Platelet adhesion function detection system and detection method

Abstract

The invention relates to the technical field of platelet adhesion functions, in particular to a platelet adhesion function detection system and method. The platelet adhesion function detection system comprises a detection module, the detection module comprises a detection unit, the detection unit comprises a processor, at least one analysis detection chamber and at least one standard detection chamber, in the detection process, a blood sample in the analysis detection chamber is converted into an activated state from a non-activated state, and the activated state is converted into an activated state from a non-activated state. And the processor outputs a platelet adhesion function detection result according to the change of the optical signal characteristics of the analysis detection chamber and the standard detection chamber. According to the present invention, the microfluidic technology and the turbidimetry technology are combined to achieve the automatic detection of the platelet adhesion aggregation function, such that the influence of the fibrinogen in the blood on the platelet adhesion function detection is avoided, and the accuracy of the platelet adhesion function detection is improved.

Description

Technical Field

[0001] The present invention relates to the technical field of platelet adhesion function, and in particular to a platelet adhesion function detection system and detection method. Background Art

[0002] Platelet adhesion function is an indicator for evaluating the adhesion ability of platelets in the blood. Platelet adhesion usually occurs after blood vessels are damaged. Platelets stay in the damaged blood vessels through surface receptors and the interaction with adhesion molecules such as collagen or other exposed matrix proteins under the endothelium. Platelet adhesion function is the first step for platelets to reach the site of injury and initiate hemostasis. Therefore, the detection of platelet adhesion function is of great significance for diagnosing hemorrhagic diseases and monitoring the effect of antiplatelet therapy. At present, most of them use glass bead method or fibrin binding test to simulate the adhesion process of platelets at the site of vascular injury and quantify their functional status. However, the operation steps are complicated and there are serious errors in the test results.

[0003] The prior art discloses a method of utilizing the ability of platelets to form thrombi under specific conditions and evaluating the platelet adhesion function by measuring the time that the thrombus blocks the micropores. However, since fibrinogen in the blood is converted into fibrin during the coagulation process, the formation of platelet thrombi is promoted, resulting in a shortened measured closure time. At the same time, red blood cells in the blood not only cause changes in blood viscosity, but also affect the distribution and aggregation efficiency of platelets on damaged blood vessel walls. Therefore, the presence of fibrinogen and red blood cells in the blood affects the closure time, resulting in inaccurate detection of platelet adhesion function. Summary of the invention

[0004] Therefore, the technical problem to be solved by the present invention is to overcome the defect of inaccurate platelet adhesion function detection results in the prior art, thereby providing a platelet adhesion function detection system and detection method.

[0005] To this end, on the one hand, the present invention provides a platelet adhesion function detection system, including a detection module, the detection module includes a detection unit, the detection unit includes a processor, at least one analysis detection chamber and at least one standard detection chamber. During the detection process, the blood sample in the analysis detection chamber is changed from an inactive state to an active state, and the processor outputs the platelet adhesion function detection result according to the change of the optical signal characteristics of the analysis detection chamber and the standard detection chamber.

[0006] In some of the embodiments, the analysis and detection chamber includes a stirring mechanism, an optical detection mechanism, an optical signal conversion mechanism and a receiver.

[0007] In some of the embodiments, the standard detection chamber includes an optical detection mechanism, an optical signal conversion mechanism, and a receiver.

[0008] In some of the embodiments, the optical detection mechanism includes a light source and an optical detection channel provided with an adsorption material. Preferably, the adsorption material is fibrinogen and von Willebrand factor.

[0009] In some of the embodiments, the optical signal conversion mechanism includes optical particles coated with biomaterials, the biomaterials are capable of binding to platelet surface receptors, and the biomaterials include polypeptides, proteins and their derivatives.

[0010] In some embodiments, the wavelength of the light source is greater than or equal to 700 nm; the optical characteristic peak of the optical particles is greater than or equal to 700 nm, and the size of the optical particles is 1 nm-100 μm.

[0011] In some embodiments, based on the volume of each detection chamber, the number of optical particles in the analysis detection chamber and / or the standard detection chamber is 5×10 5 / mL-5×10 10 Pieces / mL.

[0012] In some embodiments, the stirring mechanism includes a stirrer and a driver for controlling a stirring rate of the stirrer.

[0013] In some of the embodiments, the detection module further comprises a control detection unit, wherein the control detection unit comprises at least one blank detection chamber, wherein the blank detection chamber comprises a light source, a receiver and an optical detection channel.

[0014] In some of the embodiments, the number of the stirring bar is at least one, and the ratio of the volume of the stirring bar to the volume of the blood sample in the analysis and detection chamber is 1:(10-1000).

[0015] In some embodiments, the detection system further comprises a sample processing module, wherein the sample processing module is used to perform preliminary processing on the blood sample.

[0016] Preferably, the sample processing module includes at least one of a sample tube storage area or a sample heating area.

[0017] Preferably, the stirrer is at least one of a stainless steel ball, a ferrite, and a neodymium iron boron magnet.

[0018] Preferably, the driver is a magnet.

[0019] On the other hand, the present invention provides a method for detecting platelet adhesion function, in which a blood sample is injected into the above-mentioned platelet adhesion function detection system, and the processor outputs the platelet adhesion function detection result according to the changes in the optical signal characteristics obtained by the receivers in the analysis detection chamber and the standard detection chamber.

