Multi-sample processing method for coagulation tester

Abstract

The invention discloses a multi-sample processing method for a coagulation tester, and the invention relates to the technical field of multi-sample processing. The specific operation steps of the multi-sample processing method for a coagulation tester are as follows: step one, collecting multiple coagulation samples and placing them in a specific sterile sample container. The advantages of the invention are: adding magnetic beads with specific ligands modified on the surface and incubating them, combined with stirring, can quickly and effectively remove impurities and interfering substances in plasma, using an ultrafiltration membrane with a specific pore size and appropriate pressure during the ultrafiltration process, can quickly remove large and small molecule interfering substances, and the chemical precipitation step can quickly precipitate trace impurities and remove them by centrifugation, and the subsequent adjustment of plasma concentration can also be completed quickly, so that the processed samples can be quickly put into the coagulation tester for detection. Compared with traditional processing methods, the method greatly shortens the sample detection time and improves the efficiency of the entire detection process.

Description

Technical Field

[0001] The invention relates to the technical field of multi-sample processing, in particular to a multi-sample processing method for a coagulation tester. Background Art

[0002] Coagulation tester is an important instrument widely used in the medical field to evaluate the coagulation function of human blood. Coagulation is a complex and critical physiological process involving the interaction of many coagulation factors to prevent excessive bleeding and maintain the integrity of blood vessels. The emergence of coagulation tester makes it possible to accurately evaluate this process. The working principle of coagulation tester is based on a variety of technologies. The most common ones are optical detection methods, which determine the coagulation time and related parameters by monitoring the changes in light scattering, transmission or absorption during blood coagulation. There are also electrochemical detection methods, which use electrochemical reactions on the electrode surface to detect material changes in the coagulation process. These testers are usually able to measure multiple coagulation-related indicators, such as prothrombin time (PT), partially activated thromboplastin time (APTT), fibrinogen content, etc. These indicators are of great significance for the diagnosis of various hemorrhagic and thrombotic diseases.

[0003] The existing multi-sample processing technology of coagulation testers has significant defects in time efficiency. When processing multiple samples, the entire process lacks systematic optimization, resulting in excessive time spent in each link, making multi-sample processing time-consuming, which not only wastes time but also increases the cost of testing. In addition, the existing coagulation testers perform poorly in the accuracy of multi-sample test results, affecting the accuracy of the test results. For this reason, we propose a multi-sample processing method for coagulation testers. Summary of the invention

[0004] The object of the present invention is to provide a multi-sample processing method for a coagulation tester.

[0005] In order to solve the problems raised in the above background technology, the present invention provides the following technical solution: a multi-sample processing method for a coagulation tester, wherein the specific operation steps of the multi-sample processing method for a coagulation tester are as follows:

[0006] Step 1: Collect multiple coagulation blood samples, place them in specific sterile sample containers, perform visual inspection on the samples, and remove samples with less than 2 ml of blood, coagulated samples (visible fibrin threads), and samples that are severely contaminated (visible foreign matter and color abnormalities).

[0007] Step 2: Transfer the sample after preliminary screening to a centrifuge device with temperature control function, and then start the centrifuge device to perform centrifugal operation to separate the sample into a plasma layer and a blood cell layer;

[0008] Step 3: Slowly draw out the separated plasma and transfer it to a new sterile container, add magnetic beads with surface modification that specifically recognize impurities and interfering substances to the plasma, and incubate at 23°C to 28°C for 30 to 35 minutes. Stir during the incubation process to adsorb the magnetic beads through an external magnetic field, thereby removing impurities and interfering substances in the plasma;

[0009] Step 4: The plasma treated with magnetic beads is passed through an ultrafiltration device, using an ultrafiltration membrane with a pore size of 0.22 μm and ultrafiltration at a pressure of 0.1 MPa to remove macromolecules with a molecular weight greater than 100 kDa and small molecule interferences with a molecular weight less than 1 kDa in the plasma;

[0010] Step 5: Add a 10% trichloroacetic acid solution to the ultrafiltered plasma, react for 18 to 22 minutes at a pH of 4.5 to 5.0 and a temperature of 37°C to 40°C to precipitate trace impurities, place in a centrifuge to remove the precipitate, and centrifuge at a speed of 3000 rpm to 3200 rpm for 8 to 12 minutes;

[0011] Step 6: Use a spectrophotometer to measure the concentration of the target substance in the treated plasma. According to the requirements of the coagulation test, when the concentration exceeds 5.0 mg / ml, it is diluted with 0.9% saline at a ratio of 1:1. When the concentration is lower than 0.5 mg / ml, it is concentrated by vacuum rotary evaporation to keep the plasma concentration between 0.5 mg / ml and 5.0 mg / ml. The treated sample is then placed in a coagulation tester for testing.

