Sample analysis method and sample analysis device

Abstract

Provided are a test specimen analysis method and a test specimen analysis device that support rapid evaluation of the severity of an organ disorder caused by an infectious disease, and a test specimen analysis method and a test specimen analysis device that are capable of outputting information highly related to a score indicating the severity of an organ disorder caused by an infectious disease. The blood cell analyzer (1) includes a measurement unit (measurement unit (2)) that irradiates a measurement sample containing a blood specimen and a fluorescent dye for staining leukocytes contained in the blood specimen with light, detects fluorescence of the leukocytes contained in the measurement sample irradiated with the light, and measures a fluorescence signal on the basis of the detected fluorescence. The method comprises the following steps: generating information related to the distribution width of the fluorescence intensity of a neutrophil group contained in a measurement sample, and generating information related to the distribution width of the fluorescence intensity of the neutrophil group based on the information related to the distribution width of the fluorescence intensity of the neutrophil group; an information processing unit (main body 40) that specifies severity-related information of the organ disorder caused by the infectious disease; and an output unit 42 that outputs the severity-related information of the organ disorder caused by the infectious disease.

Description

[Technical field]

[0001] The present invention relates to a subject analysis method and a subject analysis device for analyzing a subject. [Background technology]

[0002] Non-patent document 1 discloses that among the data of various leukocytes obtained from the multi-item automatic blood cell analyzer XN-9000 (manufactured by Sysmex Corporation), the fluorescence intensity (NE-SFL) obtained from neutrophils with an average nucleic acid amount and the scattered light intensity (MO-X) obtained from monocytes with average morphological characteristics are correlated with the sequential organ failure assessment (SOFA) score indicating the degree of organ dysfunction.

[0003] [Prior art literature]

[0004]

Non-patent literature

[0005] [Non-patent document 1] Sabrina Buoro, et al. "Clinical significance of cellpopulation data (CPD) on Sysmex XN-9000in septic patients with our without liver impairment", Annals of Translational Medicine, November 2016, Vol4, No 21, p.1-9

[0006] [Summary of the Invention]

[0007] [Problems to be solved by the invention]

[0008] The SOFA score is a method for evaluating the severity of organ dysfunction caused by infectious diseases advocated by ESICM (European society of intensive care medicine) and is widely used in ICUs. In order to calculate the SOFA score, it is necessary to perform multiple tests including ventilator, coagulation energy, liver, circulatory system, central nervous system, and kidney function, and the calculation of the score requires a long time. Therefore, as a faster inspection method, a method using NE-SFL and MO-X obtained from the multi-item automatic blood cell analyzer XN-9000 is proposed in non-patent document 1.

[0009] However, the Spearman's rank correlation coefficient between NE-SFL and MO-X and SOFA score shown in Non-Patent Document 1 is less than 0.4, and it is not feasible to use NE-SFL and MO-X as a method for evaluating the severity of organ dysfunction caused by infections in clinical settings.

[0010] Therefore, there is a need for a subject analysis method and a subject analysis device that support rapid evaluation of the severity of organ damage caused by an infection, and that can output information highly correlated with a score indicating the severity of organ damage caused by an infection.

[0011]

Methods for solving the problem

[0012] The subject analysis method of the present invention includes: irradiating light onto a measurement sample containing a blood subject and a fluorescent dye that stains leukocytes contained in the above-mentioned blood subject, detecting the fluorescence of the leukocytes contained in the above-mentioned measurement sample irradiated with the above-mentioned light, and a measurement step of measuring a fluorescence signal based on the detected fluorescence, a generation step of generating information related to the distribution width of the fluorescence intensity of a neutrophil group contained in the above-mentioned measurement sample based on the above-mentioned fluorescence signal, a determination step of determining information related to the severity of an organ disorder caused by an infectious disease based on the information related to the distribution width of the fluorescence intensity of the above-mentioned neutrophil group, and an output step of outputting information related to the severity of the organ disorder caused by the above-mentioned infectious disease.

