Method, device, system, and program for analyzing bleeding tendency of a subject

Abstract

To detect a patient with a tendency of an occurrence of serious bleeding based on a value acquired by a normal coagulation test.SOLUTION: An analysis method for analyzing bleeding tendency of a subject solves a problem by an analysis method by: acquiring a time series data set regarding coagulation reaction of a blood specimen collected from the subject; acquiring maximum speed arrival time up to when speed of coagulation reaction becomes maximum based on the acquired time series data set, maximum acceleration arrival time up to the time when the acceleration of the coagulation reaction is made maximum, and reference time in the coagulation reaction; acquiring a value based on the acquired maximum speed arrival time or the maximum acceleration arrival time and the reference time; and outputs acquired value or information regarding bleeding tendency based on the value.SELECTED DRAWING: Figure 14

Description

[Technical field]

[0001] The present invention relates to an analysis method, an analysis device, an analysis system, and an analysis program for analyzing a subject's bleeding tendency. [Background technology]

[0002] Patent Document 1 discloses a method for determining the severity of a patient with hemophilia A. In the method of Patent Document 1, a corrected linear curve is calculated from the coagulation reaction curve of the blood of a subject, and from the obtained corrected linear curve, the calculation target area value S is set to 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, and 90% of the maximum height value Vmax (100%), respectively, to extract 10 calculation target areas, and the flattening ratio vAB and the time rate vTB are calculated for the target area at each calculation target area value S. In addition, for blood (template specimen) of a patient with severe, moderate, and mild hemophilia A whose severity is determined based on the activity level of coagulation factor VIII (FVIII), the flattening ratio vAB and the time rate vTB are calculated from the corrected linear curve for the target area at each calculation target area value S. Then, a linear regression equation is calculated between the flattening ratio vAB and time rate vTB of the subject's blood and the flattening ratio vAB and time rate vTB of each template sample, and the template sample with the highest correlation is determined to determine the severity of the hemophilia A patient. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] International Publication No. 2020 / 158948 Summary of the Invention [Problem to be solved by the invention]

[0004] When a hemophilia A patient causes severe bleeding, it may leave aftereffects on the body, especially the joints, and the quality of life (QOL) may be significantly impaired. The severity of a hemophilia A patient is determined by measuring the activity level of FVIII. However, even a patient who is determined to be mild or moderate by measuring the activity level of FVIII may cause severe bleeding. Patent Document 1 discloses a method for determining the severity of a hemophilia A patient, but since the determination is based on the correlation with a template sample of a patient whose severity is determined based on the activity level of FVIII, it is not possible to detect a patient who is determined to be mild or moderate by measuring the activity level of FVIII and who subsequently causes severe bleeding. The present invention aims to detect a patient who causes severe bleeding, which could not be detected by the conventional measurement of the activity level of FVIII and the method of Patent Document 1. [Means for solving the problem]

[0005] The present invention relates to an analytical method for analyzing a bleeding tendency of a subject, which includes acquiring a time series dataset related to the coagulation reaction of a blood sample collected from the subject, acquiring a maximum velocity arrival time until the velocity of the coagulation reaction reaches a maximum or a maximum acceleration arrival time until the acceleration of the coagulation reaction reaches a maximum, and a reference time for the coagulation reaction based on the acquired time series dataset, acquiring a value based on the acquired maximum velocity arrival time or maximum acceleration arrival time and the reference time, and outputting the acquired value or information related to the bleeding tendency based on the value.

[0006] The present invention relates to an analytical device (100) that analyzes a bleeding tendency of a subject, the analytical device (100) including a control device (10), which acquires a time series data set relating to the coagulation reaction of a blood sample collected from the subject, acquires a maximum velocity arrival time until the velocity of the coagulation reaction reaches a maximum or a maximum acceleration arrival time until the acceleration of the coagulation reaction reaches a maximum, and a reference time for the coagulation reaction based on the acquired time series data set, acquires a value based on the acquired maximum velocity arrival time or maximum acceleration arrival time and the reference time, and outputs the acquired value or information relating to the bleeding tendency based on the value.

[0007] The present invention relates to an analysis program (132) for analyzing the bleeding tendency of a subject, which, when executed by a computer, causes the computer to execute the steps of: acquiring a time series data set regarding the coagulation reaction of a blood sample collected from the subject; acquiring a maximum velocity arrival time until the velocity of the coagulation reaction becomes maximum or a maximum acceleration arrival time until the acceleration of the coagulation reaction becomes maximum, and a reference time in the coagulation reaction based on the acquired time series data set; acquiring a value based on the acquired maximum velocity arrival time or maximum acceleration arrival time and the reference time; and outputting the acquired value or information regarding the bleeding tendency based on the value. Effect of the Invention

[0008] According to the present invention, it is possible to detect patients who develop severe bleeding, which could not be detected in the past. [Brief description of the drawings]