[0012] As a further solution of the present invention: In the step 2, the centrifugal equipment used has a temperature control function, and the centrifugal treatment is carried out at a rotation speed of 3000rpm to 3200rpm, the centrifugal time is 13min to 16min, and the centrifugal temperature is maintained at 4°C to 6°C. This low temperature can reduce the activity of enzymes in the blood and reduce the occurrence of biochemical reactions, thereby maintaining the original state of various components in the sample.

[0013] As a further solution of the present invention: In the step three, the magnetic beads are incubated with slight stirring at a speed of 90rpm to 110rpm, which can promote the uniform distribution of the magnetic beads in the plasma, avoid local aggregation of the magnetic beads, and increase the chance and efficiency of the magnetic beads binding to impurities.

[0014] As a further solution of the present invention: in the step 4, the flux of the ultrafiltration membrane is regularly monitored during the ultrafiltration process, and when the flux drops to 50% of the initial flux, the ultrafiltration membrane needs to be replaced immediately.

[0015] As a further solution of the present invention: in the step 5, after the chemical precipitation centrifugation reaction is completed, high performance liquid chromatography (HPLC) is used to detect the effect of precipitation removal to ensure that the plasma meets the quality requirements of the coagulation test after pretreatment.

[0016] As a further solution of the present invention: in step six, the diluted and concentrated plasma needs to be mixed again by vortex mixing for 2 minutes to 4 minutes to ensure that the concentration gradient is broken and the plasma is fully mixed.

[0017] As a further solution of the present invention: after each step from step 1 to step 6 is completed, the container and the operating tool are cleaned with a 75% ethanol solution to remove residual samples and processing reagents. The cleaning operation should follow strict operating procedures to ensure that the ethanol solution is in full contact with the container and the tool, and after cleaning, they should be thoroughly rinsed and dried to remove ethanol residues to avoid affecting subsequent sample processing.

[0018] As a further solution of the present invention: the containers and tools need to be dried at 60° C. to 80° C. for 25 min to 30 min after cleaning to ensure that the containers and tools are dry and sterile.

[0019] As a further solution of the present invention: in steps 1 to 6, the entire sample pretreatment process is carried out in an environment with a cleanliness level of 100, and the operators are required to wear disposable sterile protective clothing, gloves and masks that meet hygiene standards to ensure the sterility and safety of each operation.

[0020] By adopting the above technical solution, compared with the prior art, the beneficial effects of the present invention are:

[0021] 1. The present invention pre-treats the sample before the coagulation tester is tested, including a centrifugation step, and controls the temperature, rotation speed and time of the centrifugation device, so as to quickly and effectively separate the plasma layer and the blood cell layer, thereby saving time for subsequent processing. The magnetic beads with specific ligands on the surface are added and incubated, and stirred, so as to quickly and effectively remove impurities and interfering substances in the plasma. The ultrafiltration membrane with a specific pore size and appropriate pressure are used in the ultrafiltration process, so as to quickly remove macromolecular and small molecule interfering substances. The chemical precipitation step can quickly precipitate trace impurities and remove them by centrifugation. The subsequent adjustment of the plasma concentration can also be completed quickly, so that the treated sample can be quickly put into the coagulation tester for testing. Compared with the traditional processing method, the present method greatly shortens the time of sample testing and improves the efficiency of the entire testing process.