[0013] The subject analysis device (1) of the present invention comprises: a measuring unit (2) for irradiating light onto a measurement sample containing a blood subject and a fluorescent dye for staining leukocytes contained in the blood subject, detecting the fluorescence of the leukocytes contained in the measurement sample irradiated with light, and measuring a fluorescence signal based on the detected fluorescence; an information processing unit (40) for generating information related to the distribution width of the fluorescence intensity of a neutrophil group contained in the measurement sample based on the measured fluorescence signal; determining information related to the severity of an organ disorder caused by an infectious disease based on the information related to the distribution width of the fluorescence intensity of the neutrophil group; and an output unit (42) for outputting information related to the severity of the organ disorder caused by the infectious disease.

[0014] [Effects of the invention]

[0015] The present invention can provide a subject analysis method and a subject analysis device that support rapid evaluation of the severity of organ damage caused by an infection, which are a subject analysis method and a subject analysis device that can output information highly correlated with a score indicating the severity of organ damage caused by an infection.

[0016] [Brief description of the attached drawings]

[0017]

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[0018]

Figure 2

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Figure 4

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Figure 5A

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Figure 5B

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[0033]

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[0034]

Fig.16

[0035] The subject analysis device according to the embodiment will be described with reference to the drawings. The subject analysis device according to the embodiment is a blood cell analysis device that analyzes blood cells of a blood subject.

[0036] [1. First embodiment]

[0037] [1-1. Schematic structure]

[0038] Figure 1 1 is an oblique view showing the appearance of the blood cell analyzer involved in the first embodiment. The blood cell analyzer 1 is a device for analyzing the blood cells of a blood subject. Specifically, the blood cell analyzer 1 is a multi-item blood cell analyzer that classifies the types of white blood cells contained in the blood subject, counts the number of various white blood cells, counts red blood cells and platelets, and measures the hemoglobin concentration. Furthermore, in the following description, the description of the counting of red blood cells and platelets, and the measurement of hemoglobin concentration is omitted. In the counting of red blood cells and platelets, and the measurement of hemoglobin concentration, for example, the measurement unit described in the gazette of U.S. Patent No. 8,968,653 can be used, and the gazette is integrated into this specification by reference.

[0039] The blood cell analyzer 1 includes a measuring unit 2 , a transport unit 3 , and an information processing unit 4 .

[0040] The transport unit 3 is disposed in front of the measuring unit 2 and supplies the blood sample to the measuring unit 2. Specifically, the transport unit 3 moves to a position P1 in front of the measuring unit 2 (see Figure 2 ) A sample rack L is used to transport and support a plurality of sample containers (blood collection tubes) T containing blood samples. The blood sample is peripheral blood collected from a subject. The subject is a person from whom peripheral blood is collected, for example, a patient. The patient is, for example, a person who is likely to develop an organ disorder caused by an infectious disease.

[0041] The measuring unit 2 mixes a reagent containing a fluorescent dye for staining leukocytes with a blood sample to prepare a measuring sample used in the measurement. The measuring unit 2 irradiates light to the prepared measuring sample to measure the scattered light signal and the fluorescent signal of the leukocytes contained in the measuring sample. The measuring unit 2 can be connected to the information processing unit 4 for communication and sends the measurement result to the information processing unit 4.

[0042] The information processing unit 4 can communicate with the measuring unit 2, the transport unit 3, and the host computer 5 (see Figure 2 The information processing unit 4 controls the operation of the measuring unit 2 and the transport unit 3, performs analysis based on the measurement result of the measuring unit 2, and outputs the analysis result through the output unit 42 and sends it to the host computer 5. The information processing unit 4 includes a main body 40 (see Figure 4 ), an input unit 41 and an output unit 42. The input unit 41 is, for example, a keyboard and a mouse. The output unit 42 is, for example, a display. The display can be, for example, a liquid crystal display, an organic EL display, or a small LED display. The input unit 41 and the output unit 42 can also be touch panels.