[0009] [Figure 1] FIG. 1 is a diagram showing the external appearance of an analysis system 1. [Diagram 2] FIG. 1 is a diagram showing a hardware configuration of an analysis system 1. [Diagram 3] FIG. 2 is a diagram showing information stored in an auxiliary storage device 13. [Figure 4] 2 is a diagram showing the configuration of a light irradiation unit of the measurement device 200. FIG. [Diagram 5] 1A is a diagram showing the configuration of the detection unit 22 when there is no cuvette 80. FIG. 1B is a diagram showing the configuration of the detection unit 22 when there is a cuvette 80. [Figure 6] FIG. 2 is a diagram showing the flow of measurement processing and analysis processing. [Figure 7] FIG. 13 is a diagram showing the flow of a measurement process. [Figure 8] FIG. 13 is a diagram showing the flow of an analysis process. [Figure 9] (A) An example of a time series data set, (B) a coagulation curve, and (C) a normalized coagulation curve. [Figure 10] (A) A diagram showing a first derivative coagulation curve, and (B) a diagram showing a second derivative coagulation curve. [Figure 11] FIG. 11 is a diagram showing the flow of a comparison process. [Figure 12] FIG. 13 is a diagram showing the flow of a modified example of the comparison process. [Figure 13] FIG. 1 shows 41 hemophilia A patients plotted on a two-dimensional scatter plot in which the vertical axis (Y-axis) indicates bleeding score value and the horizontal axis (X-axis) indicates FVIII activity. [Figure 14] (A) The clotting curve, first derivative clotting curve, and second derivative clotting curve of a healthy subject. (B) The clotting curve, first derivative clotting curve, and second derivative clotting curve of a subject with hemophilia A. [Figure 15] (A) Box plot with the vertical axis showing the value (S) obtained by formula 1, the box and whiskers on the left side showing the group of subjects with mild symptoms, and the box and whiskers on the right side showing the group of subjects with moderate symptoms or severe symptoms. (B) Box plot with the vertical axis showing the value (S) obtained by formula 2, the box and whiskers on the left side showing the group of subjects with mild symptoms or moderate symptoms, and the box and whiskers on the right side showing the group of subjects with severe symptoms. (C) Box plot with the vertical axis showing the value (S) obtained by formula 3, the box and whiskers on the left side showing the group of subjects with mild symptoms or moderate symptoms, and the box and whiskers on the right side showing the group of subjects with severe symptoms. [Figure 16](A) A box plot in which the vertical axis shows the value (S) obtained by Formula 2, the boxes and whiskers on the left side show the group of subjects with mild or moderate symptoms, and the boxes and whiskers on the right side show the group of subjects with severe symptoms. (B) A box plot in which the vertical axis shows the value (S) obtained by Formula 3, the boxes and whiskers on the left side show the group of subjects with mild or moderate symptoms, and the boxes and whiskers on the right side show the group of subjects with severe symptoms. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] 1.Analysis System 1 The analysis system 1 will be described with reference to FIGS.

[0011] As shown in FIG. 1, the analysis system 1 includes an analysis device 100 that analyzes a subject's bleeding tendency, and a measurement device 200 that includes a detection unit that irradiates light onto a blood sample that has initiated a coagulation reaction and acquires optical information.

[0012] The blood sample to be analyzed by the analysis system 1 is blood collected from a subject, or a sample obtained by processing the collected blood. Examples of blood samples include whole blood and plasma. It is preferable to collect blood using an anticoagulant other than a heparin preparation when collecting blood. Citrate, for example, a sodium citrate solution, can be used as the anticoagulant. For example, plasma separated from a sample in which a 3.1% to 3.3% (weight / volume) trisodium citrate solution is used as the anticoagulant and the collected blood is mixed at a volume ratio of about 1:8.5 to 1:9.5 can be used as the blood sample.

[0013] The subject from which blood is collected is, but is not limited to, a subject suspected of having a bleeding tendency. The subject suspected of having a bleeding tendency is, for example, a patient with low factor VIII activity. The cause of the decrease in factor VIII activity includes functional factors and quantitative factors of factor VIII. The subject suspected of having a bleeding tendency is, for example, a patient with hemophilia A.

[0014] 1-1.Analysis device 100 FIG. 2 is a diagram showing the hardware configuration of the analysis system 1. As shown in FIG. Analysis device 100 is communicatively connected to measurement device 200. Analysis device 100 is also connected to a media drive 98. Analysis device 100 is also connected to electronic medical record system 1000 via network 99. Network 99 is, for example, a local area network (LAN), but may also be the Internet.

[0015] The analysis device 100 includes a control device 10, an input device 16, and an output device 17. The control device 10 includes a CPU (Central Processing Unit) 11 for performing data processing, a main storage device 12 used as a working area for data processing, an auxiliary storage device 13 for storing information transferred to the main storage device 12, a bus 14 for transmitting data between each device, and an interface (I / F) 15 for inputting and outputting data to and from an external device. The input device 16, the output device 17, a media drive 98, a network 99, and a measurement device 200 are connected to the interface (I / F) 15. The interface (I / F) 15 is, for example, a USB, IEEE1394, or Ethernet. The auxiliary storage device 13 is, for example, a solid state drive or a hard disk drive. The input device 16 is, for example, a keyboard or a mouse, and the output device 17 is, for example, a liquid crystal display or an organic EL display. Information regarding the bleeding tendency of the subject is output to the output device 17. 3 is a diagram showing information stored in the auxiliary storage device 13. The auxiliary storage device 13 stores a measurement program 131 for the measurement device 200 to perform blood coagulation measurement, an analysis program 132 for analyzing the subject's bleeding tendency, a function database (DB) DB1 that stores mathematical expressions used in the analysis, and a reference value database (DB) DB2 that stores reference values.

[0016] 1-2. Configuration of the measuring device Returning to Fig. 2, the hardware configuration of the measuring device 200 will be described. The measuring device 200 includes a sample preparation unit 20 that prepares a sample so that the blood sample starts a coagulation reaction, a light irradiation unit 27, a detection unit 22, and an interface (I / F) 25. The interface (I / F) 15 is, for example, USB, IEEE1394, or Ethernet.