[0020] In some of these embodiments, the blood sample is tested in the system for 0.5 min to 10 min;

[0021] In some of the embodiments, the receiver in the analysis and detection chamber acquires the optical signal characteristics of the blood sample in real time under stirring, and the stirring rate is 100rpm-5000rpm.

[0022] Preferably, the optical signal characteristic includes at least one of the optical signal, the slope of the optical signal change curve, and the area of ​​the optical signal change curve.

[0023] In some embodiments, the platelet adhesion function is evaluated by platelet reaction time.

[0024] Preferably, the platelet reaction time is the time when the maximum change of the light signal is 50-100%.

[0025] In some of the embodiments, before the blood sample is injected into the detection module, a step of heating the blood sample in a sample heating area is included, and the heating temperature is 36-40°C.

[0026] In some of the embodiments, the optical signal characteristic is a parameter after removing the background signal obtained by the control detection unit.

[0027] The technical solution of the present invention has the following advantages:

[0028] 1. A platelet adhesion function detection system provided by the present invention comprises a detection module, wherein the detection module comprises a detection unit, wherein the detection unit comprises a processor, at least one analysis detection chamber and at least one standard detection chamber. During the detection process, the blood sample in the analysis detection chamber changes from an inactive state to an active state, and the processor outputs the platelet adhesion function detection result according to the change of the optical signal characteristics of the analysis detection chamber and the standard detection chamber. The present invention realizes the automatic detection of platelet adhesion and aggregation function by combining microfluidics technology with turbidimetry technology, thereby avoiding the influence of fibrinogen in the blood on the platelet adhesion function detection and improving the accuracy of the platelet adhesion function detection.

[0029] 2. The present invention provides a platelet adhesion function detection system, wherein the optical detection mechanism includes a light source and an optical detection channel provided with an adsorption material, wherein the adsorption material is fibrinogen and von Willebrand factor; the optical signal conversion mechanism includes optical particles, wherein the optical particles are coated with biomaterials, wherein the biomaterials are capable of binding to platelet surface receptors, and wherein the biomaterials include polypeptides, proteins, and their derivatives. The present invention utilizes optical particles coated with biomaterials to specifically adhere to free platelets on the one hand, and to adhere to platelets adhered to the optical detection channel on the other hand, thereby affecting the change of the optical signal characteristics of the detection channel. At the same time, a standard detection chamber is used as a deduction for the unactivated platelet adhesion function to obtain a detection result that is not affected by fibrinogen in the blood.

[0030] 3. A platelet adhesion function detection method provided by the present invention, wherein a blood sample is injected into the platelet adhesion function detection system, and the processor outputs the platelet adhesion function detection result according to the change of the optical signal characteristics obtained by the receivers in the analysis detection chamber and the standard detection chamber. The detection method provided by the present invention can avoid the problem of abnormal platelet closure time caused by fibrinogen and red blood cells in the blood, thereby obtaining accurate platelet adhesion function detection results. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0032] Figure 1 is a schematic diagram of a platelet adhesion detection system provided in a first embodiment of the present invention;

[0033] Figure 2 is a schematic diagram of a platelet adhesion detection system provided in a second embodiment of the present invention;

[0034] Figure 3 It is a schematic diagram of the structure of the analysis and detection chamber provided in the third embodiment of the present invention;

[0035] Figure 4 is a schematic diagram of the structure of a standard detection chamber provided in a fourth embodiment of the present invention;

[0036] Figure 5 is a schematic diagram of the structure of a blank detection chamber provided in a fifth embodiment of the present invention;

[0037] Figure 6is a schematic diagram of a platelet adhesion detection system in Example 2 of the present invention;

[0038] Figure 7 is a diagram of a light signal fitting curve pattern in Example 2 of the present invention;

[0039] Figure 8 is a schematic diagram of a platelet adhesion detection system in Example 3 of the present invention;

[0040] Fig. 9 is a linear fitting graph of the abnormal coagulation blood sample results in the experimental example of the present invention;

[0041] Fig.10 It is a linear fitting diagram of the results of normal blood coagulation samples in the experimental example of the present invention;

[0042] Reference numerals:

[0043] 11-first light source; 12-second light source; 13-third light source;

[0044] 21-first optical detection channel; 22-second optical detection channel; 23-third optical detection channel;

[0045] 31-first receiver; 32-second receiver; 33-third receiver;

[0046] 41-stirring bar; 61-driver;

[0047] 51-first optical signal conversion mechanism; 52-second optical signal conversion mechanism;

[0048] 100-sample tube storage area; 10-puncture needle; 200-sample heating area;

[0049] 300-standard testing room; 400-analysis testing room; 401-first analysis testing room;

[0050] 402-second analysis and detection room; 500-blank detection room. DETAILED DESCRIPTION

[0051] The following examples are provided for a better understanding of the present invention, but are not intended to limit the best mode of implementation, nor to limit the content and protection scope of the present invention. Any product identical or similar to the present invention obtained by anyone under the inspiration of the present invention or by combining the features of the present invention with other prior arts shall fall within the protection scope of the present invention.