[0043] [1-2. Detailed structure]

[0044] (Measurement unit)

[0045] Figure 2 The measuring unit 2 includes a hand 21 , a sample container setting unit 22 , a barcode unit 23 , a sample suction unit 24 , a sample preparation unit 25 , and a detection unit 26 . Each of the units 21 to 26 is controlled by the information processing unit 4 .

[0046] The hand 21 holds the sample container T located at the position P1, pulls out the sample container T from the sample rack L and shakes it. The sample container setting part 22 receives the sample container T shaken by the hand 21 above the position P1. The barcode unit 23 reads the sample number from the barcode label attached to the sample container T. The sample number is information associated with the analysis result of the blood sample. The sample suction part 24 is equipped with a suction tube 24a, and sucks the blood sample from the sample container T. The sample preparation part 25 is equipped with a mixing chamber MC and a heater H, and prepares the measurement sample used in the measurement by mixing the reagent with the blood sample (hereinafter, sometimes simply referred to as "sample") in the sample container T. The detection part 26 is equipped with an optical detector D, and detects the fluorescence and scattered light of white blood cells from the measurement sample.

[0047] The operation of the measurement unit 2 will be described, and the sample preparation section 25 and the detection section 26 will be described in detail.

[0048] The sample container T located at the position P1 by the transport unit 3 is grasped by the hand 21 and pulled upward from the sample rack L. Then, the sample in the sample container T is stirred by shaking the hand 21. The sample container T after stirring is set by the hand 21 to the sample container setting part 22 located above the position P1. Then, the sample container T is transported to the position P2 by the sample container setting part 22.

[0049] When the sample container T is located at the position P2, the sample number is read from the barcode label attached to the sample container T by the barcode unit 23 provided near the position P2. Then, the sample container T is transported to the position P3 by the sample container setting unit 22. When the sample container T is located at the position P3, a specified amount of the sample is sucked from the sample container T via the suction tube 24a by the sample suction unit 24. When the suction of the sample is completed, the sample container T is transported forward by the sample container setting unit 22 and returned to the original support position of the sample rack L by the hand 21. After the sample sucked through the suction tube 24a is moved to the position of the mixing chamber MC, only a specified amount is discharged into the mixing chamber MC by the sample suction unit 24.

[0050] The sample preparation section 25 is connected to a reagent container 251 containing a first reagent, a reagent container 252 containing a second reagent, and a reagent container 253 containing a sheath liquid (diluent) via a tube. In addition, the sample preparation section 25 is connected to a compressor (not shown), and the pressure generated by the compressor enables the respective reagents to be separated from the reagent containers 251 to 253. The sample preparation section 25 mixes the blood sample and the reagent in the mixing chamber MC, and heats the mixed solution by the heater H for a specified time to prepare a measurement sample. The measurement sample prepared by the sample preparation section 25 is supplied to the optical detector D of the detection section 26.

[0051] The reagent includes a first reagent for staining nucleic acid of leukocytes and a second reagent for hemolyzing erythrocytes and damaging the cell membrane of leukocytes to such an extent that the fluorescent dye can penetrate.

[0052] The first reagent contains a fluorescent dye capable of staining nucleic acid and is used to fluorescently stain nucleic acid of nucleated cells such as leukocytes in a blood sample treated with the second reagent described below. By treating the blood sample with the first reagent, blood cells having nucleic acid such as leukocytes are stained.

