombomodulin (TM), and fibrinogen degradation products (FDP);

[0059] (3) The static magnetic field generating structure 11 generates a static magnetic field for the magnetoelastic sensor 15 to operate. The detection coil 12 generates an alternating magnetic field around the magnetoelastic sensor 15. The magnetic lines of force of the static magnetic field are parallel to the magnetic lines of force of the alternating magnetic field. The magnetoelastic sensor 15 resonates in the alternating magnetic field. The impedance analysis device 30 collects the electrical characteristics of the detection coil 12 and records the initial resonant frequency of the magnetoelastic sensor 15.

[0060] (4) Add 30-50 μl of calibration sample into the groove where the magnetoelastic sensor is located, insert it into the detection coil 12, and wait for 15-20 minutes;

[0061] (5) Figure 6 As shown, the specific antibodies modified on the surface of the magnetoelastic sensor 15 capture thrombus marker molecules in the sample being tested, causing the load mass on the sensor surface to change, thereby changing the resonant frequency of the sensor. The impedance analysis device 30 collects the electrical characteristics of the detection coil 12, records the response resonant frequency of the magnetoelastic sensor 15, and calculates the offset of the resonant frequency.

[0062] (6) Clean the magnetoelastic sensor 15 and place it in the sample slot at its base.

[0063] (7) Repeat steps 4-6;

[0064] (8) According to the above steps, the resonance frequency offset corresponding to different thrombus marker concentrations is obtained, and the calibration curve is calculated, as shown in the attached figure. Figure 7 As shown by the dotted line in .

[0065] Calculation of sample thrombosis marker concentration:

[0066] (1) Establishing the above-mentioned combined detection platform for coagulation function and thrombosis markers;

[0067] (2) Prepare or fabricate a magnetoelastic sensor modified with a specific antibody, place the magnetoelastic sensor in the sample slot 142 of the mounting base 14 , and insert the mounting base 14 into the detection coil 12 .

[0068] (3) The static magnetic field generating structure 11 generates a static magnetic field for the magnetoelastic sensor 15 to operate. The detection coil 12 generates an alternating magnetic field around the magnetoelastic sensor 15. The magnetic lines of force of the static magnetic field are parallel to the magnetic lines of force of the alternating magnetic field. The magnetoelastic sensor 15 resonates in the alternating magnetic field. The impedance analysis device 30 collects the electrical characteristics of the detection coil 12 and records the initial resonant frequency of the magnetoelastic sensor 15.

[0069] (4) Add 30-50 ml of the whole blood sample to be tested into the groove where the magnetoelastic sensor is located, and insert it into the detection coil 12, and wait for 15-20 minutes;

[0070] (5) Figure 6 As shown, the specific antibodies modified on the surface of the magnetoelastic sensor 15 capture thrombus marker molecules in the sample being tested, causing the load mass on the sensor surface to change, thereby changing the resonant frequency of the sensor. The impedance analysis device 30 collects the electrical characteristics of the detection coil 12, records the response resonant frequency of the magnetoelastic sensor 15, and calculates the offset of the resonant frequency.

[0071] (6) According to the calibration curve of the sensor's resonant frequency offset and the thrombus marker concentration, the thrombus marker concentration in the sample is calculated. The concentration of a thrombus marker in the sample is as follows: Figure 7 As shown in the asterisk mark.

[0072] The principle of coagulation function test is as follows Figure 8As shown, when blood coagulates under the action of an in vitro activator, its viscoelastic properties continuously change. When a magnetoelastic sensor operates in continuously coagulating blood, the energy generated by the magnetoelastic sensor in its resonant state is lost due to the viscous damping effect, causing changes in the sensor's electrical properties, namely, a decrease in its impedance amplitude. Therefore, a thromboelastogram is generated by monitoring the real-time impedance amplitude curve of the magnetoelastic sensor 15 to characterize the blood coagulation process. After data processing, coagulation-related parameters can be extracted from the time-impedance curve.