[0052] If no specific experimental steps or conditions are specified in the examples, the conventional experimental steps or conditions described in the literature in the field can be used. If no manufacturer is specified for the reagents or instruments used, they are all conventional reagent products that can be obtained commercially.

[0053] See also Figure 1 As shown, the present invention provides a platelet adhesion function detection system, including a detection module, wherein the detection module includes a detection unit, and the detection unit includes a processor, at least one analysis detection chamber and at least one standard detection chamber.

[0054] The processor outputs the platelet adhesion function test result according to the change of the optical signal characteristics between the analysis test chamber and the standard test chamber.

[0055] The change in the optical signal characteristic includes at least one of a change amount of the optical signal, a slope of an optical signal change curve, and an area of ​​an optical signal change curve.

[0056] The platelet adhesion function test result is the platelet closure time, and the platelet closure time is the reaction time when the change amount of the light signal is 50-100%.

[0057] The analysis and detection chamber comprises a stirring mechanism, an optical detection mechanism, an optical signal conversion mechanism and a receiver.

[0058] The standard detection chamber comprises an optical detection mechanism, an optical signal conversion mechanism and a receiver, and the standard detection chamber is used to detect the platelet adhesion function in an unactivated cell state.

[0059] Specifically, the stirring mechanism includes a stirrer and a driver for controlling the stirring rate of the stirrer.

[0060] The stirrer is at least one of a stainless steel ball, a ferrite, and a neodymium iron boron magnet. The stirring mechanism includes a stirrer. The number of stirrers in the detection chamber is at least 1. Exemplarily, the number of stirrers is 1, 2, 3, 4, 5, or 6.

[0061] The present invention does not limit the shape of the stirrer, which may be a sphere, a cone, or a rod, preferably a sphere.

[0062] The ratio of the volume of the stirring bar to the volume of the blood sample in the analysis and detection chamber is 1:(10-1000), preferably 1:(20-100), and more preferably 1:50.

[0063] The driver for controlling the stirring speed of the stirring bar is a magnet, which is arranged at the bottom of the detection chamber and is used to drive the stirring bar to shear the blood sample in the detection chamber to activate platelets.

[0064] The optical detection mechanism comprises a light source and an optical detection channel provided with an adsorption material, wherein the adsorption material is at least one of fibrinogen and von Willebrand factor.

[0065] The wavelength of the light source is greater than or equal to 700nm. Using a light source with a wavelength greater than or equal to 700nm can avoid the influence of red blood cells in the blood on the characteristic parameters of the light signal. Exemplarily, the wavelength of the light source is 700nm, 800nm, 850nm, 900nm, and 1000nm.

[0066] An optical detection channel is arranged on the inner wall of the detection chamber, and an adsorption material is loaded on the optical detection channel, and the adsorption material can adhere to platelets.

[0067] The present invention does not limit the position and number of the optical detection channels, as long as they can adhere to platelets.

[0068] The optical detection channel is arranged on the opposite side of the light source, and can also be arranged on both sides of the light source.

[0069] The method of loading the adsorption material on the optical detection channel is at least one of physical adsorption and chemical cross-linking.

[0070] Specifically, the physical adsorption method is to use the volume of each detection chamber as 100 mL / m 3 (protein solution volume: each detection chamber volume) is used, and a protein solution with a concentration of 1 mg / mL is coated on the optical detection channel, dried, and repeated three times to achieve loading the adsorption material on the optical detection channel. For example, when the volume of the detection chamber is 100 μL, 0.01 mL of 1 mg / mL protein solution is coated on the area of ​​the optical detection channel in the detection chamber, dried, and the above steps are repeated three times to load the optical detection channel with the protein adsorption material.

[0071] Specifically, the chemical cross-linking method comprises the following steps: loading carboxyl groups on the optical detection channel after surface plasma treatment, and calculating the volume of each detection chamber at 100 mL / m 3 (Volume of each solvent: volume of each detection chamber) By adding 1 volume of 10 mg / mL 1-ethyl-(3-dimethylaminopropyl)carbodiimide (EDC) and 1 volume of 10 mg / mL N-hydroxysuccinimide (NHS) and 0.1 volume of 20 mg / mL protein solution, the interaction between protein and amino group is achieved, and the adsorbent material is loaded on the optical detection channel. For example, when the volume of the detection chamber is 100 μL, 1 volume is 0.01 mL.

[0072] Furthermore, the specific operation method of surface plasma treatment can refer to the manual of the surface plasma cleaning machine. For example, the plasma cleaning machine nozzle is aimed at the surface that needs to be activated in the optical detection channel of each detection chamber, and high-purity carbon dioxide is used as the working gas. The spraying is continued for 1-5 minutes to load the surface with carboxyl groups.

[0073] Specifically, the optical signal conversion mechanism includes optical particles.

[0074] Based on the volume of each detection chamber, the number of optical particles in the analysis detection chamber and / or the standard detection chamber is 5×10 5 / mL-5×10 10 Pieces / mL.

[0075] The optical characteristic peak of the optical particles is greater than or equal to 700 nm. Exemplarily, the optical characteristic peak of the optical particles is 700 nm, 800 nm, 900 nm and 1000 nm.

[0076] The size of the optical particles is 1 nm-100 μm, preferably 20-60 μm, and more preferably 50 μm.