[0053] The fluorescent dye is not particularly limited as long as it can dye nucleic acid, and can be appropriately selected corresponding to the wavelength of light irradiated from the light source (the semiconductor laser D31 described later). As such fluorescent dyes, for example, propidium iodide, ethidium bromide, ethidium-acridine heterodimer, ethidium diazide, ethidium homodimer-1, ethidium homodimer-2, ethidium monoazide, trimethylenebis[[3-[[4-[[(3-methylbenzothiazol-3-ium)-2-yl]methylene]-1,4-dihydroquinoline]-1-yl]propyl]dimethylammonium] tetraiodide (TOTO-1), 4-[(3-methylbenzothiazol-2(3H)-ylidene)methyl]-1-[3-(trimethylammonium)propyl]quinolinium diiodide (T O-PRO-1), N,N,N',N'-tetramethyl-N,N'-bis[3-[4-[3-[(3-methylbenzothiazol-3-ium)-2-yl]-2-propenyl]-1,4-dihydroquinolin-1-yl]propyl]-1,3-propanediammonium tetraiodide (TOTO-3), or 2-[3-[[1-[3-(trimethylammonium)propyl]-1,4-dihydroquinolin]-4-ylidene]-1-propenyl]-3-methylbenzothiazol-3-ium diiodide (TO-PRO-3) and a fluorescent dye represented by the following general formula (I). Among them, the fluorescent dye represented by the general formula (I) is also preferred.

[0054]

Chemistry 1

[0055]

[0056] In the above formula (I), R1 and R4 are the same or different from each other and are hydrogen atoms, alkyl groups, alkyl chains having hydroxyl groups, alkyl chains having ether groups, alkyl chains having ester groups, or benzyl groups optionally having substituents. In addition, R2 and R3 are the same or different from each other and are hydrogen atoms, hydroxyl groups, halogen groups, alkyl groups, alkenyl groups, alkynyl groups, alkoxy groups, alkylsulfonyl groups, or phenyl groups. In addition, Z is a sulfur atom, an oxygen atom, or a carbon atom having a methyl group, n is 0, 1, 2, or 3, and X is - It is an anion.

[0057] In the present embodiment, the alkyl group may be either a straight chain or a branched chain. In addition, in the above formula (I), when either R1 or R4 is an alkyl group having 6 to 18 carbon atoms, the other is preferably a hydrogen atom or an alkyl group having less than 6 carbon atoms. Among the alkyl groups having 6 to 18 carbon atoms, an alkyl group having 6, 8 or 10 carbon atoms is also preferred.

[0058] In the above formula (I), examples of the substituent of the benzyl group of R1 and R4 include an alkyl group having 1 to 20 carbon atoms, an alkenyl group having 2 to 20 carbon atoms, or an alkynyl group having 2 to 20 carbon atoms. Among them, a methyl group or an ethyl group is particularly preferred.

[0059] In the above formula (I), examples of the alkenyl groups for R2 and R3 include alkenyl groups having 2 to 20 carbon atoms. Examples of the alkoxy groups for R2 and R3 include alkoxy groups having 1 to 20 carbon atoms. Among them, methoxy and ethoxy groups are particularly preferred.

[0060] In the above formula (I), the anion X - , such as F - , Cl - Br - and I - Same halogen ion, CF3SO3 - 、BF4 - wait.

[0061] In addition, the fluorescent dye in the first reagent may be one kind or two or more kinds.

[0062] In addition, the concentration of the fluorescent dye in the first reagent can be appropriately set according to the type of the fluorescent dye, and is generally 0.01 to 100 pg / μL, preferably 0.1 to 10 pg / μL. For example, when the fluorescent dye represented by the above formula (I) is used as the fluorescent dye of the first reagent, the concentration of the fluorescent dye in the first reagent is preferably 0.2 to 0.6 pg / μL, and more preferably 0.3 to 0.5 pg / μL.

[0063] In addition, the first reagent can be obtained by dissolving the above-mentioned fluorescent dye in a suitable solvent in a mode of becoming the above-mentioned concentration. As a solvent, as long as the above-mentioned fluorescent dye can be dissolved, it is not particularly limited, and for example, water, an organic solvent and a mixture thereof can be mentioned. As an organic solvent, for example, alcohol, ethylene glycol, dimethyl sulfoxide (DMSO) etc. can be mentioned. Furthermore, since the fluorescent dye has a poor storage stability in an aqueous solution, it is preferably dissolved in an organic solvent.