[0017] FIG. 4 shows the configuration of the light irradiation unit 27. The light irradiation unit 27 includes five light sources 321, 322, 323, 324, and 325, five optical fiber units 330a, 330b, 330c, 330d, and 330e provided corresponding to the five light sources 321 to 325, and one holding member 340 for holding the light sources 321 to 325 and the incident ends 331 of the optical fiber units 330a to 330e. The light sources 321 to 325, the optical fiber units 330a to 330e, and the holding member 340 are housed in a metal housing 310. The light sources 321 to 325 are all composed of LEDs, and the first light source 321 generates light of 660 nm. The second light source 322 generates light of 405 nm. The third light source 323 generates light of 800 nm. The fourth light source 324 generates light of 340 nm. The fifth light source 325 emits light of 575 nm. The light sources 321 to 325 emit light under the control of the control device 10 of the measurement device 200.

[0018] Each of the optical fiber parts 330a to 330e is configured as a cable in which optical fiber strands each having one core are bundled. The optical fiber parts 330a to 330e are bundled together at the intermediate part 333, and the bundled optical fiber parts 330a to 330e are divided into two bundles, and the emission ends 332 of the bundles are held by the outlet 311 provided in the housing 310. The outlet 311 also holds the incident end 352 of the optical fiber 21 that connects the light irradiation part 27 and the detection part 22. Between the emission end 332 of the optical fiber parts 330a to 330e and the incident end 352 of the optical fiber 21, a homogenizing member 350 for homogenizing the intensity distribution of the light emitted from the emission end 332 is provided. The homogenizing member 350 is a member that causes multiple reflections of the light incident from the incident surface 351 inside, and is, for example, a polygonal rod homogenizer.

[0019] 5(A) and (B) show the configuration of the detection unit 22. The detection unit 22 is provided in each optical fiber 21 connected to the light irradiation unit 27. However, the connection configuration between the optical fiber 21 and the detection unit 22 and the hardware configuration of the detection unit 22 are the same for all the detection units 22, so one detection unit 22 will be described here. FIG. 5(A) is a diagram showing the detection unit 22 in a state where it does not hold a cuvette 80 for mixing a coagulation time measurement reagent and a blood sample. The detection unit 22 has a hole 22b formed therein into which the other end 353 of the optical fiber 21, whose entrance end 352 (FIG. 4) is held at the outlet 311 of the light irradiation unit 27, is inserted. Furthermore, the detection unit 22 has a communication hole 22c formed therein, which communicates the hole 22b with the holding unit 22a for holding the cuvette 80.

[0020] The diameter of the hole 22b is larger than the diameter of the communication hole 22c. A lens 22d that collects light from the optical fiber 21 is disposed at the end of the hole 22b. Furthermore, a hole 22f is formed in the inner wall surface of the holding part 22a at a position facing the communication hole 22c. A light receiving part 22g is disposed at the back of this hole 22f. The light receiving part 22g is, for example, a photodiode, and outputs an electrical signal according to the amount of light received. The light transmitted through the lens 22d is incident on the light receiving surface of the light receiving part 22g via the communication hole 22c, the holding part 22a, and the hole 22f. The optical fiber 21 is fixed by the leaf spring 22e with the end 353 inserted into the hole 22b.

[0021] FIG. 5(B) is a diagram showing the detection unit 22 in a state where the cuvette 80 is held. When the cuvette 80 is held in the holding unit 22a, the light collected by the lens 22d passes through the cuvette 80 and the sample contained in the cuvette 80 and enters the light receiving unit 22g. When the blood coagulation reaction in the sample progresses, the turbidity of the sample increases. Accordingly, the amount of light passing through the sample (amount of transmitted light) decreases, and the level of the electrical signal output from the light receiving unit 22g decreases. The electrical signal output from the light receiving unit 22g is converted into a digital signal by the A / D converter 22h and transmitted to the analysis device 100 via the communication I / F 25 (see FIG. 1). The signal output from the light receiving unit 22g and converted by the A / D converter 22h is optical information reflecting the amount of transmitted light.

[0022] It should be noted that the measuring device 200 may be the device described in US Pat. No. 10,048,249, the specification of which is incorporated herein by reference.

[0023] 1-3. Measurement and analysis 6 shows a flow of measurement and analysis processing by control device 10 of analysis device 100. In step S1, control device 10 executes measurement processing based on measurement program 131. Next, in step S2, control device 10 executes analysis processing based on analysis program 132.

[0024] 1-4. Processing of measurement programs FIG. 7 shows the flow of the measurement process executed by the control device 10 based on the measurement program 131. The control device 10 receives a measurement start request input by the operator from the input device 16, and in step S11, acquires information on blood coagulation parameters (test items) ordered for each blood sample based on the information input by the operator from the input device 16. The control device 10 may acquire information on blood coagulation parameters via the network 99, for example, from an electronic medical record system 1000 of a medical facility.

[0025] Hereinafter, a case where the blood coagulation parameter (test item) acquired in step S11 is activated partial thromboplastin time (hereinafter sometimes abbreviated as "APTT") will be described. When measuring APTT, a coagulation time measuring reagent and a coagulation activation reagent are used. As the coagulation time measuring reagent, an APTT measuring test reagent containing an activator such as silica, ellagic acid, celite, etc. and an animal-derived, plant-derived, or artificially synthesized phospholipid, etc., can be used. As the APTT measuring test reagent, Thrombocheck (registered trademark) APTT series manufactured by Sysmex Corporation, Coagpia (registered trademark) APTT-N manufactured by Sekisui Medical Co., Ltd., and Dataphi-APTT manufactured by Siemens Healthcare Diagnostics Products GmbH can be exemplified. The coagulation activation reagent is a reagent capable of supplying calcium ions, for example, a 20 mM calcium chloride solution.

