Blood coagulation system analyzer, blood coagulation system measuring device, blood coagulation system measuring system, blood coagulation system analyzing method, and blood coagulation system measuring method

Abstract

To provide a technique capable of accurately evaluating the blood coagulation ability due to a blood coagulation factor targeted by a medicine which is an object of the medicinal effect evaluation.SOLUTION: A blood coagulation system analysis device is given in which with respect to a blood sample to which an inhibitor inhibiting Xa factor and / or thrombin, and one or more reagents selected from X factor activator, prothrombin activator, and Xa factor are added, an analysis part is provided to analyze influence of the inhibitor to the blood coagulation system by using secular change data of electrical characteristic measured in the specific frequency or frequency band.SELECTED DRAWING: Figure 2

Description

[Technical field]

[0001] The present technology relates to a blood coagulation system analyzer, a blood coagulation system measuring device, a blood coagulation system measuring system, a blood coagulation system analyzing method, and a blood coagulation system measuring method. [Background technology]

[0002] In recent years, the use of direct oral anticoagulants (DOACs) has been increasing. Compared to warfarin, DOACs have a faster onset of action, less intracranial bleeding, and no dietary restrictions, so their use has been increasing in recent years as an alternative to warfarin.

[0003] On the other hand, the dosage must be reduced for elderly patients and those with renal impairment, and attention must be paid to the rise in blood DOACs concentration beyond the therapeutic range. In addition, when performing invasive procedures such as surgery, it is necessary to suspend the drug during planned surgery, but if complications such as intracranial bleeding or gastrointestinal bleeding or traumatic bleeding occur while the drug's effectiveness remains, the bleeding itself may become severe, making it difficult to stop bleeding even in emergency surgery. In such cases, it may become necessary to administer large amounts of fresh frozen plasma or administer off-label coagulation factor preparations (prothrombin complex preparations).

[0004] Although reversal drugs for DOACs have been developed in some countries, it is considered essential to evaluate blood coagulation ability when using reversal drugs to determine the indication for intervention and to evaluate the efficacy of DOACs. For example, the maximum blood concentration (Cmax) after one dose of the anti-factor X direct inhibitors edoxaban, apixaban, and rivaroxaban at the usual prescribed dose is 355ng / mL, 176ng / mL, and 348ng / mL, respectively. In addition, as a guideline for the period usually set for the drug holiday period, as proposed by Rosencher et al., if the time taken to eliminate the drug from the blood is twice the half-life, the cutoffs for avoiding the risk of bleeding are expected to be 89ng / mL, 44ng / mL, and 87ng / mL, which correspond to 25% of Cmax, respectively.

[0005] Common blood coagulation tests include the prothrombin time international normalized ratio (PT-INR) and activated partial thromboplastin time (APTT). These methods analyze the coagulation reactivity of proteins involved in the coagulation reaction, which are contained in the plasma obtained by centrifuging a blood sample.

[0006] These test methods are used to evaluate the function tests of extrinsic and intrinsic coagulation functions. In these tests, substances that induce extrinsic and intrinsic coagulation reactions are added in large excess so that test results can be obtained in a short time. These tests are performed using plasma obtained by centrifuging a blood sample, but cellular components such as platelets and red blood cells that play an important role in blood coagulation reactions in the body are removed by centrifugation, so there are many cases where the test results are inconsistent with the actual clinical pathology.

[0007] Other functional tests include thromboelastography and thromboelastometry, which are commercialized as TEG (registered trademark) and ROTEM (registered trademark), respectively. Thromboelastography is a method in which a blood sample is vibrated to measure the resonance frequency, and the increase in resonance frequency associated with an increase in viscoelasticity is measured. Thromboelastometry is a method in which a rotating pin is brought into contact with a blood sample filled in a cup, and the shear stress generated by the pin is evaluated as viscoelasticity. Therefore, it has been pointed out that there is a drawback in that measurements are performed while applying stress to immature blood clots in the process of clot formation, and no promise has been found for DOACs monitoring.

[0008] In addition, in recent years, a method of performing dielectric measurement during the blood coagulation process has been proposed as another method for easily and accurately evaluating blood coagulation measurement (e.g., Patent Documents 1 and 2). In this method, a blood sample is filled into a capacitor-like sample section consisting of a set of electrode pairs, and an AC electric field is applied to it to measure the change in complex dielectric constant associated with the coagulation process of the blood sample. Non-Patent Document 1 shows that the coagulation and fibrinolysis processes can be easily monitored by using this method. [Prior art documents] [Patent documents]

[0009] [Patent Document 1] JP 2010-181400 A [Patent Document 2] JP 2012-194087 A [Non-patent literature]

[0010] [Non-Patent Document 1] Y. Hayashi et al., Analytical Chemistry 87(19), 10072-10079 (2015) Summary of the Invention [Problem to be solved by the invention]

[0011] The blood coagulation reaction is a chain reaction called the blood coagulation cascade, as shown in Figure 1. Specifically, when the coagulation reaction starts, the inactive blood coagulation factors present in the blood are converted to active forms, which in turn activate the next inactive blood coagulation factor, thereby progressing the coagulation reaction.

[0012] As mentioned above, a method for easily and accurately evaluating blood coagulation measurement is to perform dielectric measurement during the blood coagulation process, which uses tissue factor (activation of the extrinsic coagulation process) or ellagic acid (activation of the intrinsic coagulation process) to initiate the coagulation reaction. In other words, this method is a method for measuring the degree of blood coagulation by activating factors upstream of the blood coagulation cascade to advance the blood coagulation reaction.

[0013] On the other hand, when evaluating blood coagulation ability for the purpose of evaluating the efficacy of anticoagulants such as DOACs, it is necessary to evaluate blood coagulation ability caused by factors such as Xa and thrombin, which are targets of anticoagulants such as DOACs. However, conventional methods activate factors upstream of the blood coagulation cascade, which results in a problem that downstream reactions of the blood coagulation cascade, such as Xa and thrombin, which are targets of anticoagulants such as DOACs, are highly dependent on the upstream coagulation activity.

[0014] Therefore, the main objective of this technology is to provide a technology capable of evaluating with high accuracy the blood coagulation function caused by the blood coagulation factor targeted by a drug that is the subject of efficacy evaluation. [Means for solving the problem]

[0015] The present inventors have conducted intensive research into technology capable of evaluating with high accuracy the blood coagulation ability caused by the blood coagulation factor targeted by a drug subject to efficacy evaluation. As a result, they have discovered that by using a reagent that activates the precursor of the coagulation factor on which the drug subject to efficacy evaluation acts, it is possible to eliminate the effects of coagulation activity upstream of the precursor, and have thus completed the present technology.

[0016] That is, the present technology comprises an inhibitor that inhibits factor Xa and / or thrombin, one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa; For the blood samples spiked with The present invention provides a blood coagulation system analyzer having an analysis unit that analyzes the effect of the inhibitor on the blood coagulation system by utilizing data on changes over time in electrical characteristics measured at a specific frequency or frequency band. As the factor X activator, a factor X activating enzyme derived from snake venom can be used. The blood coagulation system analyzer according to the present technology may include a reagent addition amount calculation unit that calculates the amount of reagent to be added. The reagent addition amount calculation unit can calculate the amount of the reagent to be added based on the time-course change data obtained from the blood sample of a healthy subject. The reagent addition amount calculation unit can calculate the amount of reagent to be added to be 0.5 to 2 when the amount of reagent to be added is 1 when the time-course data obtained from the blood sample of the healthy subject is in a predetermined range.

[0017] In the present technology, a first blood sample to which one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa have been added is subjected to a first time-dependent change data of an electrical property measured at a specific frequency or frequency band; A second time-dependent change data of the electrical property measured at a particular frequency or frequency band for a second blood sample to which the reagent has not been added; and The present invention provides a blood coagulation system analyzer having an analysis unit that utilizes the above-mentioned reagent to analyze the effects of coagulation factors upstream of the coagulation factor on which the reagent acts. The analysis section can analyze the influence of a coagulation factor upstream of the coagulation factor on which the reagent acts by correcting the second time-course data with the first time-course data.