[0022] 2. The present invention excludes samples with insufficient blood volume, coagulation and serious contamination through strict appearance inspection standards. If these samples enter the detection process, they are very likely to cause deviations in the test results. The centrifugation operation is carried out under specific temperature conditions, which maintains the original state of various components in the sample and avoids biochemical reactions caused by improper temperature, thereby reducing the impact on the test results. The stirring during the magnetic bead incubation process and the reasonable incubation conditions ensure that the magnetic beads are fully combined with impurities and interfering substances, and these factors that affect the accuracy of the test are removed to the maximum extent. The ultrafiltration step can accurately remove interfering substances in a specific molecular weight range, further improving the purity of the plasma. The chemical precipitation step is carried out under strictly controlled pH, temperature and reaction time conditions, so that trace impurities can be effectively precipitated and removed, improving the quality of the plasma. In the concentration adjustment link after the treatment, the plasma concentration is accurately controlled within an appropriate range, ensuring the accuracy of the test results;

[0023] 3. The present invention eliminates samples that do not meet the requirements at the initial stage through strict sample screening steps, avoids subsequent useless processing operations on these samples, saves reagent and time costs, optimizes steps such as centrifugation, magnetic bead treatment, and ultrafiltration, thereby reducing the amount of reagents used and the operating time of the equipment while achieving the same processing effect. Through standardized cleaning and drying of containers and tools, their service life is extended and the replacement frequency is reduced, thereby saving procurement costs and generally reducing indirect costs caused by processing failures. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is a schematic diagram showing the characteristics comparison among Example 1, Example 2 and a control group in the embodiments of the present invention;

[0025] Figure 2 Schematic diagram of the method steps in an embodiment of the present invention. DETAILED DESCRIPTION

[0026] The specific embodiments of the present invention will be further described below in conjunction with the accompanying drawings. It should be noted that the description of these embodiments is used to help understand the present invention, but does not constitute a limitation of the present invention.

[0027] In addition, the technical features involved in the various embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0028] Please see attached Figure 1 -Attached Figure 2 The multi-sample processing method of the coagulation tester of the present invention, the specific operation steps of the multi-sample processing method of the coagulation tester are as follows:

[0029] Step 1: Collect multiple coagulation blood samples, place them in specific sterile sample containers, perform visual inspection on the samples, and remove samples with less than 2 ml of blood, coagulated samples (visible fibrin threads), and samples that are severely contaminated (visible foreign matter and color abnormalities).

[0030] Step 2: Transfer the sample after preliminary screening to a centrifuge device with temperature control function, and then start the centrifuge device to perform centrifugal operation to separate the sample into a plasma layer and a blood cell layer;

[0031] Step 3: Slowly draw out the separated plasma and transfer it to a new sterile container, add magnetic beads with surface modification that specifically recognize impurities and interfering substances to the plasma, and incubate at 23°C to 28°C for 30 to 35 minutes. Stir during the incubation process to adsorb the magnetic beads through an external magnetic field, thereby removing impurities and interfering substances in the plasma;

[0032] Step 4: The plasma treated with magnetic beads is passed through an ultrafiltration device, using an ultrafiltration membrane with a pore size of 0.22 μm and ultrafiltration at a pressure of 0.1 MPa to remove macromolecules with a molecular weight greater than 100 kDa and small molecule interferences with a molecular weight less than 1 kDa in the plasma;

[0033] Step 5: Add a 10% trichloroacetic acid solution to the ultrafiltered plasma, react for 18 to 22 minutes at a pH of 4.5 to 5.0 and a temperature of 37°C to 40°C to precipitate trace impurities, place in a centrifuge to remove the precipitate, and centrifuge at a speed of 3000 rpm to 3200 rpm for 8 to 12 minutes;

[0034] Step 6: Use a spectrophotometer to measure the concentration of the target substance in the treated plasma. According to the requirements of the coagulation test, when the concentration exceeds 5.0 mg / ml, it is diluted with 0.9% saline at a ratio of 1:1. When the concentration is lower than 0.5 mg / ml, it is concentrated by vacuum rotary evaporation to keep the plasma concentration between 0.5 mg / ml and 5.0 mg / ml. The treated sample is then placed in a coagulation tester for testing.

[0035] In one embodiment of the present invention: in step 2, the centrifugal equipment used has a temperature control function, and the centrifugal treatment is performed at a rotation speed of 3000rpm to 3200rpm, the centrifugal time is 13min to 16min, and the centrifugal temperature is maintained at 4°C to 6°C. This low temperature can reduce the activity of enzymes in the blood and reduce the occurrence of biochemical reactions, thereby maintaining the original state of various components in the sample.