[0026] The control device 10 controls the sample preparation unit 20 of the measurement device 200 to dispense a blood sample into the cuvette 80 in step S12.

[0027] In step S13, the control device 10 controls the sample preparation unit 20 of the measurement device 200 to prepare a measurement sample by dispensing a coagulation time measuring reagent corresponding to the blood coagulation parameter acquired in step S11 into the cuvette 80 into which the blood sample has been dispensed. In addition, in step S13, the control device 10 controls the sample preparation unit 20 of the measurement device 200 to dispense a coagulation activation reagent into the blood sample into which the coagulation time measuring reagent has been dispensed.

[0028] In step S14, the control device 10 controls the light irradiating unit 27 to start irradiating light to the cuvette 80 into which the coagulation activation reagent was dispensed in step S13. In this example, the control device 10 controls the light irradiating unit 27 to irradiate light only from the first light source 321. The control device 10 continuously receives a signal (digital data) output from the light receiving unit 22g according to the intensity of light received through the cuvette 80 and the measurement sample from the time when the coagulation activation reagent was dispensed into the cuvette 80 until a predetermined measurement time (for example, 120 seconds to 180 seconds) is reached. The control device 10 stores the continuously received digital data in the auxiliary storage device 13 as a time series data set related to the coagulation reaction of the blood sample. The time interval at which the control device 10 receives the digital data is, for example, 0.1 seconds to 0.5 seconds.

[0029] 1-5. Processing of analysis programs FIG. 8 shows a flow of the analysis process for analyzing the bleeding tendency of a subject, which is executed by the control device 10 based on the analysis program 132. The control device 10 starts the analysis process by receiving a measurement start request input by the operator from the input device 16. The control device 10 may start the analysis process by using the end of step S14 shown in FIG. 7 as a trigger. The control device 10 may also measure the FVIII activity level in advance in step S1 shown in FIG. 6, compare the FVIII activity level with a predetermined value (for example, 10%), and start the analysis process when the FVIII activity level is smaller than the predetermined value. The predetermined value is not particularly limited, but is preferably an activity level that is diagnosed as mild or moderate in the severity assessment of hemophilia A patients. When the FVIII activity level is smaller than the predetermined value, there is a high possibility that the subject will later develop severe bleeding, compared with when it is larger than the predetermined value. Therefore, by executing the analysis process on subjects whose FVIII activity level is smaller than the predetermined value, patients who will develop severe bleeding can be efficiently detected.

[0030] In step S21, the control device 10 acquires a time series data set relating to the coagulation reaction of the blood sample collected from the subject. Specifically, the control device 10 reads out the time series data set stored in the auxiliary storage device 13 in step S14.

[0031] The time series data set will be described in more detail with reference to Figs. 9(A) and (B). In this example, the time series data set is a set of two-dimensional array data consisting of the intensity of light (hereinafter referred to as "transmitted light intensity") transmitted through a measurement sample when the measurement sample is irradiated with light, and the time when the transmitted light is detected. Fig. 9(A) is a diagram showing an example of a time series data set. In the time series data set of Fig. 9(A), the upper row indicates the transmitted light intensity, and the lower row indicates the time when each transmitted light intensity was acquired (time (t I 9(B) shows the blood coagulation curve when the time series array data set is plotted on a two-dimensional graph in which the vertical axis (Y-axis) indicates the transmitted light intensity and the horizontal axis (X-axis) indicates the measurement time (seconds: sec) during which the transmitted light intensity was monitored.

[0032] Next, in S22 shown in FIG. 8, the control device 10 normalizes the blood coagulation curve indicated by the time series data set acquired in step S21, differentiates the obtained blood coagulation curve to obtain a first differential coagulation curve, and differentiates the first differential coagulation curve to obtain a second differential coagulation curve.

[0033] Fig. 9(C) shows a blood coagulation curve obtained by normalizing the blood coagulation curve shown in Fig. 9(B). As shown in Fig. 9(C), in the blood coagulation curve obtained by normalization, each transmitted light intensity included in the time series data set is expressed as a relative value so that the vertical axis is represented between 0% (L1: baseline) and 100% (L2).

[0034] Figure 10(A) shows a first differential coagulation curve obtained by differentiating the blood coagulation curve shown in Figure 9(C). Figure 10(B) shows a second differential coagulation curve obtained by differentiating the first differential coagulation curve shown in Figure 10(A).

[0035] In step S23 shown in FIG. 8, the control device 10 determines a reference time (t c ) is obtained. c ) is, for example, the time it takes for a blood sample to reach a predetermined coagulation state after the start of clotting.

[0036] Reference time (t c The acquisition of time (t I ) is the time when the coagulation activation reagent is added to the blood sample. I At time (t), the fibrinogen in the blood sample has not been converted to fibrin, and fibrin precipitation does not occur, so the transmitted light intensity is high. As the coagulation reaction progresses and fibrin begins to precipitate, the precipitated fibrin blocks the light, and the transmitted light intensity begins to decrease. This point is called the time (t II ), which is the coagulation reaction start time. The control device 10 obtains the time at which the transmitted light intensity starts to decrease in the blood coagulation curve by calculation, and calculates the time (t II ) and the control device 10 determines the time (t II ) the change in transmitted light intensity is set to 0%.