[0018] In this technology, a blood sample to which an anticoagulant and a reagent that activates the precursor of a coagulation factor on which the anticoagulant acts have been added is subjected to the following steps: The present invention provides a blood coagulation system analyzer including an analysis unit that analyzes the effect of the anticoagulant on the blood coagulation system by utilizing data on changes over time in electrical characteristics measured at a specific frequency or frequency band.

[0019] The present technology comprises an inhibitor that inhibits factor Xa and / or thrombin, one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa; A measurement unit that measures electrical characteristics at a specific frequency or frequency band of a blood sample to which an analysis unit that analyzes the effect of the inhibitor on the blood coagulation system using data on the change over time of the measured electrical characteristics; The present invention provides a blood coagulation system measuring device comprising: The present technology also includes an inhibitor that inhibits factor Xa and / or thrombin; one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa; A measuring device for measuring electrical characteristics at a specific frequency or frequency band of a blood sample to which has been added; an analysis device that analyzes the effect of the inhibitor on the blood coagulation system using data on the change over time of the measured electrical characteristics; The present invention provides a blood coagulation system measuring system comprising:

[0020] In the present technology, a factor X activator, a prothrombin activator, and a factor Xa activator are used. a first blood sample having one or more reagents added thereto; A measurement unit that measures electrical characteristics at a specific frequency or frequency band for a second blood sample to which the reagent has not been added; A first time-dependent change data of an electrical property measured from the first blood sample; and second time-dependent change data of the electrical property measured from the second blood sample; and an analysis unit that uses the above-mentioned reagent to analyze the effect of a coagulation factor upstream of the coagulation factor on which the reagent acts; The present invention provides a blood coagulation system measuring device comprising: The present technology also includes a first blood sample to which one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa have been added; a measuring device for measuring an electrical characteristic at a specific frequency or frequency band of a second blood sample to which the reagent has not been added; A first time-dependent change data of an electrical property measured from the first blood sample; and second time-dependent change data of the electrical property measured from the second blood sample; and an analysis device for analyzing the influence of a coagulation factor upstream of the coagulation factor on which the reagent acts, using the The present invention provides a blood coagulation system measuring system comprising:

[0021] The present technology comprises an inhibitor that inhibits factor Xa and / or thrombin, one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa; For the blood samples spiked with The present invention provides a blood coagulation system analysis method that performs an analysis step of analyzing the effect of the inhibitor on the blood coagulation system by utilizing data on changes over time in electrical properties measured at a specific frequency or frequency band.

[0022] In the present technology, a first blood sample to which one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa have been added is subjected to a first time-dependent change data of an electrical property measured at a specific frequency or frequency band; A second time-dependent change data of the electrical property measured at a particular frequency or frequency band for a second blood sample to which the reagent has not been added; and The present invention provides a blood coagulation system analysis method that utilizes a reagent to perform an analysis step of analyzing the effects of coagulation factors upstream of the coagulation factor on which the reagent acts.

[0023] The present technology comprises an inhibitor that inhibits factor Xa and / or thrombin, one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa; A measuring step of measuring an electrical characteristic at a specific frequency or frequency band of the blood sample to which the an analysis step of analyzing the effect of the inhibitor on the blood coagulation system using data on the change over time of the measured electrical properties; The present invention provides a method for measuring the blood coagulation system.

[0024] The present technology includes a first blood sample to which one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa have been added; a measuring step of measuring electrical characteristics at a specific frequency or frequency band for a second blood sample to which the reagent has not been added; A first time-dependent change data of an electrical property measured from the first blood sample; and second time-dependent change data of the electrical property measured from the second blood sample; and and analyzing the effect of a coagulation factor upstream of the coagulation factor on which the reagent acts by utilizing the above-mentioned. and, The present invention provides a method for measuring the blood coagulation system.

[0025] In the present technology, the term "blood sample" refers to a sample containing red blood cells and liquid components such as plasma, and is not limited to blood itself. More specifically, examples of the blood sample include whole blood, plasma, or liquid samples containing blood components such as diluents and / or pharmaceutical additives. [Brief description of the drawings]

[0026] [Figure 1] FIG. 1 shows a schematic diagram of the blood coagulation cascade. [Diagram 2] FIG. 1 is a block diagram showing an example of an embodiment of a blood coagulation system analyzer 1 according to the present technology. [Diagram 3] FIG. 1 is a block diagram showing an example of an embodiment of a blood coagulation system analysis system 10 according to the present technology. [Figure 4] FIG. 1 is a block diagram showing an example of an embodiment of a blood coagulation system measuring device 2 according to the present technology. [Diagram 5] FIG. 1 is a block diagram showing an example of an embodiment of a blood coagulation system measuring system 20 according to the present technology. [Figure 6] FIG. 6 is a conceptual diagram showing an example different from FIG. 5 of the embodiment of the blood coagulation system measuring system 20 according to the present technology. [Figure 7] FIG. 7 is a block diagram showing an example of the embodiment of the blood coagulation system measuring system 20 according to the present technology, which is different from the examples shown in FIGS. 5 and 6. [Figure 8] 1 is a flowchart showing an example of an embodiment of a blood coagulation system analysis method according to the present technology. [Figure 9] 1 is a flowchart showing an example of an embodiment of a blood coagulation system measuring method according to the present technology. [Figure 10] FIG. 2 is a schematic cross-sectional view showing an example of an embodiment of a blood sample holder 211. [Figure 11] This is a typical graph showing the time course of data from a dielectric coagulometer measurement at 1 MHz. [Figure 12] 1 is a graph plotting the change in DBCM-CT versus DOACs (rivaroxaban or edoxaban) concentration when snake venom RVV-X was added in a volume of 10 μL per 180 μL of blood in Experimental Example 1. [Figure 13] 1 is a graph plotting the change in DBCM-CT versus DOACs (rivaroxaban or edoxaban) concentration when snake venom RVV-X was added in a volume of 20 μL per 180 μL of blood in Experimental Example 1. [Figure 14] 1 is a graph plotting the change in DBCM-CT versus DOACs (edoxaban) concentration when snake venom RVV-X was added in a volume of 40 μL per 180 μL of blood in Experimental Example 1. [Figure 15] 1 is a graph showing DBCM-CT values ​​versus blood concentration of rivaroxaban in Experimental Example 2. [Figure 16]This is a graph showing the results of measuring the clotting time at various concentrations of apixaban or rivaroxaban using TEG-6S (registered trademark) or ROTEM (registered trademark), which are blood coagulation test devices that measure blood viscoelasticity. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0027] Hereinafter, preferred embodiments of the present technology will be described with reference to the drawings. The embodiment described below is an example of a typical embodiment of the present technology, and is not intended to narrow the scope of the present technology. The description will be given in the following order. 1. Main issues and basic concept of this technology 2. Blood coagulation system analyzer 1, blood coagulation system analysis system 10 (1) Signal Acquisition Unit 11 (2) Analysis section 12 (3) Reagent addition amount calculation unit 13 (4) Output section 14 (5) Storage unit 15, storage device 105 (6) Display section 16, display device 106 (7) User Interface 17,107 3. Blood coagulation system measuring device 2, blood coagulation system measuring system 20 (1) Measuring unit 21, measuring device 201 (1-1) Blood sample holder 211 (1-2) A pair of electrodes 212a, 212b (1-3) Application unit 213 (1-4) Measuring mechanism (2) Drug Addition Section 22 (3) Control unit 23, control device 203 4. Blood coagulation system analysis method, blood coagulation system measurement method 5. Computer Programs

[0028] 1. Main issues and basic concept of this technology As mentioned above, when evaluating blood coagulation ability for the purpose of evaluating the efficacy of anticoagulants such as DOACs, it is necessary to evaluate blood coagulation ability caused by Xa factor, thrombin, etc., which are targets of anticoagulants such as DOACs. However, the conventional blood coagulation measurement method using tissue factor (activation of extrinsic coagulation process) or ellagic acid (activation of intrinsic coagulation process) to start the coagulation reaction is a method of measuring the degree of blood coagulation by progressing the blood coagulation reaction by activating factors upstream of the blood coagulation cascade, so that it is difficult to accurately evaluate blood coagulation ability caused by Xa factor, thrombin, etc., which are targets of anticoagulants such as DOACs, due to the large influence of the coagulation activity upstream of the blood coagulation cascade.