[0073] Specifically, the coagulation function test includes the following steps:

[0074] (1) Establishing the above-mentioned combined detection platform for coagulation function and thrombosis markers;

[0075] (2) preparing or fabricating the magnetoelastic sensor 15 and placing it in the sample slot 142 of the mounting base 14;

[0076] (3) Prepare anticoagulated whole blood or collect fresh human blood, and mix the whole blood sample with the activator in proportion;

[0077] (4) Immediately after the preparation is completed, take 30-50 μl of the mixed sample and add it to the groove where the magnetoelastic sensor 15 is located, and insert it into the detection coil 12;

[0078] (5) The static magnetic field generating structure 11 generates a static magnetic field for the magnetoelastic sensor 15 to operate. The detection coil 12 generates an alternating magnetic field around the magnetoelastic sensor 15. The magnetic lines of force of the static magnetic field are parallel to the magnetic lines of force of the alternating magnetic field. The magnetoelastic sensor 15 resonates in the alternating magnetic field. When the blood coagulates under the action of the activator, the viscoelasticity of the blood will continue to change. When the magnetoelastic sensor 15 operates in the continuously coagulating blood, due to the viscous damping effect, the energy generated by the magnetoelastic sensor 15 in the resonant state will be lost, and the impedance amplitude will decrease. The impedance analysis device 30 collects the electrical characteristics of the detection coil 12 and records the impedance amplitude of the magnetoelastic sensor 15.

[0079] (6) After collecting data for 15 minutes, or after the processor determines whether the stop condition is met, data collection is stopped and the magnetoelastic sensor 15 is removed;

[0080] (7) After the processor processes the data, it obtains the time-impedance amplitude change curve and extracts the coagulation-related parameters. According to the above steps, the time-impedance amplitude change curve of the blood coagulation process and the coagulation-related parameters are obtained. Figure 9 As shown, the coagulation-related parameters are defined as follows:

[0081] The reaction time ME-R is the time corresponding to the minimum of the second-order derivative of the time-impedance change curve;

[0082] The coagulation time ME-K is the difference between the time corresponding to the minimum of the first derivative of the time-impedance change curve and the coagulation time ME-R;

[0083] The clot formation rate ME-α is the angle between the zero baseline of the time-impedance change curve and the line connecting the coagulation rate ME-k;

[0084] The clot strength ME-A is the average change in impedance amplitude near the end point of the time-impedance change curve.

[0085] The combined detection platform for coagulation function and thrombus markers of the present application can obtain the concentration of thrombus markers through the frequency change of the magnetoelastic sensor 15 based on the mass load effect, and can obtain the thromboelastogram through the impedance amplitude change of the magnetoelastic sensor 15 based on the viscous damping effect, thereby analyzing and evaluating the coagulation disease status. Compared with existing detection methods, it has the advantages of being more efficient, more comprehensive and more accurate.

[0086] The above embodiments merely illustrate several implementations of the present invention, and while their descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the patented invention. It should be noted that those skilled in the art would be able to make numerous variations and improvements without departing from the spirit of the present invention. These variations and improvements are equivalent modifications and improvements to the above embodiments based on the essential technology of the present invention and fall within the scope of protection of the present invention.

Claims

1. A combined coagulation function and thrombosis marker detection platform, characterized by: The device comprises a detection device, a temperature control device and an impedance analysis device. The detection device comprises a static magnetic field generating structure, a detection coil, a heating component and a magnetoelastic sensor. The heating component is used to heat the blood sample. The magnetoelastic sensor is located at the center of the detection coil. The detection coil is electrically connected to the impedance analysis device. A current is applied to the detection coil to generate an alternating magnetic field for the operation of the magnetoelastic sensor. The static magnetic field generating structure is used to generate a static magnetic field for the operation of the magnetoelastic sensor. The magnetic lines of force of the static magnetic field are parallel to the magnetic lines of force of the alternating magnetic field. The impedance analysis device drives the detection coil to generate the alternating magnetic field around the magnetoelastic sensor. The magnetoelastic sensor resonates in the alternating magnetic field. The detection coil generates an induction signal, and the impedance analysis device collects the induction signal.

2. The combined coagulation function and thrombosis marker detection platform according to claim 1, characterized in that: The static magnetic field generating structure is a permanent magnet or a Helmholtz coil.

3. The combined coagulation function and thrombosis marker detection platform according to claim 1, characterized in that: When the detection device performs thrombus marker detection, the surface of the magnetoelastic sensor is modified with specific antibodies.