[0036] In one embodiment of the present invention: In step three, the magnetic beads are incubated with slight stirring at a speed of 90 rpm to 110 rpm, which can promote the uniform distribution of the magnetic beads in the plasma, avoid local aggregation of the magnetic beads, and increase the chance and efficiency of the magnetic beads binding to impurities.

[0037] In one embodiment of the present invention: in step 4, the flux of the ultrafiltration membrane is regularly monitored during the ultrafiltration process, and when the flux drops to 50% of the initial flux, the ultrafiltration membrane needs to be replaced immediately.

[0038] In one embodiment of the present invention: in step five, after the chemical precipitation centrifugation reaction is completed, high performance liquid chromatography (HPLC) is used to detect the effect of precipitate removal to ensure that the plasma meets the quality requirements of the coagulation test after pretreatment.

[0039] In one embodiment of the present invention: in step six, the diluted and concentrated plasma needs to be mixed again by vortex mixing for 2 minutes to 4 minutes to ensure that the concentration gradient is broken and the plasma is fully mixed.

[0040] In one embodiment of the present invention: after each step from step 1 to step 6 is completed, the container and the operating tool are cleaned with a 75% ethanol solution to remove residual samples and processing reagents. The cleaning operation should follow strict operating procedures to ensure that the ethanol solution is in full contact with the container and the tool, and after cleaning, they should be thoroughly rinsed and dried to remove ethanol residues to avoid affecting subsequent sample processing.

[0041] In one embodiment of the present invention, the containers and tools need to be dried at 60°C to 80°C for 25min to 30min after cleaning to ensure that the containers and tools are dry and sterile.

[0042] In one embodiment of the present invention: in step 1 to step 6, the entire sample pretreatment process is carried out in an environment with a cleanliness level of 100, and the operator needs to wear disposable sterile protective clothing, gloves and masks that meet hygiene standards to ensure the sterility and safety of each operation. Example

[0043] Collect multiple coagulation samples, place them in specific sterile sample containers, perform visual inspection on the samples, remove samples with less than 2 ml of blood volume, coagulated samples (visible fibrin threads appear), and severely contaminated samples (visible foreign matter and color abnormalities appear), then randomly select 20 groups of samples that have undergone preliminary screening, transfer the samples after preliminary screening to a centrifuge device with temperature control function, then start the centrifuge device for centrifugal operation, centrifuge at 3000 rpm, centrifuge time is 15 min, and the centrifugation temperature is maintained at 5°C to separate the samples into a plasma layer and a blood cell layer, slowly draw out the separated plasma, transfer it to a new sterile container, add magnetic beads modified with ligands that specifically recognize impurities and interfering substances to the plasma, incubate at 25°C for 30 min, stir during incubation, and stir at a speed of 100 rpm. Adsorb the magnetic beads through an external magnetic field to remove impurities and interfering substances in the plasma, pass the plasma treated with magnetic beads through an ultrafiltration device, use an ultrafiltration membrane with a pore size of 0.22 μm, and perform centrifugation at a pressure of 0.1 MPa. Ultrafiltration was performed to remove macromolecules with a molecular weight greater than 100 kDa and small molecule interferences with a molecular weight less than 1 kDa in the plasma. The flux of the ultrafiltration membrane was monitored regularly during the ultrafiltration process. When the flux dropped to 50% of the initial flux, the ultrafiltration membrane needed to be replaced immediately. Subsequently, a 10% trichloroacetic acid solution was added to the ultrafiltered plasma. The solution was reacted for 20 minutes at a pH of 4.5 and a temperature of 40°C to precipitate trace impurities. The solution was placed in a centrifuge to remove the precipitate. The centrifuge speed was 3000 rpm and the solution was centrifuged for 10 minutes. After the chemical precipitation centrifugation reaction is completed, high performance liquid chromatography (HPLC) is used to detect the effect of precipitation removal to ensure that the plasma meets the quality requirements of the coagulation test after pretreatment. The concentration of the target substance in the treated plasma is determined by a spectrophotometer. According to the requirements of the coagulation test, when the concentration exceeds 5.0 mg / ml, it is diluted with 0.9% saline at a ratio of 1:1. When the concentration is lower than 0.5 mg / ml, it is concentrated by vacuum rotary evaporation to keep the plasma concentration between 0.5 mg / ml and 5.0mg / ml, the diluted and concentrated plasma needs to be mixed again, and the mixing method is vortex mixing for 3 minutes to ensure that the concentration gradient is broken and fully mixed. After each of the above operations is completed, the container and operating tools are cleaned with 75% ethanol solution to remove residual samples and processing reagents. The cleaning operation should follow strict operating procedures to ensure that the ethanol solution is in full contact with the container and tools, and thoroughly rinse and dry after cleaning to remove ethanol residues. The containers and tools must be dried at 60℃ for 30 minutes after cleaning to ensure that the containers and tools are dry and sterile. The above sample pretreatment process is carried out in an environment with a cleanliness level of 100. The operator must wear disposable sterile protective clothing, gloves and masks that meet hygiene standards, and then the treated samples are put into the coagulation tester for testing. . Example