[0037] Thereafter, as the reaction progresses and fibrin precipitation progresses, the transmitted light intensity further decreases. When most of the fibrinogen in the blood sample is converted to fibrin, the reaction converges and the blood coagulation curve plateaus. The control device 10 calculates the time when the blood coagulation curve begins to plateau and determines the time (t III ) time (t III ) is the time when the coagulation reaction ends. The control device 10 also determines the time (t III ) is set to 100%. Next, the control device 10 sets the time when the change in the transmitted light intensity is 50% (the time at the intersection of the line L3 and the blood coagulation curve) as the reference time (t c ) is obtained as the reference time (t cIn this embodiment, the reference time (t c ) is, for example, the time (t II ) may be a certain period of time counted from the Note that normalization of the blood coagulation reaction curve can be omitted. In that case, the control device 10 obtains a first derivative coagulation curve by differentiating the blood coagulation curve shown in FIG. 9(B). The control device 10 also obtains a reference time (t c ) to get the

[0038] In step S23 shown in FIG. 8, the control device 10 calculates the coagulation reaction start time (t II The time from the time when the coagulation reaction reaches its maximum (t min1 The control device 10 calculates the maximum speed arrival time (t min1 ) based on the second derivative coagulation curve shown in FIG. 10(B), the coagulation reaction initiation time (t II ) until the acceleration of the coagulation reaction becomes 0 (t a0 ) may be obtained by calculation.

[0039] In step S23 shown in FIG. 8, the control device 10 calculates the coagulation reaction start time (t II The time from when the acceleration of the coagulation reaction reaches its maximum to when the acceleration reaches its maximum (t min2 ) is obtained by calculation.

[0040] In step S23 shown in FIG. 8, the control device 10 calculates, based on the first derivative coagulation curve shown in FIG. 10(A) acquired in step S22, the absolute value |min1| of the maximum value min1 of the first derivative value as the maximum coagulation reaction rate.

[0041] In step S23 shown in FIG. 8, based on the second derivative coagulation curve shown in FIG. 10(B) obtained in step S22, the control device 10 calculates, by calculation, |min2|, which is the absolute value of the maximum value min2 of the second derivative, as the maximum coagulation reaction acceleration, and calculates, by calculation, |max2|, which is the absolute value of the minimum value max2 of the second derivative, as the maximum coagulation reaction deceleration (i.e., the minimum value of the coagulation reaction acceleration).

[0042] In step S24 shown in Fig. 8, the control device 10 acquires a value (S) based on the value acquired in step S23. This value (S) reflects how quickly the blood coagulation reaction progresses after the start of the blood coagulation reaction. The value (S) also reflects that in the case of blood from a subject with a low bleeding tendency, the rate of the blood coagulation reaction is maximized at approximately the middle point of the blood coagulation reaction, and in the case of blood from a subject with a high bleeding tendency, the rate of the blood coagulation reaction is maximized at a point earlier than the middle point of the blood coagulation reaction.

[0043] The formula for calculating the value (S) is stored in a function database DB1. The value (S) can be expressed by the following formula: Value(S)=(t c -t min1 )×(|min1|) (Formula 1) ("-" indicates subtraction. "×" indicates multiplication.)

[0044] 8, the control device 10 compares the value (S) acquired in step S24 with a predetermined reference value. The predetermined reference value is a reference value for determining the level of the bleeding tendency of the subject, and is set in advance and stored in the reference value database DB2 of the auxiliary storage device 13.

[0045] The control device 10 outputs a result in step S26 shown in Fig. 8 based on the comparison between the value and the reference value performed in step S25. The result may include information regarding bleeding tendency.

[0046] The information on the bleeding tendency includes a label indicating the degree of the bleeding tendency. The label indicating the degree of the bleeding tendency may indicate the degree of the bleeding tendency by a numerical value or a symbol, or may indicate the degree of the bleeding tendency by a sentence or a word. The label indicating the degree of the bleeding tendency includes a first label indicating a low bleeding tendency and a second label indicating a medium or high bleeding tendency. The reference value used in the comparison in step S25 is a first reference value for determining whether the subject has a low bleeding tendency or a medium or high bleeding tendency.

[0047] The first and second labels are associated with the reference values ​​and stored in advance in the reference value database DB2.

[0048] The first and second reference values ​​can be set to correspond to bleeding score values ​​according to, for example, the International Society on Thrombosis and Haemostasis Bleeding Assessment Tool (ISTH-BAT) provided by the Japanese Society on Thrombosis and Hemostasis. That is, a value (S) is obtained for blood samples of multiple subjects whose ISTH-BAT bleeding score values ​​are known, and the first and second reference values ​​are set so that, for example, a first label is output for subjects whose ISTH-BAT bleeding score value is 10 or less, and a second label is output for subjects whose ISTH-BAT bleeding score value is greater than 10.

[0049] 8, the control device 10 may output the value (S) acquired in step S24 as a result in step S26. Even if the control device 10 has performed step S25, the control device 10 may output the value (S) acquired in step S24 as a result in step S26. The control device 10 may output the value (S) acquired in step S24 and information relating to bleeding tendency based on the comparison result in step S25 in step S26.

[0050] The comparison process in step S25 will be described with reference to FIG. In step S51, the value (S) is compared with the first reference value, and if the value (S) is greater (in the case of "YES"), the control device 10 proceeds to step S53 and selects a second label stored in the reference value database DB2 indicating that the bleeding tendency is moderate or high. The control device 10 proceeds to step S26 shown in Fig. 8 and outputs the second label selected in step S53 to the output device 17 as information indicating that the bleeding tendency of the subject is moderate or high.