[0077] The optical particles are coated with biological materials, which can bind to platelet surface receptors. The biological materials include polypeptides, proteins and derivatives thereof.

[0078] Specifically, the protein is at least one of fibrinogen, fibronectin, vitronectin, collagen, laminin, and von Willebrand factor (vWF), and the polypeptide is a fibrinogen polypeptide, a fibronectin polypeptide, a vitronectin polypeptide, a collagen polypeptide, a laminin polypeptide, and von Willebrand factor (vWF) polypeptide.

[0079] The derivatives are derivatives of the above-mentioned proteins or polypeptides.

[0080] The method of coating the biomaterial with the optical particles comprises at least one of a chemical covalent cross-linking method and a physical adsorption method.

[0081] Among them, the chemical covalent cross-linking method is to dilute the optical particles into a 1 mg / mL suspension with 10mM pH4.0 PBS, take 1 volume of 10mg / mL EDC solution (dissolved in 10mM pH4.0 PBS) and 10mg / mL NHS solution (dissolved in 10mM pH4.0 PBS), vortex mix, and stir and activate for 30min at room temperature. Centrifuge and remove the supernatant. Then dissolve the binding peptide of the platelet surface glycoprotein receptor into a 10mg / mL solution with 10mM pH7.4 PBS, redisperse the optical particles with 1 volume of 10mM pH7.4 PBS, add 1 volume of binding peptide solution, vortex mix, and stir for 4h at room temperature. Centrifuge and remove the supernatant. Add 2 volumes of 10mg / mL bovine serum albumin solution and stir and seal at room temperature for 30min. Centrifuge and remove the supernatant to obtain optical particles coated with binding peptides of platelet surface glycoprotein receptors. Dilute to a suitable concentration with a suitable buffer for detection.

[0082] The physical adsorption method is to dilute the optical particles into a 1 mg / mL suspension with 10 mM PBS at pH 7.4, and then dissolve the binding peptide of the platelet surface glycoprotein receptor into a 10 mg / mL solution with 10 mM PBS at pH 7.4. Take 1 volume of the optical particle suspension and 1 volume of the binding peptide solution, vortex to mix, and stir overnight at room temperature. Centrifuge to remove the supernatant. Add 2 volumes of 10 mg / mL bovine serum albumin solution and stir at room temperature for 30 minutes. Centrifuge to remove the supernatant to obtain the optical particles coated with the binding peptide of the platelet surface glycoprotein receptor. Dilute with a suitable buffer to a suitable concentration for detection.

[0083] Specifically, the volume of each reagent in the method of coating a biological material with optical particles is related to the capacity of each detection chamber.

[0084] The analysis and detection chamber also includes a receiver for receiving optical signals. The present invention does not limit the position of the receiver as long as it can receive the optical signal of the optical detection channel.

[0085] See also Figure 2 As shown, the detection module also includes a control detection unit, which includes at least one blank detection chamber. The blank detection chamber includes a light source, a receiver and an optical detection channel. The blank detection chamber is used to detect background signals and remove errors of background signals.

[0086] The volumes of the standard detection chamber, the analytical detection chamber and the blank detection chamber are 80-200 μL.

[0087] The platelet adhesion function detection system also includes a sample processing module, which includes a sample tube storage area. The sample tube storage area is used to perform preliminary processing on the blood sample. A puncture needle is provided in the sample tube storage area. The puncture needle is used to puncture the blood sample and transport the blood sample to the detection module. The sample tube storage area is provided to facilitate the storage of blood samples, so that the blood samples can be taken out at any time for use.

[0088] Furthermore, the sample processing module also includes a sample heating area, which heats the blood sample to 36-40°C and delivers the heated blood sample to the detection module. By setting the sample heating area, the temperature of the blood sample can be adjusted to the same temperature as when the sample was taken, avoiding the test results of the platelet adhesion function being affected by too low or too high temperature.

[0089] Specifically, the present invention provides an analysis and detection chamber, the specific structure of which is shown in Figure 3 The analysis and detection chamber includes a first light source 11 and a first optical detection channel 21 arranged on the inner wall of the analysis and detection chamber. The first optical detection channel 21 is loaded with an adsorption material, and the adsorption material can adhere to platelets.

[0090] The present invention does not limit the position and number of the first optical detection channel, as long as it can achieve platelet adhesion and reflect the changes in the characteristics of the optical signal in the presence of a light source.

[0091] Specifically, the first optical detection channel is arranged on the opposite side of the first light source, and may also be arranged on both sides of the first light source.

[0092] The method of loading the adsorption material on the first optical detection channel is at least one of physical adsorption and chemical cross-linking.

[0093] Specifically, the physical adsorption method is to use the volume of the analysis and detection chamber as 100mL / m 3 The amount of protein solution with a concentration of 1 mg / mL is coated on the optical detection channel, dried, and repeated three times to achieve loading the adsorption material on the optical detection channel. For example, when the volume of the analysis detection chamber is 100 μL, 0.01 mL of 1 mg / mL protein solution is coated on the area of ​​the optical detection channel in the detection chamber, dried, and the above steps are repeated three times to load the optical detection channel with the protein adsorption material.

