Multi-item coagulation test strip and matching detection device

Abstract

Disclosed in the present invention are a multi-item coagulation test strip and a matching detection device. The multi-item coagulation test strip comprises a main body, wherein first accommodating grooves are arranged on the main body, the number of the first accommodating grooves is equal to that of detection items of the test strip, and lyophilized beads matched to the detection items are correspondingly placed in the first accommodating grooves; the main body further comprises a plurality of second accommodating grooves, and functional reagents that react with the lyophilized beads are respectively accommodated in the second accommodating grooves; and a sealing film for sealing the first accommodating grooves and the second accommodating grooves is further provided on the surface of the main body. The functional reagents and the lyophilized beads are separately placed to avoid mutual interference, and agents mainly reacting with blood samples are lyophilized beads in a solid form, which significantly enhances the storage capacity and can ensure enzyme activity. The sealing film for sealing the first accommodating grooves and the second accommodating grooves is further provided on the surface of the main body, such that the test strip, after being packaged by means of the sealing film, can be stored and transported at room temperature, thereby improving the convenience of transportation.

Description

A coagulation multi-item joint detection test strip and an adaptive detection device TECHNICAL FIELD

[0001] The present application relates to the field of coagulation reagent strips, in particular to a coagulation multi-item joint detection test strip and an adaptive detection device. BACKGROUND

[0002] In the field of medical detection and research, in the process of detecting blood, after obtaining a blood sample (such as earlobe or fingertip peripheral blood, venous blood) from a subject, in order to avoid coagulation of the blood sample during storage or transfer, the blood is usually subjected to anticoagulation treatment, such as the addition of sodium citrate and other anticoagulants (blood agents).

[0003] Blood coagulation screening tests are required in clinical detection and routine screening, including detection of prothrombin time (PT), activated partial thromboplastin time (APTT), activated clotting time (ACT), fibrinogen content (FIB), and thrombin time (TT). Various detection instruments have been provided in the art to test the type of blood coagulation characteristics under specific conditions by clinical laboratories, health practitioners, etc.; in the prior art, a detection reagent corresponding to the utility model patent with application number 201711147040.9 is disclosed, which can be used for chemiluminescence immunoassay and solid-phase enzyme immunoassay. The detection reagents required for detection are all loaded on the detection reagent strip, realizing the integration of the reagent strip, simplifying the detection steps, avoiding contamination, and enabling effective qualitative or quantitative detection analysis of multiple index substances; however, in actual use, it is found that the reagents matched with the reagent strip are all in liquid form, and the activity of enzymes / proteins is difficult to maintain during detection, thus causing a large error in the measurement result. Therefore, a more reasonable solution is needed to solve the above technical problems. TECHNICAL PROBLEM

[0004] In view of the technical problem that in the test strip of the prior art, since the reagents matched are all in liquid form, the activity of enzymes / proteins is difficult to maintain during detection, thus causing a large error in the measurement result, the present application provides a solution. TECHNICAL SOLUTION

[0005] The application provides a blood coagulation multi-item joint test strip, which comprises a main body, a first accommodating groove is arranged on the main body, and the profile forming the first accommodating groove is transparent material, the number of the first accommodating grooves is equal to the detection items of the test strip, and a freeze-dried bead suitable for detecting an item is placed in each first accommodating groove; the main body further comprises a second accommodating groove, the second accommodating groove is provided with a plurality of second accommodating grooves, and each second accommodating groove contains a functional reagent reacting with the freeze-dried bead; and the surface of the main body is further provided with a sealing film for sealing the first accommodating groove and the second accommodating groove.

[0006] As an improved scheme of the application, the freeze-dried bead is obtained by the following preparation method:

[0007] Configuration: according to the type of the detection item to obtain corresponding components, and reconstitute to form a liquid reagent;

[0008] Droplet preparation: the liquid reagent is formed into a liquid droplet with a desired volume, and the liquid droplet is rapidly frozen by liquid nitrogen to obtain an initial freeze-dried bead;

[0009] Freeze-drying: the freeze-dried bead is collected for freeze-drying treatment, and after the freeze-drying treatment is completed, the freeze-dried bead with a loose reticular structure is formed.