[0029] On the other hand, in this technology, by using a reagent that activates the precursor of the coagulation factor on which the drug targeted for efficacy evaluation acts, it is possible to eliminate the influence of coagulation activity upstream of the precursor.

[0030] Specifically, for example, when confirming the efficacy evaluation of anticoagulants such as DOACs, by adding a reagent that activates factor Xa, which is the target of anticoagulants such as DOACs, and factor X or prothrombin, which is a precursor of thrombin, to a blood sample, the influence of the coagulation activity upstream of factor X can be eliminated. As a result, the blood coagulation ability caused by factor Xa and thrombin, which are the targets of anticoagulants such as DOACs, can be evaluated with high accuracy. In this case, the same effect can be obtained by adding factor Xa itself, which is the target of anticoagulants such as DOACs, to a blood sample.

[0031] In addition, by changing the viewpoint of this technology, for example, by using the evaluation results of blood coagulation ability obtained from a blood sample to which a reagent that activates factor X or prothrombin, which is a precursor of factor Xa or thrombin, or factor Xa itself has been added, it is possible to correct the evaluation results of blood coagulation ability obtained from a blood sample to which these reagents have not been added (a blood sample using tissue factor (activating the extrinsic coagulation process) or ellagic acid (activating the intrinsic coagulation process))., it is possible to accurately evaluate blood coagulation ability caused by coagulation factors upstream of factor X.

[0032] 2. Blood coagulation system analyzer 1, blood coagulation system analysis system 10 2 is a block diagram showing an example of an embodiment of the blood coagulation system analyzer 1 according to the present technology. The blood coagulation system analyzer 1 according to the present technology includes a signal acquisition unit 11, an analysis unit 12, and an output unit 14. In addition, it may also include a reagent addition amount calculation unit 13, a storage unit 15, a display unit 16, a user interface 17, and the like, as necessary.

[0033] The signal acquiring unit 11, the analyzing unit 12, the output unit 14, the reagent addition amount calculation unit 13, the memory unit 15, the display unit 16, the user interface 17, etc. may be provided within the analyzing device 1, as in the blood coagulation system analyzing device 1 shown in FIG. 2, or a blood coagulation system analyzing system 10 may be provided as shown in FIG. 3, in which an analyzing device 101 equipped with the signal acquiring unit 11, the analyzing unit 12, the output unit 14, and, if necessary, the reagent addition amount calculation unit 13, and, if necessary, a memory device 105, a display device 106, a user interface 107, etc. are connected via a network.

[0034] It is also possible to provide the analysis unit 12 or analysis device 101, the storage unit 15 or storage device 105, and the display unit 16 or display device 106 in a cloud environment and connect them to the measurement unit 21 or measurement device 201 described below via a network. In this case, it is also possible to store the analysis results in the analysis unit 12 or analysis device 101 in the storage unit 15 or storage device 105 on the cloud, and share various pieces of information stored in the storage unit 15 or storage device 105 among multiple users.

[0035] Each section and device will be described in detail below.

[0036] (1) Signal Acquisition Unit 11 The blood coagulation system analyzer 1 and the analysis device 101 of the blood coagulation system analysis system 10 according to the present technology may include a signal acquisition unit 11. The signal acquisition unit 11 acquires data on changes over time in electrical properties of a blood sample measured at a specific frequency or frequency band. For example, the signal acquisition unit 11 acquires data on changes over time in electrical properties of the blood sample measured by a measurement unit 21 or a measurement device 201 described below.

[0037] (2) Analysis section 12 The blood coagulation system analyzer 1 and the analyzer 101 of the blood coagulation system analysis system 10 according to the present technology include an analysis unit 12. The analysis unit 12 analyzes the effect of an anticoagulant on the blood coagulation system and the effect of a specific coagulation factor in the blood coagulation cascade. Specific analysis methods will be described below for each embodiment.

[0038] First Embodiment The analytical method according to the first embodiment is a method for analyzing the effect of an anticoagulant on the blood coagulation system using a blood sample to which an anticoagulant and a reagent that activates the precursor of the coagulation factor on which the anticoagulant acts have been added.

[0039] As mentioned above, in conventional blood coagulation measurements, the blood coagulation reaction is initiated by activating factors upstream of the blood coagulation cascade, such as tissue factor (activating the extrinsic coagulation process) or ellagic acid (activating the intrinsic coagulation process), which means that the measurement is highly dependent on upstream coagulation activity rather than on the blood coagulation factor targeted by the anticoagulant, making it difficult to evaluate blood coagulation ability resulting only from the blood coagulation factor targeted by the anticoagulant.

[0040] On the other hand, in the present technology, since the measurement is performed in a state where an anticoagulant and a reagent that activates the precursor of the coagulation factor on which the anticoagulant acts are added to the blood sample, the influence of the coagulation activity upstream of the precursor can be eliminated. As a result, the blood coagulation ability caused by the blood coagulation factor targeted by the drug to be evaluated for efficacy can be evaluated with high accuracy.

[0041] In this technique, it is possible to add tissue factor, an extrinsic coagulation activator, at a low concentration (for example, a final concentration of 1 pM or less, etc.) as in conventional blood coagulation measurements, but this is not essential when using a factor X activator or the like as a reagent, as described below, because the coagulation reaction is initiated more strongly by activation of factor X than by factor VII, which is related to tissue factor. On the other hand, adding tissue factor, an extrinsic coagulation activator, at a high concentration is not preferred because it may exhibit a non-negligible coagulation activity relative to the coagulation activation ability downstream of the blood coagulation cascade.

[0042] Examples of combinations of an anticoagulant and a reagent that activates a precursor of a coagulation factor on which the anticoagulant acts include a factor Xa inhibitor and a factor X activator, a thrombin inhibitor and a prothrombin activator, etc. When analyzing the effect of a factor Xa inhibitor on the blood coagulation system, the same effect can be obtained by using factor Xa itself.

[0043] Examples of inhibitors that inhibit factor Xa and / or thrombin include edoxaban, apixaban, rivaroxaban, dabigatran, and the like.

[0044] An example of an X factor activator is snake venom (Russell's viper venom) RVV-X.

[0045] Examples of prothrombin activators include factor Xa, thrombin-like enzymes derived from snake venom, and the like.

[0046] As will be shown in the examples described later, data on the change over time in electrical properties obtained from a blood sample to which an anticoagulant and a reagent that activates the precursor of the coagulation factor on which the anticoagulant acts have been added shows a correlation with the efficacy of the anticoagulant, and therefore, by utilizing the data on the change over time, the analysis unit 12 can analyze with high accuracy the effect of an inhibitor on the blood coagulation system.

[0047] <Second embodiment> The analytical method according to the second embodiment is a method for analyzing the effects of coagulation factors upstream of the coagulation factor acted upon by a first blood sample to which one or more reagents selected from a factor X activator, a prothrombin activator, and factor Xa have been added, and a second blood sample to which the reagents have not been added.

[0048] The data on the change over time of the electrical properties obtained from a first blood sample to which one or more reagents selected from a factor X activator, a prothrombin activator, and factor Xa have been added correlates with the blood coagulation ability caused by the coagulation factors downstream of the coagulation factors acting on by the reagents. Therefore, by correcting the data on the change over time of the electrical properties obtained from a second blood sample to which the reagents have not been added with the data on the change over time of the electrical properties obtained from the first blood sample, it is possible to analyze the influence of the coagulation factors upstream of the coagulation factors acting on by the reagents.

[0049] (3) Reagent addition amount calculation unit 13 The blood coagulation system analyzer 1 and the analysis device 101 of the blood coagulation system analysis system 10 according to the present technology may be provided with a reagent addition amount calculation unit 13 that calculates the amount of reagent to be added. In the present technology, the reagent addition amount calculation unit 13 is not essential, and for example, as described above, when the analysis unit 12 or the analysis device 101 is provided on the cloud, it is also possible to add reagent using the reagent addition amount calculated by another user.