[0064] In addition, a commercially available staining reagent for leukocyte classification may be used as the first reagent. Examples of such a staining reagent include FLUOROCELL WDF manufactured by Sysmex Corporation.

[0065] The second reagent contains a surfactant for hemolyzing red blood cells and damaging the cell membrane of white blood cells to the extent that the fluorescent dye in the first reagent can pass through, for example, at least one of a cationic surfactant, a nonionic surfactant, and an anionic surfactant, or a combination thereof. Furthermore, the second reagent preferably contains an aromatic organic acid at a concentration of 20 mM to 50 mM. When the concentration of the aromatic organic acid in the second reagent is 20 mM to 30 mM, the pH of the second reagent is 5.5 to 6.4, and when the concentration of the aromatic organic acid is 30 mM to 50 mM, the pH of the second reagent is preferably 5.5 to 7.0.

[0066] In this embodiment, when the concentration of the aromatic organic acid in the second reagent is 20 mM or more and less than 30 mM, the pH of the second reagent is preferably 5.5 or more and 6.4 or less, and more preferably 5.5 or more and 6.2 or less. In addition, when the concentration of the aromatic organic acid in the second reagent is 30 mM or more and 50 mM or less, and preferably 40 mM or more and 50 mM or less, the pH of the second reagent is preferably 5.5 or more and 7.0 or less. More preferably, when the concentration of the aromatic organic acid in the second reagent is 40 mM or more and 50 mM or less, the pH of the second reagent is preferably 5.5 or more and 6.2 or less.

[0067] In addition, in the present embodiment, the aromatic organic acid refers to an acid and a salt thereof having at least one aromatic ring in the molecule. As the aromatic organic acid, for example, aromatic carboxylic acids, aromatic sulfonic acids, etc. can be cited. In the present embodiment, phthalic acid, benzoic acid, salicylic acid, hippuric acid, p-aminobenzenesulfonic acid, benzenesulfonic acid and their salts are suitably used. Furthermore, the aromatic organic acid in the second reagent can be one kind or two or more kinds. When the second reagent contains two or more aromatic organic acids, the total of their concentrations can also be 20 mM or more and 50 mM or less.

[0068] As the cationic surfactant, for example, a quaternary ammonium salt type surfactant or a pyridinium salt type surfactant can be used.

[0069] Examples of the quaternary ammonium salt surfactant include surfactants represented by the following formula (II) and having a total carbon number of 9 to 30.

[0070]

Chemistry 2

[0071]

[0072] In the above formula (II), R1 is an alkyl or alkenyl group having 6 to 18 carbon atoms. R2 and R3 are the same or different from each other and are alkyl or alkenyl groups having 1 to 4 carbon atoms. R4 is an alkyl or alkenyl group having 1 to 4 carbon atoms, or a benzyl group. X - It is a halogen atom.

[0073] In the above formula (II), R1 is preferably an alkyl or alkenyl group having 6, 8, 10, 12 or 14 carbon atoms, and a straight-chain alkyl group is particularly preferred. More specifically, octyl, decyl and dodecyl groups can be cited. R2 and R3 are preferably methyl, ethyl and propyl groups. R4 is preferably methyl, ethyl and propyl groups.

[0074] Examples of the pyridinium salt-based surfactant include surfactants represented by the following formula (III).

[0075]

Chemistry 3

[0076]

[0077] In the above formula (III), R1 is an alkyl or alkenyl group having 6 to 18 carbon atoms, and X - It is a halogen atom.

[0078] In the above formula (III), R1 is preferably an alkyl or alkenyl group having 6, 8, 10, 12 or 14 carbon atoms, and a linear alkyl group is particularly preferred. More specific examples of R1 include octyl, decyl and dodecyl.