[0010] As an improved scheme of the application, in the freeze-drying step, the parameters of the freeze-drying treatment are:

[0011] Pre-freezing stage: temperature: -45℃-40℃; time length: 3-6h;

[0012] Sublimation stage: temperature: -30℃-35℃; time length: 6-11h; vacuum degree <10pa;

[0013] Desorption drying: temperature: 5℃-25℃; time length: 4-10h; vacuum degree <10pa.

[0014] As an improved scheme of the application, in the freeze-drying step, the sublimation stage is divided into:

[0015] First sublimation stage: temperature: -30℃-35℃; time length: 4-7h; vacuum degree <10pa;

[0016] Second sublimation stage: temperature: -20℃-25℃; time length: 2-4h; vacuum degree <10pa.

[0017] As an improved scheme of the application, in the freeze-drying step, the desorption drying is divided into:

[0018] First desorption stage: temperature: 5℃-10℃; time length: 2-4h; vacuum degree <10pa;

[0019] The second analysis stage: temperature: 20-25℃; time: 2-6h; vacuum degree: <10pa.

[0020] As an improved scheme of the present application, the type of the detection item is APTT, PT, TT and FIB.

[0021] As an improved scheme of the present application, the freeze-dried bead for detecting the APTT item is a silica freeze-dried bead, which is compounded by buffer, silica, brain phospholipid, stabilizer, preservative and plasticizer.

[0022] The freeze-dried bead for detecting the PT item is a tissue factor freeze-dried reagent, which is compounded by buffer, recombinant tissue factor rhTF, stabilizer, preservative and plasticizer.

[0023] The freeze-dried bead for detecting the TT item is a bovine thrombin freeze-dried reagent, which is compounded by buffer, silica, brain phospholipid, stabilizer, preservative and plasticizer.

[0024] The freeze-dried bead for detecting the FIB item is a standard bovine thrombin freeze-dried reagent, which is compounded by buffer, bovine thrombin, stabilizer, preservative and plasticizer.

[0025] As an improved scheme of the present application, the functional reagent is a reconstitution solution, APTT reagent R1 and sample diluent.

[0026] As an improved scheme of the present application, the inner wall width of the first accommodating groove gradually increases from the groove bottom to the groove opening.

[0027] The present application also provides a detection device for detecting any of the above-mentioned coagulation multi-item joint detection test strips. Advantages

[0028] Compared with the prior art, the coagulation multi-item joint test strip and the matched detection device provided by the application have the following advantages: the coagulation multi-item joint test strip comprises a main body, the main body is provided with first accommodating grooves, the profiles forming the first accommodating grooves are transparent materials, the number of the first accommodating grooves is equal to the detection items of the test strip, and each first accommodating groove is correspondingly provided with a freeze-dried bead matched with a detection item; the main body is further provided with second accommodating grooves, the second accommodating grooves are provided with a plurality of functional reagents reacting with the freeze-dried beads; the surface of the main body is further provided with a sealing film used for sealing the first accommodating grooves and the second accommodating grooves; the functional reagents and the freeze-dried beads are separately placed to prevent mutual interference, and the reagent mainly reacting with the blood sample is a solid freeze-dried bead, which significantly enhances the storage capacity and can ensure the activity of the enzyme; further, the surface of the main body is further provided with a sealing film used for sealing the first accommodating grooves and the second accommodating grooves, and the sealing film can be used for normal-temperature storage and transportation after encapsulation, thereby increasing the transportation convenience. BRIEF DESCRIPTION OF DRAWINGS

[0029] Fig. 1 is a perspective view of the application;

[0030] Fig. 2 is another perspective view of the application;

[0031] Fig. 3 is a schematic view of the detection device of the application.