[0050] The reagent addition amount calculation unit 13 can calculate the amount of the reagent to be added based on the time-course change data obtained from the blood sample of the healthy subject. More specifically, when the amount of the reagent to be added that brings the time-course change data obtained from the blood sample of the healthy subject into a predetermined range is set to 1, the amount of the reagent to be added can be calculated to be 0.5 to 2.

[0051] (4) Output section 14 The blood coagulation system analyzer 1 and the analysis device 101 of the blood coagulation system analysis system 10 according to the present technology can be provided with an output unit 14. The output unit 14 can output the analysis data analyzed in the analysis unit 12, the amount of reagent added calculated in the reagent addition amount calculation unit 13, and the like to a memory unit 15, a storage device 105, a display unit 16, a display device 106, a measurement unit 21, and a measurement device 201, which will be described later.

[0052] Furthermore, the output unit 14 can be configured to generate a notification signal at a specific time point and notify the user of the result in real time only when an abnormal analysis result is obtained during measurement by, for example, the measurement unit 21 or the measurement device 201 described below. This improves usability because the user is notified of the analysis result only at the specific time point when an abnormal analysis result is confirmed.

[0053] The method of notifying the user is not particularly limited, and can be, for example, via the display unit 16 described below, a display, a printer, a speaker, lighting, etc. Also, for example, the output unit 14 can be used in combination with a device having a communication function for sending an e-mail or the like to a mobile device such as a mobile phone or smartphone to notify that a notification signal has been generated.

[0054] (5) Storage unit 15, storage device 105 The blood coagulation system analyzer 1 and the blood coagulation system analysis system 10 according to the present technology may include a memory unit 15 and a storage device 105 for storing various data. The memory unit 15 and the storage device 105 can accumulate and store all kinds of data, such as various data acquired by the signal acquisition unit 11, analysis data analyzed by the analysis unit 12, the amount of reagent added calculated by the reagent addition amount calculation unit 13, various data related to blood coagulation system analysis, and various data related to blood coagulation system measurement, such as measurement data measured by the measurement unit 21 and the measurement device 201 described below.

[0055] The memory unit 15 and the storage device 105 are not essential for the blood coagulation system analyzer 1 and the blood coagulation system analysis system 10 according to the present technology, and each piece of data can be output from the output unit 14 to the outside of the device or system and stored in an external storage device. The storage device 105 can be provided in a cloud environment and can be connected to the blood coagulation system analyzer 1 and the blood coagulation system analysis system 10 according to the present technology via a network. In this case, various pieces of data stored in the storage device 105 on the cloud can be shared by multiple users.

[0056] A database can be constructed based on various data related to blood coagulation system analysis and various data related to blood coagulation system measurement described later in the memory unit 15 and the storage device 105. In this case, the analysis unit 12 and the reagent addition amount calculation unit 13 can also perform various analyses and calculations by referring to the database.

[0057] For example, by referring to analysis data and calculation data obtained in the past, measurement data obtained using external measuring devices 201A-201D as shown in FIG. 7 described later, and measurement data collected on other samples, it is possible to perform more accurate analysis and calculation.

[0058] The configuration of the storage unit 15 and the storage device 105 is not particularly limited, and for example, a hard disk drive, a flash memory, an SSD (Solid State Drive), or the like can be adopted.

[0059] In addition, in the present technology, the operating programs of the blood coagulation system analysis device 1 and the blood coagulation system analysis system 1 according to the present technology may be stored in the storage unit 15 or the storage device 105.

[0060] (6) Display section 16, display device 106 The blood coagulation system analyzer 1 and the blood coagulation system analysis system 10 according to the present technology can be provided with a display unit 16 and a display device 106 for displaying various data. The display unit 16 and the display device 106 can display all kinds of data, such as various data acquired by the signal acquisition unit 11, analysis data analyzed by the analysis unit 12, the amount of reagent added calculated by the reagent addition amount calculation unit 13, various data related to blood coagulation system analysis, and various data related to blood coagulation system measurement, such as measurement data measured by the measurement unit 21 and the measurement device 201 described below.

[0061] The configuration of the display unit 16 and the display device 106 is not particularly limited, and for example, a general display device such as a display or a printer can be used. Note that the display unit 16 and the display device 106 are not essential to the blood coagulation system analyzer 1 and the blood coagulation system analysis system 10 according to the present technology, and each data can be output from the output unit 14 to the outside of the device or system and displayed on an external display device.

[0062] (7) User Interface 17,107 The blood coagulation system analyzer 1 and the blood coagulation system analyzing system 10 according to the present technology may further include a user interface 17, 107 for user operation. The user can access each part or device through the user interface 17, 107 and operate each part or device.

[0063] In the present technology, the user interface 17, 107 is not essential, and an external operation device may be connected. As the user interface 17, 107, for example, a mouse, a keyboard, etc. may be used.

[0064] 3. Blood coagulation system measuring device 2, blood coagulation system measuring system 20 4 is a block diagram showing an example of an embodiment of the blood coagulation system measuring device 2 according to the present technology. The blood coagulation system measuring device 2 according to the present technology includes a measuring unit 21, a signal acquiring unit 11, an analyzing unit 12, and an output unit 14. In addition, as necessary, it may also include a reagent addition amount calculating unit 13, a memory unit 15, a display unit 16, a user interface 17, a drug adding unit 22, a control unit 23, and the like.

[0065] The measurement unit 21, analysis unit 12, reagent addition amount calculation unit 13, memory unit 15, display unit 16, user interface 17, etc. may be provided within the analysis device 1, as in the blood coagulation system measurement device 2 shown in Figure 4, or a blood coagulation system measurement system 20 may be provided in which an analysis device 101 equipped with a signal acquisition unit 11, analysis unit 12, output unit 14, and, if necessary, a reagent addition amount calculation unit 13, a measurement device 201, and, if necessary, a control device 203, a memory device 105, a display device 106, a user interface 107, etc. are connected via a network, as shown in Figure 5.

[0066] It is also possible to provide the analysis unit 12 or analysis device 101, the storage unit 15 or storage device 105, and the display unit 16 or display device 106 in a cloud environment and connect them to the measurement unit 21 or measurement device 201 via a network. In this case, the analysis results in the analysis unit 12 or analysis device 101 can be stored in the storage unit 15 or storage device 105 on the cloud, and various pieces of information stored in the storage unit 15 or storage device 105 can be shared by multiple users.

[0067] Fig. 6 is a conceptual diagram showing an example of the embodiment of the blood coagulation system measurement system 20 according to the present technology, which is different from Fig. 5. The blood coagulation system measurement system 20 according to the embodiment of Fig. 6 includes a measurement device 201 and a program described later for causing a computer to realize the same analysis as that performed by the analysis device 101 according to the present technology.

[0068] Fig. 7 is a block diagram showing an example of the embodiment of the blood coagulation system measuring system 20 according to the present technology, which is different from Fig. 5 and Fig. 6. The blood coagulation system measuring system 20 according to the embodiment of Fig. 7 is an example in which an analysis device 101 according to the present technology, a plurality of measuring devices 201A-D, and, if necessary, a storage device 105 are connected via a network.

[0069] 7, data on changes over time in electrical properties measured from blood samples by a plurality of measuring devices 201A-D can be analyzed by a single analyzing device 101. This allows the measuring data from the different measuring devices 201A-D to be referred to by the analyzing device 101, making it possible to perform more accurate analysis and calculations.

[0070] It is also possible to clearly divide the roles of each measuring device 201A, for example, to perform a measurement using a blood sample from a healthy subject in one measuring device 201A, and based on the results, to calculate the amount of reagent to be added in the reagent addition amount calculation unit 13 of the analysis section 101, and then to perform a measurement in another measuring device 201B using a blood sample from the patient to which the calculated amount of reagent has been added.

[0071] Each part and device will be described in detail below. Note that the signal acquisition unit 11, analysis unit 12, output unit 14, reagent addition amount calculation unit 13, memory unit 15, display unit 16, user interface 17, analysis device 101, storage device 105, display device 106, and user interface 107 are the same as those of the blood coagulation system analysis device 1 and blood coagulation system analysis system 10 described above, and therefore descriptions thereof will be omitted here.