[0079] The concentration of the cationic surfactant in the second agent can be appropriately adjusted depending on the type of the surfactant, and is usually 10 to 10,000 ppm, preferably 100 to 1,000 ppm.

[0080] As the nonionic surfactant, a polyoxyethylene nonionic surfactant represented by the following formula (VI) is preferred.

[0081] R1-R2-(CH2CH2O) n -H(VI)

[0082] In the above formula (VI), R1 is an alkyl, alkenyl or alkynyl group having 8 to 25 carbon atoms, and R2 is an oxygen atom, -COO- or

[0083]

Chemistry 4

[0084]

[0085] n is an integer from 10 to 50.

[0086] Specific examples of the nonionic surfactant include polyoxyethylene alkyl ether, polyoxyethylene sterol, polyoxyethylene castor oil, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene alkylamine, polyoxyethylene polyoxypropylene alkyl ether, and the like.

[0087] The concentration of the nonionic surfactant in the second agent is usually 10 to 100,000 ppm, preferably 100 to 10,000 ppm, and more preferably 1,000 to 5,000 ppm.

[0088] The second reagent may also contain a buffer in order to keep the pH constant. Such a buffer may include, for example, citrate, HEPES, and phosphate. Furthermore, the above-mentioned aromatic organic acid may exhibit a buffering effect. When such an aromatic organic acid is used, a buffer may be added to the second reagent at will.

[0089] The osmotic pressure of the second reagent is not particularly limited, but is preferably 20 to 150 mOsm / kg from the viewpoint of effectively hemolyzing erythrocytes.

[0090] In addition, the second reagent can be obtained by dissolving the above-mentioned surfactant and aromatic organic acid or its salt and the above-mentioned buffer agent in a suitable solvent in a manner to become the above-mentioned aromatic organic acid concentration according to the expectation, and adjusting the pH using NaOH, HCl, etc. As the solvent, as long as the above-mentioned components can be dissolved, it is not particularly limited, and for example, water, organic solvents and mixtures thereof can be cited. As the organic solvent, for example, alcohol, methanol, ethylene glycol, DMSO, etc. can be cited.

[0091] In addition, as the second reagent, a commercially available hemolytic reagent for leukocyte classification may be used. Examples of such a hemolytic reagent include LYSERCELL WDF and LYSERCELL WDF II manufactured by Sysmex Corporation.

[0092] The second reagent constructed as described above lyses red blood cells and damages the cell membrane of white blood cells to the extent that the fluorescent dye in the first reagent can pass through. In addition, the nucleic acid of the white blood cells whose cell membranes are damaged by the second reagent is stained by the first reagent constructed as described above. Thus, it becomes possible to detect the four subtypes of white blood cells by the optical detector D.

[0093] The detection unit 26 is connected to a reagent container 253 containing a sheath liquid (diluent) via a tube. In addition, the detection unit 26 is connected to a compressor (not shown), and the pressure generated by the compressor enables the sheath liquid (diluent) to be separated from the reagent container 253. The separated sheath liquid is used in the optical detector D.

[0094] Figure 3: is a diagram showing the structure of an optical detector. The optical detector D is a flow cytometer using a semiconductor laser, and is equipped with a flow cell D1, a sheath flow system D2, a beam spot forming system D3, a forward scattered light receiving system D4, a side scattered light receiving system D5, a fluorescence light receiving system D6, and a signal processing circuit D7.

[0095] The flow cell D1 is a tubular component made of a light-transmitting material. The sheath flow system D2 is configured to send the measurement sample wrapped in sheath liquid into the flow cell D1 to generate a liquid flow in the flow cell D1. The beam spot forming system D3 is configured to irradiate the flow cell D1 with light emitted from a semiconductor laser D31 as a light source through a collimator lens D32 and a condenser lens D33. Thus, the laser is irradiated to the blood cells contained in the liquid flow passing through the flow cell D1. In addition, the beam spot forming system D3 is further provided with a beam stopper D34.