[0032] The main element symbols are explained as follows:

[0033] 1, first accommodating groove; 2, second accommodating groove; 3, Tip groove 3; 4, sample groove; 5, detection device;

[0034] 51, access assembly; 52, puncture needle; 53, sample injection needle; 54, light-emitting assembly. Best mode of the application

[0035] In the prior art, since the types of reagents matched are all in liquid form, the activity of enzymes / proteins is difficult to maintain during detection, thereby causing a technical problem of a large error in measurement results. In order to solve the above technical problem, the application provides a coagulation multi-item joint detection test strip. Referring to FIGS. 1 to 3, the joint detection test strip comprises a main body, the main body is provided with a first containing groove 1, and the profile forming the first containing groove 1 is of a transparent material. The number of the first containing grooves 1 is equal to the detection items of the test strip, and each first containing groove 1 is correspondingly provided with a freeze-dried bead suitable for the detection item. The main body further comprises a second containing groove 2 provided with a plurality of second containing grooves 2, and each second containing groove 2 contains a functional reagent reacting with the freeze-dried bead. The surface of the main body is further provided with a sealing film for sealing the first containing groove 1 and the second containing groove 2. The first containing groove 1 is used for storing the freeze-dried bead which mainly reacts with the blood sample, and also serves as a detection site. Therefore, the profile of this part is particularly transparent. During detection, a specific frequency band of light is used for irradiation, so as to correspondingly detect the required value. Naturally, in a better implementation, a Tip head hole can be further provided on the main body for placing a pipette, and a sample groove can be further provided for collecting a blood sample. The sealing film seals the Tip head hole and the sample groove, so as to ensure the cleanliness of the hole wall. In a specific use process, a technician first tears the sealing film of the sample groove and the Tip groove part, collects the blood sample of a person to be detected in the sample groove, and installs the pipette into the sample needle of a detection device 5. During work, the joint reagent strip is first placed on an in-out assembly 51 of the detection device 5. The in-out assembly 51 moves the joint reagent strip to a detection position based on a driving motor. During detection, a puncture needle 52 of the detection device 5 punctures the sealing film of the first containing groove 1. The blood sample and the functional reagent are added to the first containing groove 1 through the sample needle. The blood sample reacts with the freeze-dried bead, and is irradiated by a light emitting assembly 54 of the detection device 5. A signal receiver of the detection device 5 collects the light receiving condition of the blood sample to generate a corresponding light signal. A computing chip of the detection device 5 analyzes the coagulation degree and time of the blood sample according to the light signal, and thus completes the detection of the blood sample.

[0036] The functional reagent and the freeze-dried bead are separately placed to prevent mutual interference. The main reagent reacting with the blood sample is the freeze-dried bead in a solid form, which significantly enhances the storage capacity and can ensure the activity of the enzyme. Further, the surface of the main body is further provided with a sealing film for sealing the first containing groove 1 and the second containing groove 2. After being packaged by the sealing film, it can be stored and transported at room temperature, thereby increasing the convenience of transportation. Embodiment of the application

[0037] In a specific implementation, the freeze-dried bead is obtained by the following preparation method:

[0038] S1, configuration: according to the type of detection project to obtain the corresponding components, and compound to form a liquid reagent;

[0039] S2, making droplets: let the liquid reagent form a liquid drop of expected volume, and quickly freeze the liquid drop by liquid nitrogen to obtain an initial frozen bead;

[0040] S3, freeze-drying: collect the freeze-dried beads and perform freeze-drying treatment, and after the freeze-drying treatment, form a freeze-dried bead with a loose network structure.

[0041] The preparation of the freeze-dried bead is realized through the above steps. Since the material containing a large amount of water is frozen into a solid by pre-cooling in the freeze-drying stage, and then the solid water is directly sublimated under vacuum conditions, the volume of the material remains unchanged after drying, forming a loose porous network structure. According to the degree of freeze-drying, the longer the freeze-drying time, the higher the degree of loose freeze-dried beads, and the higher the dissolution rate in use. Based on the freeze-drying technology, the enzyme activity is not affected, and after mixing with functional additives, the re-dissolution is rapid, and a stable packaging environment can be formed for the measurement of blood samples.