[0072] (1) Measuring unit 21, measuring device 201 The measurement section 21 and the measurement device 201 at least include a blood sample holder 211 and a measurement mechanism. In addition, a pair of electrodes 212a, 212b and an application section 213 may be provided as necessary.

[0073] (1-1) Blood sample holder 211 The blood sample holding section 211 is a section that holds the blood sample B. In the blood coagulation system measuring device 2 and the blood coagulation system measuring system 20 according to the present technology, the form of the blood sample holding section 211 is not particularly limited, and the form is not specified as long as it is a form that allows the blood sample B to be held for a certain period of time when the electrical characteristics of the blood sample B are measured.

[0074] Fig. 10 is a schematic cross-sectional view showing an example of an embodiment of the blood sample holding section 211. The blood sample holding section 211 according to the embodiment of Fig. 10 comprises a blood sample holding container 211a and a container holder 211b that holds the blood sample holding container 211a.

[0075] The blood sample holding container 211a holds a blood sample B to be analyzed. In the blood coagulation system measuring device 2 and blood coagulation system measuring system 20 according to the present technology, electrical properties of the blood sample B are measured while the blood sample B is held in the blood sample holding container 211a. For this reason, it is preferable that the blood sample holding container 211a is configured to be able to be sealed while holding the blood sample B. However, it does not have to be configured to be airtight as long as it can be held for the time required to measure the electrical properties and does not affect the measurement.

[0076] A specific method for introducing the blood sample B into the blood sample holding container 211a and sealing the container is not particularly limited, and the blood sample B can be introduced by any suitable and free method depending on the form of the blood sample holding container 211a, etc. For example, a method in which a lid is provided on the blood sample holding container 211a, the blood sample B is introduced using a pipette or the like, and then the lid is closed to seal the container may be mentioned.

[0077] The form of the blood sample holding container 211a is not particularly limited as long as it can hold the blood sample B to be analyzed within the device, and can be freely designed as appropriate. The blood sample holding container 211a can be made up of one or more containers.

[0078] The specific shape of the blood sample holding container 211a is not particularly limited, and as long as it is capable of holding the blood sample B to be analyzed, it can be freely designed as appropriate depending on the state of the blood sample B, etc., such as a cylinder, a polygonal tube with a polygonal cross section (triangle, square or more), a cone, a polygonal pyramid with a polygonal cross section (triangle, square or more), or a combination of one or more of these.

[0079] Furthermore, the material constituting the blood sample holding container 211a is not particularly limited, and can be freely selected as appropriate within a range that does not affect the state of the blood sample B to be analyzed. In the present technology, it is particularly preferable that the blood sample holding container 211a is made of resin, from the viewpoint of ease of processing and molding. In the present technology, the type of resin that can be used is not particularly limited, and one or more types of resin that can be applied to holding the blood sample B can be freely selected as appropriate and used. For example, hydrophobic and insulating polymers, copolymers, blend polymers, etc., such as polypropylene, polymethyl methacrylate, polystyrene, acrylic, polysulfone, and polytetrafluoroethylene can be mentioned.

[0080] In the present technology, it is particularly preferable to form the blood sample holding container 211a from one or more resins selected from polypropylene, polystyrene, acrylic, and polysulfone. These resins have the property of having low coagulation activity for blood samples, and are therefore suitable for measuring blood samples.

[0081] In addition, in the present technology, a known disposable cartridge type may also be used as the blood sample holding container 211a.

[0082] In this technology, when various drugs and reagents are used, it is possible to previously store the predetermined drugs and reagents in the blood sample holding container 211a in a solidified or liquid form. For example, an anticoagulant such as an inhibitor that inhibits factor Xa and / or thrombin, a factor X activator, a prothrombin activator, a reagent such as factor Xa, etc., can be placed in the blood sample holding container 211a in advance. In this way, by previously storing drugs and reagents in the blood sample holding container 211a, the drug addition section 22 described below and a section for holding the drug and reagent are not required, and the device can be made smaller and the cost can be reduced. In addition, the user does not need to take the trouble of replacing drugs, and device maintenance of the drug addition section 22 and the section for holding the drug and reagent is not required, so that usability can be improved.

[0083] In the blood coagulation system measuring device 2 and blood coagulation system measuring system 20 according to the present technology, the number of blood sample holding containers 211a is not particularly limited, and one or more blood sample holding containers 211a can be freely arranged as appropriate depending on the amount, type, etc. of the blood sample B to be analyzed.

[0084] The container holder 211b holds the blood sample holding container 211a. The specific form of the container holder 211b is not particularly limited, and can be freely designed as appropriate as long as it can hold the blood sample holding container 211a containing the blood sample B to be analyzed.

[0085] The material constituting the container holding portion 211b is not particularly limited either, and can be freely selected as appropriate depending on the shape of the blood sample holding container 211a, etc.

[0086] In addition, in the present technology, the container holder 211b may have a function (such as a barcode reader) of automatically reading information about the blood sample holding container 211a from an information recording medium provided in the blood sample holding container 211a. Examples of the information recording medium include an IC card, an IC tag, a card provided with a barcode or a matrix type two-dimensional code, and a paper or a sticker on which a barcode or a matrix type two-dimensional code is printed.

[0087] (1-2) A pair of electrodes 212a, 212b The pair of electrodes 212a, 212b come into contact with the blood sample B during measurement and apply a required voltage to the blood sample B.

[0088] The arrangement and shape of the pair of electrodes 212a, 212b are not particularly limited, and may be freely designed as appropriate as long as it is possible to apply a voltage required for the blood sample B. In this embodiment, the pair of electrodes 212a, 212b are configured to be integrally molded with the blood sample holding container 211a, but are not limited to this, and may be configured such that the electrodes are inserted from the outside.

[0089] The material constituting the electrodes 212a, 212b is not particularly limited, and one or more known electrically conductive materials may be freely selected and used as appropriate within the scope of not affecting the state of the blood sample B to be analyzed. Specific examples include titanium, aluminum, stainless steel, platinum, gold, copper, graphite, etc.

[0090] In the present technology, it is particularly preferable to form the electrodes 212a, 212b from an electrically conductive material containing titanium among these. Titanium has the property of having low coagulation activity in blood samples, and is therefore suitable for measuring the blood sample B.

[0091] (1-3) Application unit 213 The application unit 213 applies an alternating voltage to the pair of electrodes 212a, 212b at a predetermined time interval. More specifically, for example, the application unit 213 applies an alternating voltage to the pair of electrodes 212a, 212b starting from a time point when the application unit 213 receives a command to start measurement or a time point when the power of the device 100 is turned on. More specifically, the application unit 213 applies an alternating voltage of a set frequency or a frequency controlled by the control unit 23 described later to the pair of electrodes 212a, 212b at each set measurement interval or each measurement interval controlled by the control unit 23 or the control device 203 described later.

[0092] (1-4) Measuring mechanism The measurement mechanism in the measurement unit 21 and the measurement device 201 measures electrical characteristics of a blood sample placed between a pair of electrodes 212a, 212b at a specific frequency or frequency band. Examples of electrical characteristics measured by the measurement unit 21 and the measurement device 201 include impedance, conductance, admittance, capacitance, dielectric constant, conductivity, phase angle, and quantities obtained by converting these into electrical quantities. Note that the blood coagulation system measurement device 2 and the blood coagulation system measurement system 20 according to the present technology can evaluate one of these electrical characteristics, but two or more electrical characteristics can also be used.

[0093] The configuration of the measurement mechanism in the measurement unit 21 and the measurement device 201 is not particularly limited, and can be appropriately set according to the electrical characteristics to be measured. For example, when applying an AC voltage between an electrode pair to measure the impedance or complex dielectric constant of blood, an impedance analyzer or a network analyzer can be used. Note that, in the present technology, measurements may be performed only for the frequency or frequency band used in the analysis unit 12 or the analysis device 101, but it is also possible to change the frequency to measure the electrical characteristics in a wide band and extract the frequency or frequency band to be used for evaluation from the obtained spectrum.