4. The combined coagulation function and thrombosis marker detection platform according to claim 1, characterized in that: When the detection device performs a coagulation function test, the magnetoelastic sensor is located in the blood that is in the coagulation process.

5. The combined coagulation function and thrombosis marker detection platform according to claim 1, characterized in that: The heating assembly includes a heat-conducting column and a heating belt wound around the heat-conducting column, and the magnetoelastic sensor is located in the heat-conducting column.

6. The combined coagulation function and thrombosis marker detection platform according to claim 5, characterized in that: The detection device further comprises a mounting base, which is detachably mounted inside the heat-conducting column and is used to carry the magnetoelastic sensor and the blood sample.

7. The combined coagulation function and thrombosis marker detection platform according to claim 5, characterized in that: The heating component is located inside the detection coil.

8. The combined coagulation function and thrombosis marker detection platform according to claim 5, characterized in that: The heating belt is a copper belt wrapped with polyimide.

9. The combined coagulation function and thrombosis marker detection platform according to claim 1, characterized in that: There are two detection devices, each of which is connected to the temperature control device and the impedance analysis device respectively. The two detection devices respectively perform coagulation function detection and thrombus marker detection on the blood sample.

10. The combined coagulation function and thrombosis marker detection platform according to claim 1, characterized in that: The number of the detection device is one, and the detection device performs coagulation function detection and thrombus marker detection on the blood sample at different times.

11. A detection method based on the coagulation function and thrombosis marker combined detection platform according to any one of claims 1 to 10, characterized in that: The following steps are involved: Thrombus marker concentration detection: A magnetoelastic sensor modified with a specific antibody is prepared and placed inside a detection coil of a detection device. A static magnetic field generating structure generates a static magnetic field in which the magnetoelastic sensor operates. The detection coil generates an alternating magnetic field around the magnetoelastic sensor. The magnetic lines of force of the static magnetic field are parallel to the magnetic lines of force of the alternating magnetic field. The magnetoelastic sensor resonates in the alternating magnetic field. The impedance analysis device collects the electrical characteristics of the detection coil and records the initial resonant frequency of the magnetoelastic sensor. A blood sample is added, so that the magnetoelastic sensor modified with specific antibodies is located in the blood sample. The specific antibodies modified on the surface of the magnetoelastic sensor capture thrombus marker molecules in the sample being tested, causing the load mass on the sensor surface to change, thereby changing the resonant frequency of the sensor. The impedance analysis device collects the electrical characteristics of the detection coil and records the response resonant frequency of the magnetoelastic sensor. Calculating the offset of the resonance frequency, obtaining a calibration curve based on the offset of the resonance frequency corresponding to different thrombosis marker concentrations, and calculating the thrombosis marker concentration contained in the tested sample; Coagulation function test: A blood sample and an activator are prepared in proportion, and the mixed sample is added so that the magnetoelastic sensor is located in the mixed sample, and the mixed sample is placed inside the detection coil of the detection device; the static magnetic field generating structure generates a static magnetic field for the operation of the magnetoelastic sensor, and the detection coil generates an alternating magnetic field around the magnetoelastic sensor. The magnetic lines of force of the static magnetic field are parallel to the magnetic lines of force of the alternating magnetic field, and the magnetoelastic sensor resonates in the alternating magnetic field. When the blood coagulates under the action of the activator, the viscoelasticity of the blood will continue to change. When the magnetoelastic sensor works in the continuously coagulating blood, due to the viscous damping effect, the energy generated by the magnetoelastic sensor in the resonant state will be lost, and the impedance amplitude will decrease. The impedance analysis device collects the electrical characteristics of the detection coil, records the impedance amplitude of the magnetoelastic sensor, obtains a time-impedance amplitude change curve, and extracts coagulation-related parameters.

12. The method for combined detection of coagulation function and thrombosis markers according to claim 11, characterized in that: The thrombus marker concentration detection step and the coagulation function detection step are respectively performed on the two detection devices.

13. The method for combined detection of coagulation function and thrombosis markers according to claim 11, characterized in that: The thrombus marker concentration detection step and the coagulation function detection step are performed on a detection device.

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