[0051] In step S51, if the value (S) is equal to or less than the first reference value (in the case of "NO"), the control device 10 proceeds to step S55 and selects a first label indicating a low bleeding tendency stored in the reference value database DB2. The control device 10 proceeds to step S26 shown in Fig. 8 and outputs the first label selected in step S55 to the output device 17 as information indicating that the subject has a low bleeding tendency.

[0052] 1-6. Modification of analysis system 1 In the above example, in step S11 (FIG. 7), APTT is obtained as a blood coagulation parameter (test item), but the blood coagulation parameter (test item) may also be, for example, prothrombin time (hereinafter sometimes abbreviated as "PT"), thrombotest, fibrinogen, factor VIII activity, factor II activity, factor V activity, factor VII activity, factor IX activity, factor X activity, factor XI activity, factor XII activity, or whole blood clotting time.

[0053] When PT is obtained as a blood coagulation parameter (test item), a PT measurement test reagent containing thrombin can be used as a coagulation time measurement reagent. Examples of PT measurement test reagents include the Thrombocheck (registered trademark) PT series manufactured by Sysmex Corporation and the Coagpia (registered trademark) PT series manufactured by Sekisui Medical Co., Ltd. PT measurement test reagents generally contain calcium ions necessary for coagulation activation.

[0054] In the above example, in step S24, Equation 1 is used as the equation for obtaining the value (S). However, the value (S) may be obtained, for example, by calculating the reference time (t c ) to reach maximum speed (t min1 ) or the reference time (t c ) to maximum acceleration arrival time (t min2 ) may be subtracted.

[0055] Also, the value (S) is the reference time (t c ) to reach maximum speed (t min1 ) or the reference time (t c ) to maximum acceleration arrival time (t min2 ) and a coefficient. The coefficient may be, for example, |min1|, |min2|, or a coefficient based on |min2| and |max2|. For example, the coefficient based on |min2| and |max2| may be the difference between the two values ​​(|min2|-|max2|, or |max2|-|min2|) ("-" indicates subtraction), or the division value of the two values ​​(|min2| / |max2|, or |max2| / |min2|) (" / " indicates division). By using the coefficient, the value (S) can more strongly reflect how fast the blood coagulation reaction progressed after the start of the coagulation reaction, and the detection sensitivity of subjects with a high tendency to bleed can be improved.

[0056] The value (S) may be, for example, a value represented by the following formula: Value(S)=(t c -t min1 )×(|min2|) (Formula 2) Value(S)=(t c -t min1 ) × (|min2|-|max2|) (Formula 3) Value(S)=(t c -t min1 ) × (|max2|-|min2|) (Formula 3') Value(S)=(t c -t min1 ) × (|min2| / |max2|) (Formula 4) Value(S)=(t c -t min1)×(|max2| / |min2|) (Formula 4') Value(S)=(t c -t min1 ) (Equation 5) Value(S)=(t c -t min2 )×(|min1|) (Formula 6) Value(S)=(t c -t min2 )×(|min2|) (Formula 7) Value(S)=(t c -t min2 ) × (|min2|-|max2|) (Formula 8) Value(S)=(t c -t min2 ) × (|max2|-|min2|) (Formula 8') Value(S)=(t c -t min2 ) × (|min2| / |max2|) (Formula 9) Value(S)=(t c -t min2 ) × (|max2| / |min2|) (Formula 9'), or Value(S)=(t c -t min2 ) (Equation 10) ("-" indicates subtraction, "x" indicates multiplication, and " / " indicates division.)

[0057] FIG. 12 shows a modification of the comparison process shown in FIG. In this case, the label indicating the degree of bleeding tendency includes a third label indicating a low or moderate bleeding tendency, and a fourth label indicating a high bleeding tendency. The reference value is a second reference value for determining whether the subject has a low or moderate bleeding tendency, or a high bleeding tendency. The first and second reference values ​​are set so that the third label is output for a subject with an ISTH-BAT bleeding score of 15 or less, and the fourth label is output for a subject with an ISTH-BAT bleeding score of more than 15.

[0058] In step S61, the value (S) is compared with the second reference value, and if the value (S) is greater (in the case of "YES"), the control device 10 proceeds to step S63 and selects a fourth label that indicates a high bleeding tendency stored in the reference value database DB2. The control device 10 proceeds to step S26 shown in Fig. 8 and outputs the fourth label selected in step S63 to the output device 17 as information that indicates that the subject has a high bleeding tendency.

[0059] In step S61, if the value (S) is equal to or less than the second reference value (in the case of "NO"), the control device 10 proceeds to step S65 and selects a third label stored in the reference value database DB2, which indicates that the bleeding tendency is low or moderate. The control device 10 proceeds to step S26 shown in Fig. 8 and outputs the third label selected in step S65 to the output device 17 as information indicating that the bleeding tendency of the subject is low or moderate.

[0060] In the above example, one of the first and second reference values ​​is used, but both reference values ​​may be used to determine whether the subject has a low bleeding tendency, a medium bleeding tendency, or a high bleeding tendency.

[0061] Furthermore, in the above example, the light receiving unit 22g of the detection unit 22 detects light (transmitted light) that has passed through the cuvette 80 and the sample contained in the cuvette 80, but the detection unit 22 may also be configured to receive light (scattered light) scattered by the sample contained in the cuvette 80, and the control device 10 may analyze the blood coagulation reaction curve based on changes in the scattered light intensity.