[0094] Specifically, the chemical cross-linking method comprises the following steps: loading carboxyl groups on the optical detection channel after surface plasma treatment at a rate of 100 mL / m 3 The adsorbent material was loaded on the optical detection channel by adding 1 volume of 10 mg / mL 1-ethyl-(3-dimethylaminopropyl)carbodiimide (EDC), 1 volume of 10 mg / mL N-hydroxysuccinimide (NHS) and 0.1 volume of 20 mg / mL protein solution to achieve the interaction between protein and carboxyl group.

[0095] The analysis and detection chamber further comprises a first receiver 31 for receiving optical signals. The present invention does not limit the position of the first receiver, as long as it can receive the optical signal of the first optical detection channel.

[0096] The analysis and detection chamber is provided with a first optical signal conversion mechanism 51. Specifically, the first optical signal conversion mechanism is an optical particle. The surface of the optical particle is coated with a biological material. The biological material can bind to the platelet surface receptor. The biological material includes polypeptides, proteins and their derivatives.

[0097] The analysis and detection chamber also includes a stirring mechanism. The present invention does not limit the shape, size, etc. of the stirring mechanism, as long as it can shear the blood sample to activate platelets.

[0098] The stirring mechanism includes a first stirrer 41 and a first driver 61 for controlling the stirring rate of the first stirrer. The driver is arranged at the bottom of the detection chamber and is used to drive the first stirrer to shear the blood sample in the analysis detection chamber to activate platelets. The driver can be a magnet.

[0099] Specifically, the present invention provides a standard detection chamber, the specific structure of which is shown in Figure 4 The standard detection chamber includes a second light source 12 and a second optical detection channel 22 arranged on the inner wall of the analysis and detection chamber. The second optical detection channel 22 is loaded with an adsorption material, and the adsorption material can adhere to platelets.

[0100] Specifically, the second optical detection channel is arranged on the opposite side of the second light source, and may also be arranged on both sides of the second light source.

[0101] The standard detection chamber further includes a second receiver 32 for receiving optical signals. The present invention does not limit the position of the second receiver as long as it can receive the optical signal of the second optical detection channel.

[0102] The standard detection chamber is provided with a second optical signal conversion mechanism 52. Specifically, the second optical signal conversion mechanism also includes optical particles. The surface of the optical particles is coated with biological materials. The biological materials can bind to platelet surface receptors. The biological materials include polypeptides, proteins and their derivatives.

[0103] Specifically, the present invention provides a blank detection chamber, the specific structure of which is shown in Figure 5 The blank detection chamber includes a third light source 13 and a third optical detection channel 23 arranged on the inner wall of the analysis detection chamber. The third optical detection channel 23 is loaded with an adsorption material, and the adsorption material can adhere to platelets.

[0104] Specifically, the third optical detection channel is arranged on the opposite side of the third light source, and may also be arranged on both sides of the third light source.

[0105] The blank detection chamber further includes a third receiver 33 for receiving optical signals. The present invention does not limit the position of the third receiver as long as it can receive the optical signal characteristics of the third optical detection channel.

[0106] The present invention provides a method for detecting platelet adhesion function, comprising the following steps:

[0107] Blood samples are injected into the analysis detection chamber and the standard detection chamber in the detection module respectively, and the processor outputs the platelet adhesion function test according to the changes in the optical signal characteristics obtained from the receivers in the analysis detection chamber and the standard detection chamber.

[0108] The detection time of blood samples in the detection module is 0.5min-10min;

[0109] To activate platelets, in some embodiments, the receiver in the analysis and detection chamber acquires the optical signal of the blood sample in real time under stirring, and the stirring rate is 100rpm-5000rpm.

[0110] The change in the optical signal characteristic includes at least one of a change amount of the optical signal, a slope of an optical signal change curve, and an area of ​​an optical signal change curve.

[0111] The platelet adhesion function is evaluated by platelet reaction time.

[0112] The platelet reaction time is the time when the maximum change of the light signal reaches 50-100%.

[0113] Before the blood sample is injected into the detection module, the method further includes heating the blood sample in a sample heating area, wherein the heating temperature is 36-40°C.

[0114] Example 1

[0115] See also Figure 2 As shown, this embodiment provides a platelet adhesion function detection system, including a sample processing module and a detection module, wherein the sample processing module includes a sample tube storage area and a sample heating area.

[0116] The detection module comprises a standard detection unit, an analysis detection unit and a control detection unit, wherein the blood sample in the analysis detection unit is stirred during the detection so that the platelets are in an activated state.

[0117] The standard detection unit is provided with an optical detection mechanism, an optical signal conversion mechanism and a receiver;

[0118] The analysis and detection unit is provided with a stirring mechanism, an optical detection mechanism, an optical signal conversion mechanism and a receiver;

[0119] The comparison detection unit is provided with an optical detection mechanism and a receiver.

[0120] During use, the blood sample is pierced through the rubber in the sample tube cap by the puncture needle in the sample tube storage area, so that the blood sample is injected into the sample heating area. After the blood sample is heated to 37°C, the blood sample is injected into the standard detection unit, the analysis detection unit and the control detection unit respectively. The receiver in the standard detection unit obtains the characteristic parameters of the light signal of the blood sample with unactivated platelets, the receiver in the analysis detection unit obtains the characteristic parameters of the light signal of the blood sample with activated platelets, and the receiver in the control detection unit obtains the background signal.