[0044] Collect multiple coagulation samples, place them in specific sterile sample containers, perform visual inspection on the samples, remove samples with less than 2 ml of blood, coagulated samples (visible fibrin threads appear), and severely contaminated samples (visible foreign matter and color abnormalities appear), and then randomly select 20 groups of samples that have undergone preliminary screening. Transfer the samples after preliminary screening to a centrifuge with temperature control function, then start the centrifuge for centrifugation at 3000 rpm for 15 minutes. Maintain the centrifugation temperature at 15°C to separate the samples into a plasma layer and a blood cell layer, and slowly draw out the separated plasma. Transfer it to a new sterile container, add magnetic beads modified with ligands that specifically recognize impurities and interfering substances to the plasma, incubate it at 25°C for 30 minutes, stir it during the incubation process, and the stirring speed is 50 rpm. The magnetic beads are adsorbed by an external magnetic field to remove impurities and interfering substances in the plasma. The plasma treated with magnetic beads is passed through an ultrafiltration device, using an ultrafiltration membrane with a pore size of 0.22 μm and ultrafiltration at a pressure of 0.1 MPa to remove macromolecules with a molecular weight greater than 100 kDa and small molecule interfering substances with a molecular weight less than 1 kDa in the plasma. The flux of the ultrafiltration membrane is monitored regularly during the ultrafiltration process. When the flux drops to the initial When the flux reaches 50%, the ultrafiltration membrane needs to be replaced immediately, and then a 10% trichloroacetic acid solution is added to the ultrafiltered plasma. The reaction is carried out at a pH of 4.5 and a temperature of 45°C for 20 minutes to allow trace impurities to form a precipitate. The precipitate is removed in a centrifuge at a speed of 3000 rpm for 10 minutes. After the chemical precipitation centrifugation reaction is completed, the effect of precipitate removal is detected by high performance liquid chromatography (HPLC) to ensure that the plasma meets the quality requirements of the coagulation test after pretreatment. The concentration of the target substance in the treated plasma is determined by a spectrophotometer. According to the requirements of the coagulation test, each of the above operations is completed. After completion, the containers and operating tools are cleaned with 75% ethanol solution to remove residual samples and processing reagents. The cleaning operation should follow strict operating procedures to ensure that the ethanol solution is in full contact with the containers and tools, and they should be thoroughly rinsed and dried after cleaning to remove ethanol residues. The containers and tools must be dried at 60°C for 30 minutes after cleaning to ensure that the containers and tools are dry and sterile. The above sample pretreatment process is carried out in an environment with a cleanliness level of 100. Operators must wear disposable sterile protective clothing, gloves and masks that meet hygiene standards, and then the processed samples are placed in a coagulation tester for testing.

[0045] Control group,

[0046] Multiple coagulation samples were collected and placed in specific sterile sample containers. The samples were visually inspected and samples with less than 2 ml of blood, coagulated samples (visible fibrin threads appeared) and samples that were severely contaminated (visible foreign matter and color abnormalities appeared) were removed. Then, 20 groups of samples that had undergone preliminary screening were randomly selected and the processed samples were placed in a coagulation tester for testing.

[0047] Take 20 groups of samples from Example 1, Example 2 and the control group and two coagulation testers of the same specifications, stick labels of corresponding categories on the surfaces of 60 groups of sample containers for marking, and perform prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB) content and thrombin time (TT) tests on 20 samples in each of the three groups, and repeat the test three times for each sample on each coagulation tester, and record the test values.