[0094] (2) Drug Addition Section 22 The blood coagulation system measuring device 2 and blood coagulation system measuring system 20 according to the present technology may be provided with a drug addition section 22. The drug addition section 22 automatically supplies one or more types of drugs (including reagents, the same applies below) to the measuring section 21 or the blood sample holding section 211 of the measuring device 201. In the blood coagulation system measuring device 2 and blood coagulation system measuring system 20 according to the present technology, this drug addition section 22 is not essential, but by providing the drug addition section 22, each step of blood coagulation system measurement can be performed automatically.

[0095] The drug addition unit 22 can supply a drug of a predetermined type and amount to the blood sample holding unit 211, but can also supply a drug of an amount calculated in the reagent addition amount calculation unit 13 to the blood sample holding unit 211.

[0096] The specific method of supplying the drug is not particularly limited, and for example, the drug can be automatically supplied to the blood sample holding container 211a of the blood sample holding unit 211 using a pipetter and a tip attached to the tip of the pipetter. In this case, it is preferable that the tip is disposable in order to prevent measurement errors and the like. The drug can also be automatically supplied to the blood sample holding container 211a from a drug storage tank using a pump or the like. Furthermore, it is also possible to automatically supply the drug to the blood sample holding container 211a using a permanently installed nozzle or the like. In this case, it is preferable that the nozzle is provided with a cleaning function in order to prevent measurement errors and the like.

[0097] In particular, the drug is preferably supplied in a manner that allows a constant amount of drug to be supplied without contacting the blood sample holding container 211a. For example, if the drug is in liquid form, it can be supplied by ejection. More specifically, for example, the drug liquid is introduced in advance into a discharge pipe, and the drug liquid can be ejected and supplied to the blood sample holding container 211a through a pipe connected to the discharge pipe by blowing pressurized air, which is connected separately, into the pipe for a short period of time. In this case, the amount of drug liquid ejected can be adjusted by adjusting the air pressure and the valve opening and closing time.

[0098] In addition to blowing in air, the liquid medicine can be discharged and supplied to the blood sample holding container 211a by utilizing the vaporization of the liquid medicine itself or the air dissolved therein by heating. In this case, the volume of bubbles generated can be adjusted by adjusting the time and voltage applied to a vaporization chamber in which a heating element or the like is installed, and the discharge amount of the liquid medicine can be adjusted.

[0099] Furthermore, it is also possible to supply the liquid medicine to the blood sample holding container 211a by driving a movable part provided in the pipeline using a piezoelectric element (piezo element) or the like without using air, and sending out an amount of the liquid medicine determined by the volume of the movable part. It is also possible to supply the medicine by using a so-called inkjet method, in which the liquid medicine is made into fine droplets and sprayed directly into the desired blood sample holding container 211a.

[0100] In addition, in the present technology, the drug adding section 22 can also be provided with a stirring function, a temperature control function, a function for identifying the type of drug and the like and automatically reading it (such as a barcode reader), and the like.

[0101] (3) Control unit 23, control device 203 The blood coagulation system measuring device 2 and blood coagulation system measuring system 20 according to the present technology may be provided with a control unit 23 and a control device 203 that control measurement conditions such as the measurement frequency, the temperature of the blood sample and the drug, the measurement time, the measurement interval, etc. In the present technology, the control unit 23 and the control device 203 are not essential, and it is also possible to control the measurement conditions using an external control device.

[0102] As an example of control of measurement conditions, for example, a specific method of measurement time control may involve controlling the measurement interval according to the amount of data required for the desired analysis, or controlling the timing of the end of measurement when the measurement value becomes almost flat.

[0103] Also, for example, specific methods for controlling the measurement frequency include changing the frequency of the AC voltage applied between the electrodes 212a and 212b, superimposing multiple frequencies to measure impedance at multiple frequencies, etc. Specific methods include arranging multiple single frequency analyzers in parallel, sweeping the frequency, superimposing the frequencies and extracting information on each frequency with a filter, measuring the response to an impulse, etc.

[0104] Furthermore, for example, a specific method of temperature control is to provide a temperature adjustment function to the area where the blood sample B is waiting, the area where the drug is waiting such as the drug addition section 22, or the blood sample holding section 211, thereby enabling temperature control of the blood sample B and the drug during measurement.

[0105] 4. Blood coagulation system analysis method, blood coagulation system measurement method 8 is a flow chart showing an example of an embodiment of the blood coagulation system analysis method according to the present technology. The blood coagulation system analysis method according to the present technology includes a signal acquisition step S11, an analysis step S12, and an output step S14. In addition, it is also possible to perform a reagent addition amount calculation step S13, a storage step S15, a display step S16, etc., as necessary.

[0106] 9 is a flowchart showing an example of an embodiment of the blood coagulation system measuring method according to the present technology. The blood coagulation system measuring method according to the present technology includes a measuring step S21, a signal acquiring step S11, an analyzing step S12, and an outputting step S14. In addition, it is also possible to perform a reagent addition amount calculating step S13, a storing step S15, a displaying step S16, a drug adding step S22, a control step S23, and the like, as necessary.

[0107] Since each step is the same as the step performed by each part of the blood coagulation system analyzer 1 and blood coagulation system measuring device 2 according to the present technology described above, the explanation will be omitted here.

[0108] 5. Computer Programs First Embodiment The computer program according to the first embodiment includes an inhibitor that inhibits factor Xa and / or thrombin; one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa; For the blood samples spiked with This is a program for enabling a computer to realize an analytical function for analyzing the effect of the inhibitor on the blood coagulation system by utilizing data on the change over time of electrical properties measured at a specific frequency or frequency band.

[0109] <Second embodiment> The computer program according to the second embodiment includes a first blood sample to which one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa have been added, the first blood sample being measured at a specific frequency or frequency band, and A second time-dependent change data of the electrical property measured at a particular frequency or frequency band for a second blood sample to which the reagent has not been added; and This is a program for enabling a computer to realize an analytical function for analyzing the effects of coagulation factors upstream of the factor on which the reagent acts, utilizing the above-mentioned.

[0110] <Third embodiment> The computer program according to the third embodiment executes the following for a blood sample to which an anticoagulant and a reagent that activates a precursor of a coagulation factor on which the anticoagulant acts have been added: This is a program for enabling a computer to realize an analytical function for analyzing the effect of the anticoagulant on the blood coagulation system by utilizing data on changes over time in electrical properties measured at a specific frequency or frequency band.

[0111] The computer program according to the present technology is recorded on an appropriate recording medium. The computer program according to the present technology can also be stored in a cloud environment or the like, and a user can download it to a personal computer or the like via a network for use. Note that the analysis function in the computer program according to the present technology is the same as the function performed by the analysis unit 12 of the blood coagulation system analyzer 1 according to the present technology described above, and therefore a description thereof will be omitted here. EXAMPLES

[0112] The present technology will be described in further detail below based on examples. It should be noted that the embodiment described below is merely an example of a typical embodiment of the present invention, and the scope of the present technology is not to be construed as being narrow.

[0113] <Experimental Example 1> [Specimen] Venous blood from 10 healthy volunteers was collected in a standard evacuated blood collection tube (containing 1 volume of 3.2% citric acid for every 9 volumes of blood) using citrate as an anticoagulant. Rivaroxaban or edoxaban, which are Xa inhibitors widely used as DOACs, were added at clinically important concentrations of 0 to 400 ng / mL.

[0114] [reagent] To overcome the anticoagulant effect of citrate, calcium chloride was used in the amount (2.4 μmol) normally used in dielectric coagulometry.

[0115] As an example of an X factor activator, snake venom (Russell's viper toxin) RVV-X was used. RVV-X was purchased from BioMedica Diagnostics, Inc. as a Lupus Anticoagulants test reagent, and the solution was dissolved according to the manufacturer's instructions and used at volumes of 10 μL, 20 μL, or 40 μL per 180 μL of blood.

[0116] These reagents were placed in advance in a dielectric coagulometer measurement cartridge and heated to 37° C. before measurement in the dielectric coagulometer device.