[0062] 2.Verification of effectiveness FIG. 13 shows a diagram in which ISTH-BAT bleeding score values ​​were calculated for 41 subjects who were determined to have mild or moderate hemophilia as a result of diagnosing the severity of hemophilia based on FVIII activity measurement, and each subject was plotted on a two-dimensional scatter plot in which the vertical axis (Y axis) indicates the bleeding score value and the horizontal axis (X axis) indicates FVIII activity. Box P shown in FIG. 13 is an area to which a group of subjects whose ISTH-BAT bleeding score value is greater than 15 and whose FVIII activity is less than 10% belong. In the severity diagnosis of hemophilia based on FVIII activity measurement, if the FVIII activity is 1% or more and less than 5%, it is determined to be moderate, and if the FVIII activity is 5% or more and less than 40%, it is determined to be mild. Therefore, although subjects belonging to box P have ISTH-BAT bleeding score values ​​greater than 15 and may cause severe bleeding, they are determined to have mild or moderate hemophilia in the diagnosis based on FVIII activity measurement.

[0063] Figure 14(A) shows, from top to bottom, the blood coagulation curve, the first derivative coagulation curve, and the second derivative coagulation curve of a healthy subject without hemophilia. Figure 14(B) shows, from top to bottom, the blood coagulation curve, the first derivative coagulation curve, and the second derivative coagulation curve of a subject with a bleeding score of 25 on the ISTH-BAT and a high tendency to bleed (a subject belonging to box P shown in Figure 13).

[0064] As shown in FIG. 14(A), in the case of a healthy subject, the reference time (t c ) and time to reach maximum speed (t min1 ) are almost the same time. Therefore, the value (S) is close to 0, so in the comparison process shown in FIG. 11 and FIG. 12, the first label indicating a low bleeding tendency or the third label indicating a low or medium bleeding tendency is selected. On the other hand, as shown in FIG. 14(B), in the case of a subject with a high bleeding tendency, the reference time (t c ) and time to reach maximum speed (t min1 ) is greater than the difference in healthy subjects, the second label indicating a medium or high bleeding tendency or the fourth label indicating a high bleeding tendency is selected in the comparison process shown in Figures 11 and 12.

[0065] Similarly, the reference time (t c ) and time to reach maximum acceleration (t min2 ) is also greater in subjects with a high bleeding tendency than in healthy subjects, so for subjects with a high bleeding tendency, a second label indicating a medium or high bleeding tendency or a fourth label indicating a high bleeding tendency is selected.

[0066] The effect of the present invention was verified by measuring APTT and obtaining values ​​(S) using the plasma of 41 subjects with hemophilia A used in creating the two-dimensional scatter plot of FIG. 13. The results are shown in FIG. 15(A) to (C). FIG. 15(A) to (C) are box plots in which the vertical axis (Y axis) indicates the value (S). In creating the box plot of FIG. 15(A), the 41 subjects were divided into a subject group with an ISTH-BAT bleeding score of 10 or less (a mild subject group) and a subject group with an ISTH-BAT bleeding score of more than 10 (a moderate or severe subject group). FIG. 15(A) is a box plot in which the vertical axis indicates the value (S) obtained by Formula 1, the box and whiskers on the left side indicate the mild subject group, and the box and whiskers on the right side indicate the moderate or severe subject group.

[0067] As shown in FIG. 15(A), by using the value (S), it is possible to distinguish between a group of subjects with mild symptoms, who have an ISTH-BAT bleeding score value of 10 or less, and a group of subjects with moderate or severe symptoms, who have an ISTH-BAT bleeding score value of more than 10, and the value (S) and the ISTH-BAT bleeding score value have a high correlation.

[0068] Similarly, in creating the box plots of Figures 15(B) and (C), 41 subjects were divided into a subject group with ISTH-BAT bleeding score of 15 or less (mild or moderate subject group) and a subject group with ISTH-BAT bleeding score of more than 15 (severe subject group). Figure 15(B) is a box plot in which the vertical axis indicates the value (S) obtained by Formula 2, the box and whiskers on the left side indicate the mild or moderate subject group, and the box and whiskers on the right side indicate the severe subject group. As shown in Figure 15(B), by using the value (S), it is possible to distinguish between the mild or moderate subject group with ISTH-BAT bleeding score of 15 or less and the severe subject group with ISTH-BAT bleeding score of more than 15, and the value (S) and the ISTH-BAT bleeding score value have a high correlation.

[0069] Fig. 15(C) is a box plot in which the vertical axis indicates the value (S) obtained by Formula 3, the box and whiskers on the left side indicate the mild or moderate subject group, and the box and whiskers on the right side indicate the severe subject group. As shown in Fig. 15(C), by using the value (S), it is possible to distinguish between the mild or moderate subject group whose ISTH-BAT bleeding score value is 15 or less and the severe subject group whose ISTH-BAT bleeding score value is greater than 15, and the value (S) and the ISTH-BAT bleeding score value have a high correlation.

[0070] As described above, the value (S) is highly correlated with the ISTH-BAT bleeding score value, and by using the value (S), it is possible to detect patients who are judged to have mild or moderate bleeding in the test based on FVIII measurement but who belong to the frame P ( FIG. 13 ) having a high bleeding score value, i.e., patients who cause severe bleeding that could not be detected by measuring the FVIII activity level.