[0096] The forward scattered light receiving system D4 is configured such that the light (forward scattered light) scattered forward by the blood cells irradiated with the laser is condensed by the front condensing lens D41, and the light passing through the pinhole D42 is received by the photodiode D43. The photodiode D43 outputs an electric signal having a size corresponding to the intensity of the received light. The electric signal output from the photodiode D43 is sampled multiple times by the signal processing circuit D7 between each blood cell passing through the laser irradiation area in the flow cell D1, converted into a digital signal, and sent to the information processing unit 4. This digital signal (waveform signal) is the measurement result of the received forward scattered light, and shows the forward scattered light signal.

[0097] The side scattered light receiving system D5 is configured to collect light (side scattered light) scattered to the side by the blood cells irradiated with laser light by the side condensing lens D51, reflect it by the dichroic mirror D52, and receive it by the photodiode D53. The photodiode D53 outputs an electric signal having a size corresponding to the intensity of the received light. The electric signal output from the photodiode D53 is sampled multiple times by the signal processing circuit D7 between each blood cell passing through the laser irradiation area in the flow cell D1, converted into a digital signal, and sent to the information processing unit 4. This digital signal (waveform signal) is the measurement result of the received side scattered light, and shows the side scattered light signal.

[0098] Light scattering is a phenomenon that occurs when particles such as blood cells exist as obstacles in the direction of light travel, and the light changes its direction of travel due to the particles. By detecting this scattered light, it is possible to obtain information related to the size and material of the particles. In particular, information related to the size of the particles (blood cells) can be obtained from the forward scattered light. Specifically, the higher the intensity of the forward scattered light, the larger the size of the cell (such as white blood cells). In addition, information about the inside of the particle can be obtained from the side scattered light. When the blood cell particles are irradiated with laser light, the intensity of the side scattered light depends on the complexity of the inside of the cell (the shape, size, density and amount of particles of the nucleus). For example, the higher the intensity of the side scattered light, the higher the density of the cell (blood cell) and the greater the amount of particles contained in the cell (blood cell). In addition, the higher the intensity of the side scattered light, the more complex the shape of the nucleus and the more lobes the nucleus has.

[0099] The fluorescence light receiving system D6 is constructed in such a way that the fluorescence emitted by irradiating the blood cells stained with nucleic acid with laser light is received by the photodiode D62 as an avalanche photodiode through the dichroic mirror D52 and the spectroscopic filter D61. The photodiode D62 outputs an electrical signal having a size corresponding to the intensity of the received light. The electrical signal output from the photodiode D62 is sampled multiple times by the signal processing circuit D7 between each blood cell passing through the laser irradiation area in the flow cell D1, converted into a digital signal, and sent to the information processing unit 4. This digital signal (waveform signal) is the measurement result of the received fluorescence and shows the fluorescence signal. Furthermore, the photodiode D62 does not necessarily have to be an avalanche photodiode, and for example, it can also be a photomultiplier tube.

[0100] When light is irradiated onto blood cells with nucleic acid stained with a fluorescent substance, the fluorescent substance fluoresces. Since the intensity of the fluorescence becomes stronger as the amount of stained nucleic acid increases, blood cells can be classified based on the difference in the amount of nucleic acid contained in the blood cells by measuring the fluorescent signal.

[0101] return Figure 2 The measurement results of the forward scattered light signal, the side scattered light signal and the fluorescence signal measured by the optical detector D are sent to the information processing unit 4.

[0102] As described above, the measuring unit 2 functions as a modulation unit for modulating a blood subject and a measuring sample containing a fluorescent dye for staining white blood cells contained in the blood subject, and a measuring unit for irradiating light to the measuring sample, detecting the fluorescence of the white blood cells contained in the measuring sample irradiated with light, and measuring a fluorescence signal based on the detected fluorescence.