[0042] More specifically, in the S1 configuration step, the type of detection project is APTT, PT, TT and FIB; the freeze-dried bead for detecting the APTT project is a silica method freeze-dried bead, which is compounded by buffer, silica, brain phospholipid, stabilizer, preservative and plasticizer; in terms of components, the buffer is 0.02-1M, the silica mass accounts for 0.01-1w / v% of the buffer volume; the brain phospholipid mass accounts for 0.01-1w / v% of the buffer volume; the stabilizer mass accounts for 0.1-5w / v% of the buffer volume; the preservative mass accounts for 0.03-1w / v% of the buffer volume; and the excipient mass accounts for 0.01-1w / v% of the buffer volume.

[0043] The freeze-dried bead for detecting the PT project is a tissue factor freeze-dried reagent, which is compounded by buffer, recombinant tissue factor rhTF, stabilizer, preservative and plasticizer; wherein, in terms of components, the buffer is 0.02-1M; the recombinant tissue factor mass accounts for 0.1-1w / v% of the buffer volume; the stabilizer mass accounts for 0.1-5w / v% of the buffer volume; the preservative mass accounts for 0.03-1w / v% of the buffer volume; and the excipient mass accounts for 0.01-1w / v% of the buffer volume.

[0044] The lyophilized beads for detecting TT item are bovine thrombin lyophilized reagent, which is compounded by buffer, silica, brain phospholipid, stabilizer, preservative and plasticizer; wherein, in terms of components, the buffer is 0.02-1M; the bovine thrombin is 10-60U / mL; the stabilizer accounts for 0.1-5w / v% of the volume of the buffer; the preservative accounts for 0.03-1w / v% of the volume of the buffer; and the excipient accounts for 0.01-1w / v% of the volume of the buffer.

[0045] The lyophilized beads for detecting FIB item are standard bovine thrombin lyophilized reagent, which is compounded by buffer, bovine thrombin, stabilizer, preservative and plasticizer; wherein, in terms of components, the buffer is 0.02-1M; the bovine thrombin is 18-30U / mL; the stabilizer accounts for 0.1-5w / v% of the volume of the buffer; the preservative accounts for 0.03-1w / v% of the volume of the buffer; and the excipient accounts for 0.01-1w / v% of the volume of the buffer.

[0046] The preservative accounts for 0.03-1w / v% of the volume of the buffer; and the excipient accounts for 0.01-1w / v% of the volume of the buffer.

[0047] More specifically, in the S3 step, the lyophilized beads compounded by the above components can quickly detect blood samples and can simultaneously detect the above four coagulation items, which is more efficient and fast.

[0048] More specifically, in the S2 preparation of droplets, the prepared liquid reagent is divided into 2-30ul droplets by a micro pump, which can be quickly frozen into a frozen bead state by liquid nitrogen at this volume;

[0049] The lyophilized bead manufacturing instrument is operated as follows:

[0050] 1) Start the machine to start the droplet quantification system and log in the system;

[0051] 2) Connect the droplet quantification system with the special droplet pipeline and needle for each item;

[0052] 3) Rinse the droplet pipe and needle;

[0053] 4) After the rinsing is completed, start the air compressor, set the droplet parameters, adjust the droplet target volume, set the parameters, and prepare the droplet;

[0054] 5) Perform droplet by a micro pump to obtain initial frozen beads.

[0055] More specifically, in the S3 lyophilization step, the parameters for lyophilization treatment are as follows:

[0056] Pre-freezing stage: temperature: -45℃-40℃; time: 3-6h;

[0057] Sublimation stage: temperature: -30℃-35℃; duration: 6-11h; vacuum degree <10pa;

[0058] Desorption stage: temperature: 5℃-25℃; duration: 4-10h; vacuum degree <10pa.

[0059] By the above pre-freezing stage, sublimation stage or desorption stage, the water molecule substance is sublimated in large quantities, and the core reaction substance is retained for use, and the reaction substance is in a loose and porous state, which is convenient for reaction with the functional reagent.

[0060] In the preferred scheme, in the freeze-drying step, the sublimation stage is divided into:

[0061] First sublimation stage: temperature: -30℃-35℃; duration: 4-7h; vacuum degree <10pa;

[0062] Second sublimation stage: temperature: -20℃-25℃; duration: 2-4h; vacuum degree <10pa.