[0071] Next, in preparing the box plots of Figures 16(A) and (B), subjects with FVIII activity levels of less than 10% were extracted from the 41 subjects with hemophilia A used in preparing the two-dimensional scatter plot of Figure 13, and the extracted subjects were divided into a subject group with an ISTH-BAT bleeding score value of 15 or less (a mild or moderate subject group) and a subject group with an ISTH-BAT bleeding score value of more than 15 (a severe subject group). Figure 16(A) is a box plot in which the vertical axis indicates the value (S) obtained by Formula 2, the box and whiskers on the left side indicate the mild or moderate subject group, and the box and whiskers on the right side indicate the severe subject group. As shown in FIG. 16(A), by extracting subjects with FVIII activity levels below 10%, the discrimination performance between the mild or moderate subject group and the severe subject group was improved compared to the case where all subjects were included (see FIG. 15(B)). Similarly, FIG. 16(B) is a box plot in which the vertical axis indicates the value (S) obtained by Equation 3, the box and whiskers on the left side indicate the mild or moderate subject group, and the box and whiskers on the right side indicate the severe subject group. As shown in FIG. 16(B), by extracting subjects with FVIII activity levels below 10%, the discrimination performance between the mild or moderate subject group and the severe subject group was improved compared to the case where all subjects were included (see FIG. 15(C)). Therefore, by extracting subjects with FVIII activity levels below a predetermined value, patients who cause severe bleeding that could not be detected by FVIII activity level measurement can be more efficiently detected. [Explanation of symbols]

[0072] 100 Analyzer 10 Control device 22 Detection unit 132 Analysis Program

Claims

1. An analysis method for analyzing a bleeding tendency of a subject, comprising: obtaining a time series data set relating to a coagulation reaction of a blood sample taken from the subject; Based on the acquired time series data set, a maximum velocity arrival time until the velocity of the coagulation reaction becomes maximum, or a maximum acceleration arrival time until the acceleration of the coagulation reaction becomes maximum, and a reference time in the coagulation reaction are acquired; Obtaining a value based on the acquired maximum velocity arrival time or the acquired maximum acceleration arrival time and the reference time; outputting the acquired value or information regarding bleeding tendency based on the value; The value indicates the degree of bleeding tendency. The analysis method.

2. In obtaining the value, The value is obtained based on the maximum velocity arrival time or the maximum acceleration arrival time, the reference time, and a coefficient. The analysis method according to claim 1 .

3. The coefficient includes a maximum value of the coagulation reaction rate, a maximum value of the coagulation reaction acceleration, or a difference between the maximum value and the minimum value of the coagulation reaction acceleration. The analysis method according to claim 2.

4. The reference time is the time from the start of the coagulation reaction until the blood sample reaches a predetermined coagulation state. The analysis method according to any one of claims 1 to 3.

5. The information regarding the bleeding tendency based on the value includes a label indicating the degree of the bleeding tendency. The analysis method according to any one of claims 1 to 4.

6. The label includes a first label indicating a low or moderate bleeding tendency, and a second label indicating a high bleeding tendency. The analysis method according to claim 5.

7. The labels include a third label indicating a low bleeding tendency, and a fourth label indicating a medium or high bleeding tendency. The analysis method according to claim 5 or 6.

8. In obtaining the value, obtaining the value based on the maximum speed reaching time and the reference time; The analysis method according to any one of claims 1 to 7.

9. The value is obtained by any one of the following formulas: The analysis method according to any one of claims 1 to 8, Value (S) = (t c -t min1 ) × (|min1|) (Formula 1) Value (S) = (t c -t min1 ) × (|min2|) (Formula 2) Value (S) = (t c -t min1 ) × (|min2|-|max2|) (Formula 3) (t c denotes the reference time, and t min1 indicates the maximum velocity arrival time, |min1| indicates the absolute value of the coagulation reaction velocity at the maximum velocity arrival time, |min2| indicates the absolute value of the coagulation reaction acceleration at the maximum acceleration arrival time, and |max2| indicates the absolute value of the maximum coagulation reaction deceleration at which the acceleration of the coagulation reaction is minimum. In the formulas, "-" indicates subtraction and "x" indicates multiplication.

10. Furthermore, the activity level of coagulation factor VIII is measured, When the measured activity level of coagulation factor VIII is smaller than a predetermined value, the obtained value or information on bleeding tendency based on the value is output. The analysis method according to any one of claims 1 to 9.

11. The method according to claim 1 , wherein the coagulation reaction of the blood sample is measured by activated partial thromboplastin time.

12. The method of claim 1 , wherein the subject has hemophilia A.

13. An analysis device for analyzing a bleeding tendency of a subject, comprising: The analysis device includes a control device, The control device includes: obtaining a time series data set relating to a coagulation reaction of a blood sample taken from the subject; Based on the acquired time series data set, a maximum velocity arrival time until the velocity of the coagulation reaction becomes maximum, or a maximum acceleration arrival time until the acceleration of the coagulation reaction becomes maximum, and a reference time in the coagulation reaction are acquired; Obtaining a value based on the acquired maximum velocity arrival time or the acquired maximum acceleration arrival time and the reference time; outputting the acquired value or information regarding bleeding tendency based on the value; The value indicates the degree of bleeding tendency. The analysis device.

14. An analysis device according to claim 13; a measuring device that irradiates light onto the blood sample in which a coagulation reaction has started and acquires optical information; The control device acquires the time series data set based on the optical information. Analysis system.

15. When the program is executed by a computer, the computer Obtaining a time series data set relating to a coagulation reaction of a blood sample taken from a subject; A step of acquiring a maximum velocity arrival time until the velocity of the coagulation reaction becomes maximum, or a maximum acceleration arrival time until the acceleration of the coagulation reaction becomes maximum, and a reference time in the coagulation reaction based on the acquired time series data set; acquiring a value based on the acquired maximum velocity arrival time or the acquired maximum acceleration arrival time and the reference time; and outputting the acquired value or information related to bleeding tendency based on the value; The value indicates the degree of bleeding tendency. An analysis program that analyzes a subject's tendency to bleed.

Images