[0117] [measurement] The measurements were performed using a dielectric blood coagulometer (DBCM). When the blood collection tube was set in the device, it was heated to 37°C, and just before the start of the measurement, the blood was stirred by automatic pipetting and dispensed into the measurement cartridge. The dielectric spectrum data was continuously measured and recorded until the measurement was completed.

[0118] [analysis] The parameter extraction method used for the analysis is not particularly limited, but in this experimental example, we focused on data at 1 MHz. At frequencies around 1 MHz, as shown in Figure 11, bimodal characteristics are often observed. In this experimental example, the time at which the second peak is given is defined as DBCM-CT, but more generally, a characteristic time that correlates with the time at which the second peak is given can be used as the coagulation time measured by a blood coagulation system analyzer using dielectric constant changes. The graph in Figure 11 is a typical graph showing the time-series change in 1 MHz data in dielectric coagulometer measurement. The change is shown by dividing by the dielectric constant immediately after the start of measurement.

[0119] [result] Figures 12 to 14 are graphs plotting the change in DBCM-CT versus DOACs concentration when snake venom RVV-X was added in volumes of 10 μL, 20 μL, or 40 μL per 180 μL of blood.

[0120] As shown in Figure 12, at low concentrations of RVV-X (10 μL added per 180 μL of blood), DBCM-CT was generally extended with increasing DOACs concentration, but we found a problem in that the rise in DOACs concentration in the low DOACs concentration range of 0 to 50 ng / mL was slow, resulting in low sensitivity in the low DOACs concentration range.

[0121] Furthermore, as shown in Figure 14, it was found that at high concentrations of RVV-X (40 μL added per 180 μL of blood), the change in DBCM-CT plateaued at high DOACs concentrations, revealing the problem of low sensitivity at high DOACs concentrations.

[0122] On the other hand, Figure 13 shows the case of a medium concentration of RVV-X (20 μL added per 180 μL of blood), in which good DBCM-CT changes were obtained over the entire range of 0 to 400 ng / mL, which is the clinically important DOACs blood concentration.

[0123] From the above results, it was found that the amount of RVV-X reagent corresponding to Figure 13 or a concentration close to that is optimal for monitoring DOACs.

[0124] [Consideration] When the activity of a reagent such as a factor X activator changes depending on the source of the reagent or the difference between lots, there is a problem of how to practically determine the amount of reagent. To address this issue, when an anticoagulant such as DOACs is added to healthy blood, for example, for blood with an edoxaban concentration of 200 ng / mL, it is sufficient to determine the amount of reagent (or the reagent concentration, the reagent activity) that makes the DBCM-CT 400 to 500 seconds. When the amount of reagent (or the reagent concentration, the reagent activity) at this time is x, the amount of reagent (or the reagent concentration, the reagent activity) that deviates from the range of 0.5x to 2.0x is not suitable because the sensitivity decreases in the low concentration range or the high concentration range as shown in Figures 12 and 14. The optimal amount of reagent (or the reagent concentration, the reagent activity) is x, but it is presumed that it is also practical to use it in the range of about 0.7x to 1.7x around that.

[0125] <Experimental Example 2> [Specimen] Venous blood from 10 healthy volunteers was collected in the same vacuum blood collection tubes (blood collection tubes containing 1 volume of 3.2% citric acid per 9 volumes of blood) as in Experimental Example 1. Rivaroxaban, an Xa inhibitor widely used as a DOAC, was added at a clinically important concentration range of 0 to 400 ng / mL.

[0126] [reagent] To overcome the anticoagulant effect of citrate, calcium chloride was used in the amount (2.4 μmol) normally used in dielectric coagulometry.

[0127] Snake venom (Russell's viper toxin) RVV-X was used as an example of an X factor activator. RVV-X was purchased in the same manner as in Experimental Example 1, and the solution was dissolved according to the method specified by the manufacturer, and used at a volume of 20 μL per 180 μL of blood.

[0128] These reagents were placed in advance in a dielectric coagulometer measurement cartridge and heated to 37° C. before measurement in the dielectric coagulometer device.

[0129] [measurement] In the same manner as in Experimental Example 1, dielectric spectrum data was measured and recorded over time.

[0130] [analysis] As in Experimental Example 1, DBCM-CT was used for the analysis.

[0131] [result] 15 is a graph showing the DBCM-CT value versus blood concentration of rivaroxaban. As shown in FIG. 15, when a factor X activator was used, it was found that the electrical characteristics of the blood sample well reflected the blood concentration of rivaroxaban.

[0132] On the other hand, FIG. 16 shows the results of measuring the clotting time at each concentration of apixaban or rivaroxaban using TEG-6S (registered trademark) or ROTEM (registered trademark), which are blood viscoelasticity measuring devices. In both cases, the clotting time is shown as a percentage change when the clotting time when the concentration of apixaban or rivaroxaban is 0 is set to 1. The graph in FIG. 16 is quoted from the literature (Lapichino GE et al. Semin Thromb Hemost 2017;43:423-432). As shown in FIG. 16, when the factor X activator is not used, there is a large variation, and it is considered difficult to evaluate the efficacy of the anticoagulant.

[0133] These results show that when monitoring anticoagulants, by using a reagent that activates the precursor of the coagulation factor on which the anticoagulant acts, it is possible to accurately evaluate the efficacy of the anticoagulant from data on the change over time in the electrical properties of blood samples.

[0134] In addition, the present technology can also have the following configuration. (1) an inhibitor that inhibits factor Xa and / or thrombin; one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa; For the blood samples spiked with A blood coagulation system analyzer comprising an analysis unit that analyzes the effect of the inhibitor on the blood coagulation system by using data on changes over time in electrical properties measured at a specific frequency or frequency band. (2) The blood coagulation system analyzer according to (1), wherein the factor X activator is a factor X activating enzyme derived from snake venom. (3) The blood coagulation system analyzer according to claim 1 or 2, further comprising a reagent addition amount calculation unit that calculates the amount of the reagent to be added. (4) The blood coagulation system analyzer according to (3), wherein the reagent addition amount calculation unit calculates the amount of the reagent to be added based on the time-course data obtained from the blood sample of a healthy subject. (5) The blood coagulation system analyzer according to claim 4, wherein the reagent addition amount calculation unit calculates the amount of reagent to be added to be 0.5 to 2 when the amount of reagent to be added is 1 when the time-course data obtained from the blood sample of the healthy subject falls within a predetermined range. (6) A first time course data of electrical properties measured at a specific frequency or frequency band for a first blood sample to which one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa have been added; A second time-dependent change data of the electrical property measured at a particular frequency or frequency band for a second blood sample to which the reagent has not been added; and The blood coagulation system analyzer is provided with an analysis unit that utilizes the reagent to analyze the effects of coagulation factors upstream of the factor on which the reagent acts. (7) The blood coagulation system analyzer according to (6), wherein the analysis unit analyzes the influence of a coagulation factor upstream of a factor on which the reagent acts by correcting the second time-course data with the first time-course data. (8) A blood sample to which an anticoagulant and a reagent that activates a precursor of a coagulation factor on which the anticoagulant acts have been added is (9) A blood coagulation system analyzer comprising an analysis unit that analyzes the effect of the anticoagulant on the blood coagulation system by using data on changes over time in electrical characteristics measured at a specific frequency or frequency band. an inhibitor that inhibits factor Xa and / or thrombin; one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa; A measurement unit that measures electrical characteristics at a specific frequency or frequency band of a blood sample to which an analysis unit that analyzes the effect of the inhibitor on the blood coagulation system using data on the change over time of the measured electrical characteristics; A blood coagulation system measuring device comprising: (10) an inhibitor that inhibits factor Xa and / or thrombin; one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa; A measuring device for measuring electrical characteristics at a specific frequency or frequency band of a blood sample to which has been added; an analysis device that analyzes the effect of the inhibitor on the blood coagulation system using data on the change over time of the measured electrical characteristics; A blood coagulation system measuring system comprising: (11) a first blood sample to which one or more reagents selected from a factor X activator, a prothrombin activator, and factor Xa have been added; A measurement unit that measures electrical characteristics at a specific frequency or frequency band for a second blood sample to which the reagent has not been added; A first time-dependent change data of an electrical property measured from the first blood sample; and second time-dependent change data of the electrical property measured from the second blood sample; and an analysis unit that uses the above-mentioned reagent to analyze the effect of a coagulation factor upstream of the factor on which the reagent acts; A blood coagulation system measuring device comprising: (12) a first blood sample to which one or more reagents selected from a factor X activator, a prothrombin activator, and factor Xa have been added; a measuring device for measuring an electrical characteristic at a specific frequency or frequency band of a second blood sample to which the reagent has not been added; A first time-dependent change data of an electrical property measured from the first blood sample; and second time-dependent change data of the electrical property measured from the second blood sample; and an analysis device for analyzing the influence of a coagulation factor upstream of a factor on which the reagent acts, using the A blood coagulation system measuring system comprising: (13) an inhibitor that inhibits factor Xa and / or thrombin; one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa; For the blood samples spiked with A blood coagulation system analysis method that performs an analysis step of analyzing the effect of the inhibitor on the blood coagulation system by utilizing data on changes over time in electrical properties measured at a specific frequency or frequency band. (14) A first time course data of electrical properties measured at a specific frequency or frequency band for a first blood sample to which one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa have been added; A second time-dependent change data of the electrical property measured at a particular frequency or frequency band for a second blood sample to which the reagent has not been added; and The present invention relates to a blood coagulation system analysis method, and more particularly to a blood coagulation system analysis method. (15) an inhibitor that inhibits factor Xa and / or thrombin; one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa; A measuring step of measuring an electrical characteristic at a specific frequency or frequency band of the blood sample to which the an analysis step of analyzing the effect of the inhibitor on the blood coagulation system using data on the change over time of the measured electrical properties; A method for measuring the blood coagulation system. (16) a first blood sample to which one or more reagents selected from a factor X activator, a prothrombin activator, and factor Xa have been added; a measuring step of measuring electrical characteristics at a specific frequency or frequency band for a second blood sample to which the reagent has not been added; A first time-dependent change data of an electrical property measured from the first blood sample; and second time-dependent change data of the electrical property measured from the second blood sample; and an analysis step of analyzing the influence of a coagulation factor upstream of the factor on which the reagent acts, using the A method for measuring the blood coagulation system. [Explanation of symbols]