[0048] According to the attached Figure 1 As shown, after the prothrombin time (PT), activated partial thromboplastin time (APTT) and thrombin time (TT) tests of the 20 groups of samples pretreated in Example 1, it was found that the pretreated samples had a shorter test time in the coagulation tester, a higher fibrinogen (FIB) content, and could more accurately evaluate the coagulation function. In addition, the difference in the average values ​​measured by two coagulation testers of the same specifications was less, making the test results more accurate. In summary, the 20 groups of samples pretreated in Example 1 can greatly improve the detection efficiency of the samples in the coagulation tester, greatly shorten the detection time, and can enhance the detection accuracy and reduce errors.

[0049] At the same time, the present application uses specific words to describe the embodiments of the present application. For example, "one embodiment", "an embodiment", and / or "some embodiments" refer to a certain feature, structure or characteristic related to at least one embodiment of the present application. Therefore, it should be emphasized and noted that "one embodiment" or "an embodiment" or "an alternative embodiment" mentioned twice or more in different positions in this specification does not necessarily refer to the same embodiment. In addition, certain features, structures or characteristics in one or more embodiments of the present application can be appropriately combined.

[0050] Some aspects of the present application may be performed entirely by hardware, entirely by software (including firmware, resident software, microcode, etc.), or by a combination of hardware and software. The above hardware or software may be referred to as "data blocks", "modules", "engines", "units", "components" or "systems". The processor may be one or more application specific integrated circuits (ASICs), digital signal processors (DSPs), digital signal processing devices (DAPDs), programmable logic devices (PLDs), field programmable gate arrays (FPGAs), processors, controllers, microcontrollers, microprocessors, or combinations thereof. In addition, various aspects of the present application may be represented as computer products located in one or more computer-readable media, which include computer-readable program codes. For example, computer-readable media may include, but are not limited to, magnetic storage devices (e.g., hard disks, floppy disks, tapes ...), optical disks (e.g., compact disks CDs, digital versatile disks DVDs ...), smart cards, and flash memory devices (e.g., cards, sticks, key drives ...).

[0051] A computer-readable medium may include a propagated data signal containing computer program code, such as in baseband or as part of a carrier wave. The propagated signal may have a variety of manifestations, including electromagnetic, optical, etc., or a suitable combination. A computer-readable medium may be any computer-readable medium other than a computer-readable storage medium, which may be connected to an instruction execution system, device or apparatus to communicate, propagate or transmit a program for use. The program code on the computer-readable medium may be propagated through any suitable medium, including radio, cable, fiber optic cable, radio frequency signal, or similar medium, or a combination of any of the above mediums.

[0052] Similarly, it should be noted that in order to simplify the description disclosed in this application, so as to help understand one or more embodiments of the invention, in the above description of the embodiments of the present application, sometimes multiple features are merged into one embodiment, drawings or descriptions thereof. However, this disclosure method does not mean that the features required by the object of the present application are more than the features mentioned in the claims. In fact, the features of the embodiment are less than all the features of the single embodiment disclosed above. In some embodiments, numbers describing the number of components and attributes are used. It should be understood that such numbers used in the description of the embodiments are modified by the modifiers "approximately", "approximately" or "substantially" in some examples. Unless otherwise specified, "approximately", "approximately" or "substantially" indicate that the number allows a variation of ±20%. Accordingly, in some embodiments, the numerical parameters used in the specification and claims are approximate values, which may change according to the characteristics required by individual embodiments. In some embodiments, the numerical parameters should take into account the specified significant digits and adopt the general method of retaining the digits. Although the numerical domains and parameters used to confirm the breadth of their range in some embodiments of the present application are approximate values, in specific embodiments, such numerical settings are as accurate as possible within the feasible range.

[0053] Although the present invention is disclosed as above in terms of preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art may make possible changes and modifications without departing from the spirit and scope of the present invention. Therefore, any modification, equivalent change and modification made to the above embodiments according to the technical essence of the present invention without departing from the content of the technical solution of the present invention shall fall within the scope of protection defined by the claims of the present invention.