[0063] In the preferred scheme, in the freeze-drying step, the desorption stage is divided into:

[0064] First desorption stage: temperature: 5℃-10℃; duration: 2-4h; vacuum degree <10pa;

[0065] Second desorption stage: temperature: 20℃-25℃; duration: 2-6h; vacuum degree <10pa.

[0066] By dividing the sublimation stage and the desorption stage into two stages, a total of four small stages are used to process the initial frozen beads, so that the overall freeze-drying degree is more reasonable, and the strength of the loose and porous structure of the freeze-dried beads is better.

[0067] In this embodiment, the functional reagent is a cleaning solution, a reconstitution solution, an APTT reagent R1 and a sample diluent. Specifically, the sample diluent is composed of imidazole buffer and preservative, wherein the imidazole buffer is 0.05-3M; the mass of the preservative accounts for 0.03-1w / v% of the volume of the imidazole buffer; the reconstitution solution is composed of buffer and sodium hypochlorite, wherein the buffer is 0.02-1M, and the mass of sodium chloride accounts for 0.03-0.1w / v% of the volume of the solution; the cleaning solution is composed of sodium hypochlorite and surfactant, wherein the volume of sodium hypochlorite accounts for 5-30v / v% of the volume of the solution, and the mass of the surfactant accounts for 0.01-1w / v% of the volume of the solution.

[0068] The above-mentioned component types are:

[0069] The reconstitution solution can be HEPES, PBS buffer, Tris buffer, physiological saline, purified water or distilled water.

[0070] The sample diluent includes but is not limited to imidazole, barbital, sodium chloride, preservative, etc.

[0071] The cleaning solution includes but is not limited to sodium hypochlorite, stabilizer, etc.

[0072] The above stabilizer includes but is not limited to sucrose, fructose, BSA, trehalose, amino acid, etc.

[0073] The above preservative includes but is not limited to PC300, DN, KV100, KV300, carboxymethylcellulose, etc.

[0074] The above plasticizer includes but is not limited to mannitol, dextran, PEG8000, PEG6000, etc.

[0075] The above buffer solution pH is 6-9, including but not limited to Tris buffer, HEPES buffer, PBS buffer, etc.

[0076] The above surfactant includes but is not limited to Tween series - Tween 20, 80, 60, etc., Triton series, Triton 15, 100, 405, etc.

[0077] In the embodiment, the inner wall width of the first accommodating groove 1 gradually increases from the groove bottom to the groove opening. It is not difficult to understand that, in the detection, the freeze-dried beads are placed at the bottom of the first accommodating groove 1. Since the first accommodating groove 1 has a certain taper, the freeze-dried beads can be immersed in a smaller volume of functional reagents and blood samples to react, and a large operation space can be obtained, thereby increasing the working stability of the puncture needle and the sampling needle 53.

[0078] The application also provides a detection device 5 for detecting any coagulation multi-item joint test strip. The detection device 5 includes an inlet and outlet group, a puncture needle 52, a sampling needle, a light-emitting assembly 54 and a signal collector. For details, reference can be made to the foregoing content, which will not be described here.

[0079] The advantages of the application are:

[0080] 1. The functional reagents and freeze-dried beads are placed separately to prevent mutual interference, and the main reagent reacting with the blood sample is a solid freeze-dried bead, which significantly enhances the storage capacity and can ensure the activity of the enzyme. Further, the surface of the main body is also provided with a sealing film for sealing the first accommodating groove and the second accommodating groove. After being packaged by the sealing film, it can be stored and transported at room temperature, thereby increasing the transportation convenience.

[0081] 2. Because the freeze-drying stage involves pre-freezing substances containing a large amount of water into solids, and then allowing the solid water to sublimate directly under vacuum conditions, while the substance itself remains in the ice frame from which it was frozen, its volume remains unchanged after drying, forming a loose and porous network structure. This does not damage the activity of proteins / enzymes, while allowing for rapid reconstitution and making measurement more convenient.

[0082] The above-disclosed embodiments are merely a few specific examples of the present invention, but the present invention is not limited thereto. Any variations that can be conceived by those skilled in the art should fall within the protection scope of the present invention.