[0135] 1 Blood coagulation system analyzer 10. Blood coagulation analysis system 11 Signal acquisition section 12 Analysis Department 13 Reagent addition amount calculation section 14 Output section 15 Storage section 105 Storage device 16 Display 106 Display device 17,107 User Interface 2 Blood coagulation system measuring device 20 Blood coagulation system measurement system 21 Measuring part 201 Measuring Equipment 211 Blood sample holder 211a: Blood sample holder 211b: Container holding part 212a, 212b A pair of electrodes 213 Applicator 22 Drug Addition Section 23 Control Unit 203 Control device

Claims

1. an inhibitor that inhibits factor Xa and / or thrombin; one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa; For the blood samples spiked with A blood coagulation system analyzer comprising an analysis unit that analyzes the effect of the inhibitor on the blood coagulation system by using data on changes over time in electrical properties measured at a specific frequency or frequency band.

2. The blood coagulation system analyzer according to claim 1 , wherein the factor X activator is a factor X activating enzyme derived from snake venom.

3. The blood coagulation system analyzer according to claim 1 , further comprising a reagent addition amount calculation unit that calculates the amount of the reagent to be added.

4. 4. The blood coagulation system analyzer according to claim 3, wherein the reagent addition amount calculation unit calculates the amount of the reagent to be added based on the time-course change data obtained from the blood sample of a healthy subject.

5. 5. The blood coagulation system analyzer according to claim 4, wherein the reagent addition amount calculation unit calculates the amount of reagent to be added to be 0.5 to 2 when the amount of reagent to be added that brings the time-dependent change data obtained from the blood sample of the healthy subject into a predetermined range is set to 1.

6. A first time-dependent change data of an electrical characteristic measured at a specific frequency or frequency band for a first blood sample to which one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa have been added; A second time-dependent change data of the electrical property measured at a particular frequency or frequency band for a second blood sample to which the reagent has not been added; and The blood coagulation system analyzer is provided with an analysis unit that utilizes the reagent to analyze the effects of coagulation factors upstream of the coagulation factor on which the reagent acts.

7. 7. The blood coagulation system analyzer according to claim 6, wherein the analysis unit analyzes the influence of a coagulation factor upstream of a coagulation factor on which the reagent acts by correcting the second time-course data with the first time-course data.

8. A blood sample to which an anticoagulant and a reagent that activates a precursor of a coagulation factor on which the anticoagulant acts have been added is A blood coagulation system analyzer comprising an analysis unit that analyzes the effect of the anticoagulant on the blood coagulation system by using data on changes over time in electrical properties measured at a specific frequency or frequency band.

9. an inhibitor that inhibits factor Xa and / or thrombin; one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa; A measurement unit that measures electrical characteristics at a specific frequency or frequency band of a blood sample to which an analysis unit that analyzes the effect of the inhibitor on the blood coagulation system using data on the change over time of the measured electrical characteristics; A blood coagulation system measuring device comprising:

10. an inhibitor that inhibits factor Xa and / or thrombin; One or more reagents selected from factor X activator, prothrombin activator, and factor Xa Medicine and A measuring device for measuring electrical characteristics at a specific frequency or frequency band of a blood sample to which has been added; an analysis device that analyzes the effect of the inhibitor on the blood coagulation system using data on the change over time of the measured electrical characteristics; A blood coagulation system measuring system comprising:

11. a first blood sample to which one or more reagents selected from a factor X activator, a prothrombin activator, and factor Xa have been added; a measurement unit that measures an electrical characteristic at a specific frequency or frequency band for a second blood sample to which the reagent has not been added; a first time-varying electrical property data measured from the first blood sample; and second time-varying electrical property data measured from the second blood sample; and an analysis unit that uses the above-mentioned reagent to analyze the effect of a coagulation factor upstream of the coagulation factor on which the reagent acts; A blood coagulation system measuring device comprising:

12. a first blood sample to which one or more reagents selected from a factor X activator, a prothrombin activator, and factor Xa have been added; a measuring device for measuring an electrical characteristic at a specific frequency or frequency band of a second blood sample to which the reagent has not been added; a first time-varying electrical property data measured from the first blood sample; and second time-varying electrical property data measured from the second blood sample; and an analysis device for analyzing the influence of a coagulation factor upstream of the coagulation factor on which the reagent acts, using the A blood coagulation system measuring system comprising:

13. an inhibitor that inhibits factor Xa and / or thrombin; one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa; For the blood samples spiked with A blood coagulation system analysis method that performs an analysis step of analyzing the effect of the inhibitor on the blood coagulation system by utilizing data on changes over time in electrical properties measured at a specific frequency or frequency band.

14. A first time-dependent change data of an electrical characteristic measured at a specific frequency or frequency band for a first blood sample to which one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa have been added; A second time-dependent change data of the electrical property measured at a particular frequency or frequency band for a second blood sample to which the reagent has not been added; and The present invention relates to a blood coagulation system analysis method, and more particularly to a blood coagulation system analysis method.

15. an inhibitor that inhibits factor Xa and / or thrombin; one or more reagents selected from a factor X activator, a prothrombin activator, and a factor Xa; A measuring step of measuring an electrical characteristic at a specific frequency or frequency band of the blood sample to which the an analysis step of analyzing the effect of the inhibitor on the blood coagulation system using data on the change over time of the measured electrical properties; A method for measuring the blood coagulation system.

16. a first blood sample to which one or more reagents selected from a factor X activator, a prothrombin activator, and factor Xa have been added; a measuring step of measuring an electrical characteristic at a specific frequency or frequency band for a second blood sample to which the reagent has not been added; a first time-varying electrical property data measured from the first blood sample; and second time-varying electrical property data measured from the second blood sample; and an analysis step of analyzing the influence of a coagulation factor upstream of the coagulation factor on which the reagent acts, using the A method for measuring the blood coagulation system.

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