Claims

1. A multi-sample processing method for a coagulation tester, characterized in that: The specific operation steps of the multi-sample processing method of the coagulation tester are as follows: Step 1: Collect multiple coagulation blood samples, place them in sterile sample containers, perform visual inspection on the samples, and remove samples with less than 2 ml of blood, coagulated samples, and severely contaminated samples; Step 2: Transfer the sample after preliminary screening to a centrifuge device with temperature control function, and then start the centrifuge device to perform centrifugal operation to separate the sample into a plasma layer and a blood cell layer; Step 3: Slowly draw out the separated plasma and transfer it to a new sterile container, add magnetic beads with surface modification that specifically recognize impurities and interfering substances to the plasma, and incubate at 23°C to 28°C for 30 to 35 minutes. Stir during the incubation process to adsorb the magnetic beads through an external magnetic field, thereby removing impurities and interfering substances in the plasma; Step 4: The plasma treated with magnetic beads is passed through an ultrafiltration device, using an ultrafiltration membrane with a pore size of 0.22 μm and ultrafiltration at a pressure of 0.1 MPa to remove macromolecules with a molecular weight greater than 100 kDa and small molecule interferences with a molecular weight less than 1 kDa in the plasma; Step 5: Add a 10% trichloroacetic acid solution to the ultrafiltered plasma, react for 18 to 22 minutes at a pH of 4.5 to 5.0 and a temperature of 37°C to 40°C to precipitate trace impurities, place in a centrifuge to remove the precipitate, and centrifuge at a speed of 3000 rpm to 3200 rpm for 8 to 12 minutes; Step 6: Use a spectrophotometer to measure the concentration of the target substance in the treated plasma. According to the requirements of the coagulation test, when the concentration exceeds 5.0 mg / ml, it is diluted with 0.9% saline at a ratio of 1:

1. When the concentration is lower than 0.5 mg / ml, it is concentrated by vacuum rotary evaporation to keep the plasma concentration between 0.5 mg / ml and 5.0 mg / ml. The treated sample is then placed in a coagulation tester for testing.

2. The multi-sample processing method of a coagulation tester according to claim 1, characterized in that: In the step 2, the centrifugal device used has a temperature control function, and the centrifugal treatment is performed at a speed of 3000 rpm to 3200 rpm, the centrifugal time is 13 min to 16 min, and the centrifugal temperature is maintained at 4°C to 6°C. This temperature can reduce the activity of enzymes in the blood and reduce the occurrence of biochemical reactions, thereby maintaining the original state of various components in the sample.

3. The multi-sample processing method of a coagulation tester according to claim 1, characterized in that: In the step three, the magnetic beads are incubated with slight stirring at a speed of 90 rpm to 110 rpm, which can promote uniform distribution of the magnetic beads in the plasma, avoid local aggregation of the magnetic beads, and increase the chance and efficiency of binding of the magnetic beads to impurities.

4. The multi-sample processing method of a coagulation tester according to claim 1, characterized in that: In the step 4, the flux of the ultrafiltration membrane is regularly monitored during the ultrafiltration process. When the flux drops to 50% of the initial flux, the ultrafiltration membrane needs to be replaced immediately.

5. The multi-sample processing method of a coagulation tester according to claim 1, characterized in that: In the step 5, after the chemical precipitation centrifugation reaction is completed, high performance liquid chromatography (HPLC) is used to detect the effect of precipitation removal to ensure that the plasma meets the quality requirements of the coagulation test after pretreatment.

6. The multi-sample processing method of a coagulation tester according to claim 1, characterized in that: In step six, the diluted and concentrated plasma needs to be mixed again by vortex mixing for 2 to 4 minutes to ensure that the concentration gradient is broken and the plasma is fully mixed.

7. The multi-sample processing method of a coagulation tester according to claim 1, characterized in that: After each step from step 1 to step 6 is completed, the container and operating tools are cleaned with 75% ethanol solution to remove residual samples and processing reagents. The cleaning operation should follow strict operating procedures to ensure that the ethanol solution is in full contact with the container and tools, and thoroughly rinse and dry after cleaning to remove ethanol residues and avoid affecting subsequent sample processing.

8. The multi-sample processing method of the coagulation tester according to claim 7, characterized in that: After cleaning, the containers and tools need to be dried at 60° C. to 80° C. for 25 min to 30 min to ensure that the containers and tools are dry and sterile.

9. The multi-sample processing method of a coagulation tester according to claim 1, characterized in that: In steps 1 to 6, the entire sample pretreatment process is carried out in an environment with a cleanliness level of 100, and operators are required to wear disposable sterile protective clothing, gloves and masks that meet hygiene standards to ensure the sterility and safety of each operation.

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