[0121] The processor fits the optical signals obtained by each receiver into an optical signal variation curve, and records the time taken when the area of ​​the optical signal variation curve reaches 90% as the platelet reaction time.

[0122] Example 2

[0123] See also Figure 6 As shown, this embodiment provides a platelet adhesion function detection system, including a sample processing module and a detection module, wherein the sample processing module includes a sample tube storage area 100 and a sample heating area 200, wherein the sample tube storage area is provided with a puncture needle 10.

[0124] The detection module includes a standard detection unit, an analysis detection unit and a control detection unit, wherein the standard detection unit includes a standard detection chamber 300, the analysis detection unit is provided with two analysis detection chambers, namely a first analysis detection chamber 401 and a second analysis detection chamber 402, and the control detection unit includes a blank detection chamber.

[0125] The first analysis and detection chamber and the second analysis and detection chamber have the same structure. Figure 3 As shown, each of the analysis and detection chambers includes a first light source 11, a first optical detection channel 21 disposed on the inner wall of the analysis and detection chamber, the first optical detection channel 21 is loaded with an adsorption material, the adsorption material can adhere to platelets, the first optical detection channel is disposed on the opposite side of the first light source, and the analysis and detection chamber also includes a first receiver 31 for receiving optical signals. The analysis and detection chamber also includes optical particles, the surface of the optical particles is coated with a biological material, the biological material is fibrinogen, the size of the optical particles is 50 μm, and the number is 5×10 5 / mL (based on the volume of the analysis and detection chamber). The analysis and detection chamber also includes a stirring mechanism, which includes a first stirring bar 41 and a first driver 61 for controlling the stirring rate of the first stirring bar, wherein the first stirring bar is a stainless steel ball, and the driver is arranged at the bottom of the detection chamber, and the driver is a magnet.

[0126] The structure of the standard test chamber is shown in Figure 4 As shown, the standard detection chamber includes a second light source 12 and a second optical detection channel 22 arranged on the inner wall of the analysis and detection chamber. The second optical detection channel 22 is loaded with an adsorption material, and the adsorption material can adhere to platelets. The adsorption material is fibrinogen, and the light source is an LED lamp with a wavelength of 800nm. The second optical detection channel is arranged on the opposite side of the second light source. The standard detection chamber also includes a second receiver 32 for receiving optical signals. The standard detection chamber also includes optical particles, the surface of the optical particles is coated with biological materials, and the biological material is fibrinogen. The size of the optical particles is 50μm and the number is 5×105 / mL (based on the volume of the standard detection chamber).

[0127] The structure of the blank detection chamber is shown in Figure 5 As shown, it includes a third light source 13 and a third optical detection channel 23 arranged on the inner wall of the analysis and detection chamber, the light source is an LED lamp with a wavelength of 800nm, and the third optical detection channel 23 is loaded with an adsorption material, the adsorption material can adhere to platelets, and the adsorption material is fibrinogen. The third optical detection channel is arranged on the opposite side of the third light source, and the blank detection chamber also includes a third receiver 33 for receiving optical signals.

[0128] In this embodiment, the volumes of the analysis detection chamber, the standard detection chamber, and the blank detection chamber are all 100 μL.

[0129] The method for loading the adsorbent material in the optical detection channel of the standard detection chamber, the analytical detection chamber and the blank detection chamber is as follows: based on the volume of the standard detection chamber, the analytical detection chamber and the blank detection chamber, the adsorbent material is loaded at 100 mL / m 3 The amount (protein solution volume: each detection chamber volume) was used to apply a protein solution with a concentration of 1 mg / mL on the optical detection channel of each detection chamber, and dried, and repeated three times.

[0130] The method for coating biomaterials with optical particles in the standard detection chamber and the analytical detection chamber is to dilute the optical particles into a 1 mg / mL suspension with 10 mM PBS at pH 7.4, and then dissolve fibrinogen into a 10 mg / mL binding peptide solution with 10 mM PBS at pH 7.4. Take 1 volume of the optical particle suspension and 1 volume of the binding peptide solution, vortex to mix, and stir overnight at room temperature. Centrifuge to remove the supernatant. Add 2 volumes of 10 mg / mL bovine serum albumin solution and stir at room temperature for 30 minutes. Centrifuge to remove the supernatant to obtain the optical particles coated with fibrinogen (the volume of each reagent is based on the volume of each detection chamber, according to 100 mL / m 3 Dosage, mL / m 3 is the volume of each reagent: detection chamber volume).

[0131] During the platelet adhesion function test, the blood sample is pierced through the rubber in the sample tube cap by the puncture needle in the sample tube storage area, and the blood sample is injected into the sample heating area. After the blood sample is heated to 37°C, the blood sample is injected into the standard test chamber, the first analysis test chamber, the second analysis test chamber and the blank test chamber respectively. After 5 minutes, the second receiver in the standard test chamber obtains the blood sample light signal of unactivated platelets, the first receiver in the second analysis test chamber and the third analysis test chamber obtains the blood sample light signal of activated platelets, and the third receiver in the blank test chamber obtains the background signal.