Claims

1. A coagulation multi-item simultaneous test strip, characterized by, The application relates to a multi-item coagulation test strip, which comprises a main body, a first accommodating groove arranged on the main body, and a transparent material forming a profile of the first accommodating groove, wherein the number of the first accommodating grooves is equal to the detection items of the test strip, and each of the first accommodating grooves is used for placing a freeze-dried bead suitable for the detection item; the main body further comprises a second accommodating groove, a plurality of the second accommodating grooves are arranged, and each of the second accommodating grooves is used for accommodating a functional reagent reacting with the freeze-dried bead; and the surface of the main body is further provided with a sealing film used for sealing the first accommodating groove and the second accommodating groove.

2. The coagulation multi-item simultaneous testing strip according to claim 1, wherein, The freeze-dried bead is obtained by the following preparation method. Configuration: according to the type of the detection item, corresponding components are obtained, and a liquid reagent is formed by compounding; Droplet preparation: the liquid reagent is made into a liquid droplet with a desired volume, and the liquid droplet is rapidly frozen by liquid nitrogen to obtain an initial freeze-dried bead; Freeze-drying: the freeze-dried bead is collected and subjected to freeze-drying treatment, and after the freeze-drying treatment is completed, the freeze-dried bead with a loose net structure is formed.

3. The coagulation multi-item simultaneous test strip according to claim 2, characterized in that, In the freeze-drying step, the parameters of the freeze-drying treatment are as follows: Pre-freezing stage: temperature: -45 DEG C to -40 DEG C; time length: 3-6h; Sublimation stage: temperature: -30 DEG C to -35 DEG C; time length: 6-11h; vacuum degree: <10pa; Desorption drying: temperature: 5 DEG C to 25 DEG C; time length: 4-10h; vacuum degree: <10pa. In the freeze-drying step, the sublimation stage is divided into: First sublimation stage: temperature: -30 DEG C to -35 DEG C; time length: 4-7h; vacuum degree: <10pa; Second sublimation stage: temperature: -20 DEG C to -25 DEG C; time length: 2-4h; vacuum degree: <10pa.

4. The coagulation multi-item simultaneous testing strip according to claim 2, wherein, In the freeze-drying step, the desorption drying is divided into: First desorption stage: temperature: 5 DEG C to 10 DEG C; time length: 2-4h; vacuum degree: <10pa; Second desorption stage: temperature: 20 DEG C to 25 DEG C; time length: 2-6h; vacuum degree: <10pa. The type of the detection item is APTT, PT, TT and FIB. The freeze-dried bead for detecting the APTT item is a silica freeze-dried bead, which is compounded by a buffer, silica, brain phospholipid, a stabilizer, a preservative and a plasticizer; 5. The coagulation multi-item joint test strip according to claim 2, characterized in that, The freeze-dried bead for detecting the PT item is a tissue factor freeze-dried reagent, which is compounded by a buffer, a recombinant tissue factor rhTF, a stabilizer, a preservative and a plasticizer; The freeze-dried bead for detecting the TT item is a bovine thrombin freeze-dried reagent, which is compounded by a buffer, silica, brain phospholipid, a stabilizer, a preservative and a plasticizer; The freeze-dried bead for detecting the FIB item is a standard bovine thrombin freeze-dried reagent, which is compounded by a buffer, bovine thrombin, a stabilizer, a preservative and a plasticizer. The functional reagent is a reconstitution solution, APTT reagent R1 and a sample diluent. The inner wall width of the first accommodating groove gradually increases from the groove bottom to the groove opening.

6. The coagulation multi-item simultaneous test strip according to claim 1, wherein, The application is used for detecting the coagulation multi-item coagulation test strip.

7. The coagulation multi-item simultaneous test strip according to claim 2, wherein, ​ ​ ​ ​ 8. The coagulation multi-item simultaneous test strip according to claim 1, wherein, ​ 9. The coagulation multi-item simultaneous test strip according to claim 1, wherein, ​ 10. A detection device, characterized in that ​

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