[0132] The processor fits the optical signals obtained by each receiver into an optical signal change curve, and records the time taken when the area of ​​the optical signal change curve is 90% as the platelet reaction time. Figure 7 Light signal fitting curve model, t 闭 is the platelet reaction time.

[0133] Example 3

[0134] See also Figure 8 This embodiment provides a platelet adhesion function detection system, including a sample processing module and a detection module, wherein the sample processing module includes a sample tube storage area 100 and a sample heating area 200, wherein the sample tube storage area is provided with a puncture needle 10.

[0135] The detection module includes a standard detection unit, an analysis detection unit and a control detection unit, wherein the standard detection unit includes a standard detection chamber 300 , the analysis detection unit includes an analysis detection chamber 400 , and the control detection unit includes a blank detection chamber 500 .

[0136] The structure of the analysis and detection chamber is shown in Figure 3 As shown, each of the analysis and detection chambers includes a first light source 11, a first optical detection channel 21 arranged on the inner wall of the analysis and detection chamber, the light source is an LED lamp with a wavelength of 800nm, the first optical detection channel 21 is loaded with an adsorption material, the adsorption material is von Willebrand factor, the first optical detection channel is arranged on the opposite side of the first light source, and the analysis and detection chamber also includes a first receiver 31 for receiving light signals. The analysis and detection chamber also includes optical particles, the surface of the optical particles is coated with biological materials, the biological materials are collagen, the size of the optical particles is 50μm, the optical characteristic peak of the optical particles is 800nm, and the number is 5×10 6 The analysis and detection chamber further comprises a stirring mechanism, which comprises a first stirring bar 41 and a first driver 61 for controlling the stirring rate of the first stirring bar, wherein the first stirring bar is a stainless steel ball, and the driver is disposed at the bottom of the detection chamber and is a magnet.

[0137] The structure of the standard test chamber is shown in Figure 4As shown, the standard detection chamber includes a second light source 12 and a second optical detection channel 22 arranged on the inner wall of the analysis and detection chamber. The second optical detection channel 22 is loaded with an adsorption material, and the adsorption material can adhere to platelets. The adsorption material is von Willebrand factor, and the light source is an LED lamp with a wavelength of 800nm. The second optical detection channel is arranged on the opposite side of the second light source. The standard detection chamber also includes a second receiver 32 for receiving optical signals. The standard detection chamber also includes optical particles, the surface of the optical particles is coated with biological materials, and the biological material is fibrinogen. The size of the optical particles is 50μm and the number is 5×10 6 / mL, and the optical characteristic peak of the optical particles is 800nm.

[0138] The structure of the blank detection chamber is shown in Figure 5 As shown, it includes a third light source 13 and a third optical detection channel 23 arranged on the inner wall of the analysis and detection chamber, the light source is an LED lamp with a wavelength of 800nm, and the third optical detection channel 23 is loaded with an adsorption material, the adsorption material can adhere to platelets, and the adsorption material is von Willebrand factor. The third optical detection channel is arranged on the opposite side of the third light source, and the blank detection chamber also includes a third receiver 33 for receiving optical signals.

[0139] In this embodiment, the volume of the analysis detection chamber, the standard detection chamber and the blank detection chamber is 150 μL.

[0140] Among them, the method for loading the adsorbent material on the optical detection channel in the standard detection chamber, the analytical detection chamber and the blank detection chamber is to load the carboxyl group on the optical detection channel after the optical detection channel is subjected to surface plasma, and to achieve the interaction between the protein and the amino group by adding 1 volume of 10 mg / mL 1-ethyl-(3-dimethylaminopropyl)carbodiimide (EDC) and 1 volume of 10 mg / mL N-hydroxysuccinimide (NHS) and 0.1 volume of 20 mg / mL protein solution, and load the adsorbent material on the optical detection channel (the volume of each reagent is calculated based on the volume of each detection chamber, according to 100 mL / m 3 Dosage, mL / m 3 is the volume of each reagent: detection chamber volume).

[0141] The method for coating biomaterials with optical particles in standard detection chambers and analytical detection chambers is to dilute the optical particles into a 1 mg / mL suspension with 10 mM PBS at pH 7.4, and then dissolve the collagen into a 10 mg / mL binding peptide solution with 10 mM PBS at pH 7.4. Take 1 volume of the optical particle suspension and 1 volume of the binding peptide solution, vortex to mix, and stir overnight at room temperature. Centrifuge to remove the supernatant. Add 2 volumes of 10 mg / mL bovine serum albumin solution and stir at room temperature for 30 minutes. Centrifuge to remove the supernatant to obtain optical particles coated with collagen (the volume of each reagent is based on the volume of each detection chamber, according to 100 mL / m 3 Dosage, mL / m 3 is the volume of each reagent: detection chamber volume).

[0142] During the platelet adhesion function test, the blood sample is pierced through the rubber in the sample tube cap by the puncture needle in the sample tube storage area, and the blood sample is injected into the sample heating area. After the blood sample is heated to 37°C, the blood sample is injected into the standard test chamber, the analysis test chamber and the blank test chamber respectively. After 10 minutes, the second receiver in the standard test chamber obtains the blood sample light signal of unactivated platelets, the first receiver in the analysis test chamber obtains the blood sample light signal of activated platelets, and the third receiver in the blank test chamber obtains the background signal.

[0143] The processor fits the optical signals obtained by each receiver into an optical signal variation curve, and records the time taken when the area of ​​the optical signal variation curve reaches 90% as the platelet reaction time.

[0144] Experimental example

[0145] Ten blood samples with abnormal coagulation function and ten blood samples with normal coagulation function were taken, and the platelet adhesion function detection system of Example 2 of the present invention was used as the experimental group, and the platelet function analysis system PFA200 was used as the control group for platelet adhesion function detection. The platelet closure time was used as the indicator for evaluating the platelet adhesion function in both the experimental group and the control group, and the closure time of 82s-150s was the normal closure time. The results are shown in Table 1.

[0146] Table 1 Platelet closure time determination

[0147]

[0148]

[0149] According to the data in Table 1, the data obtained from the experimental and control groups of blood samples with abnormal coagulation exceeded the standard threshold range of platelet closure time, and both showed abnormality. In the blood samples with normal coagulation, the control groups No. 1 and No. 7 showed abnormal coagulation, indicating that there was an error in the test results of the control group. Linear fitting was performed on the data obtained from the experimental and control groups of blood samples with abnormal coagulation function in Table 1, and we obtained Fig. 9 , Fig. 9 Medium linear correlation R 2 =0.9759, while the data of the blood sample experimental group and the control group (the control group excluded abnormal data) with normal coagulation function were linearly fitted to obtain Fig.10 , Fig.10 Medium linear correlation R 2 =0.9339, which proves that the detection system and method provided by the present invention have good detection and diagnosis effects and have certain application scenarios.

[0150] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the invention.

Claims

1. A platelet adhesion function detection system, characterized in that: The detection module comprises a detection unit, wherein the detection unit comprises a processor, at least one analysis detection chamber and at least one standard detection chamber. During the detection process, the blood sample in the analysis and detection chamber changes from an inactive state to an active state. The processor outputs the platelet adhesion function test result according to the change of the optical signal characteristics between the analysis test chamber and the standard test chamber.

2. The platelet adhesion function detection system according to claim 1, characterized in that: The analysis and detection chamber comprises a stirring mechanism, an optical detection mechanism, an optical signal conversion mechanism and a receiver; and / or, The standard detection chamber includes an optical detection mechanism, an optical signal conversion mechanism and a receiver.

3. The platelet adhesion function detection system according to claim 2, characterized in that: The optical detection mechanism comprises a light source and an optical detection channel provided with an adsorption material, wherein the adsorption material is at least one of fibrinogen and von Willebrand factor; and / or, The optical signal conversion mechanism comprises optical particles, wherein the optical particles are coated with biological materials, wherein the biological materials can bind to platelet surface receptors, and the biological materials comprise at least one of polypeptides, proteins and derivatives thereof.

4. The platelet adhesion function detection system according to claim 3, characterized in that: The wavelength of the light source is greater than or equal to 700 nm; and / or, The optical characteristic peak of the optical particles is greater than or equal to 700 nm, and the size of the optical particles is 1 nm-100 μm; and / or, Based on the volume of each detection chamber, the number of optical particles in the analysis detection chamber and / or the standard detection chamber is 5×10 5 / mL-5×10 10 / mL; and / or, The stirring mechanism comprises a stirring bar and a driver for controlling the stirring rate of the stirring bar.

5. The platelet adhesion function detection system according to claim 4, characterized in that: The detection module further comprises a control detection unit, wherein the control detection unit comprises at least one blank detection chamber, wherein the blank detection chamber comprises a light source, a receiver and an optical detection mechanism; and / or, The number of the stirring bars is at least one, and the ratio of the volume of the stirring bars to the volume of the blood sample in the analysis and detection chamber is 1:(10-1000).

6. The platelet adhesion function detection system according to claim 5, characterized in that: The detection system also includes a sample processing module, which is used to perform preliminary processing on the blood sample. The sample processing module includes at least one of a sample tube storage area or a sample heating area; The stirring bar is at least one of a stainless steel ball, a ferrite, and a neodymium iron boron magnet; The driver is a magnet.

7. A method for detecting platelet adhesion function, characterized in that: A blood sample is injected into the platelet adhesion function detection system according to any one of claims 1 to 6, and the processor outputs the platelet adhesion function detection result according to the change of the optical signal characteristics obtained by the receivers in the analysis detection chamber and the standard detection chamber.

8. The platelet adhesion function detection method according to claim 7, characterized in that: The blood sample detection time in the system is 0.5min-10min; and / or, Under the stirring state, the receiver in the analysis and detection chamber acquires the optical signal characteristics of the blood sample in real time, and the stirring rate is 100rpm-5000rpm.

9. The platelet adhesion function detection method according to claim 8, characterized in that: The optical signal feature includes at least one of the optical signal, the slope of the optical signal change curve, and the area of ​​the optical signal change curve; and / or, The platelet adhesion function is evaluated by platelet reaction time.

10. The platelet adhesion function detection method according to claim 9, characterized in that: The platelet reaction time is the time when the maximum change of the light signal is 50-100%; and / or, Before the blood sample is injected into the detection module, the method further includes heating the blood sample in a sample heating area, wherein the heating temperature is 36-40°C.

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