[0103] (Information Processing Unit)

[0104] Figure 42 is a diagram showing the configuration of the information processing unit. The information processing unit 4 is configured including a computer, and includes a main body 40, an input unit 41, and an output unit 42. The description of the input unit 41 and the output unit 42 is omitted as described above.

[0105] The main body 40 includes a CPU 401 , a ROM 402 , a RAM 403 , a storage device 404 , a reading device 405 , an image output interface 406 , an input / output interface 407 , and a communication interface 408 .

[0106] The CPU 401 executes computer programs stored in the ROM 402 and computer programs loaded in the RAM 403. The RAM 403 is used to read computer programs recorded in the ROM 402 and the storage device 404. In addition, the RAM 403 is also used as a work area of ​​the CPU 401 when executing these computer programs.

[0107] The storage device 404 is a solid state drive (SSD), a hard disk drive (HDD), or a combination thereof. The storage device 404 stores an operating system (OS), a computer program to be executed by the CPU 401, and data used in the execution of the computer program. In addition, the storage device 404 stores data used to execute the computer program. Fig.11 The reading device 405 may be composed of a CD drive or a DVD drive, etc., and reads the computer program and data recorded on the recording medium 405a. Furthermore, when the above-mentioned program 404a is recorded on the recording medium 405a, the program 404a read from the recording medium 405a by the reading device 405 is stored in the storage device 404.

[0108] The image output interface 406 outputs an image signal corresponding to the data to the output unit 42, and the output unit 42 displays an image based on the data output from the image output interface 406. The user inputs instructions via the input unit 41, and the input / output interface 407 receives the signal input via the input unit 41. The communication interface 408 is connected to the measurement unit 2, the transport unit 3, and the host computer 5, and the CPU 401 transmits and receives control signals and data with these devices via the communication interface 408.

[0109] The main body 40 is composed of components 401 to 408. When the CPU 401 executes the user's instructions and / or computer programs through the input unit 41, it functions as a control unit for controlling the measuring unit 2 and the conveying unit 3. At the same time, when the CPU 401 executes the computer program, it functions as an information processing unit.

[0110] In other words, the information processing unit 4 controls the operation of the measuring unit 2 and the transport unit 3 by means of the main body 40, and performs analysis based on the measurement results (e.g., forward scattered light signals, side scattered light signals, and fluorescent signals) received from the measuring unit 2. In this analysis, the leukocytes present in the blood sample are classified into four subtypes, namely, (1) lymphocytes (LYMPH), (2) monocytes (MONO), (3) a blood cell group consisting of neutrophils (NEUT) and basophils (BASO), and (4) eosinophils (EO). The classification procedure will be described below.

[0111] The information processing unit 4 extracts a digital signal representing the peak value of each blood cell from each of the forward scattered light signals (digital signals sampled multiple times while each blood cell passes through the laser irradiation area in the flow cell D1) received from the measuring unit 2, and obtains the extracted digital signal as the forward scattered light intensity of each blood cell. Similarly, the information processing unit 4 extracts a digital signal representing the peak value of each blood cell from each of the side scattered light signals of each blood cell, and obtains the extracted digital signal as the side scattered light intensity of each blood cell, and extracts a digital signal representing the peak value of each blood cell from each of the fluorescence signals of each blood cell, and obtains the extracted digital signal as the fluorescence intensity of each blood cell. Thereafter, the information processing unit 4 generates a scattergram. The scattergram is a distribution diagram of each blood cell plotted based on the forward scattered light intensity, the side scattered light intensity, and the fluorescence intensity. The information processing unit 4 can generate a three-dimensional scattergram with the forward scattered light intensity, the side scattered light intensity, and the fluorescence intensity as axes, and a two-dimensional scattergram with the side scattered light intensity and the fluorescence intensity as axes. The scattergram of this embodiment is a two-dimensional scattergram.

[0112] Figure 5A is a diagram showing an example of a scatter plot. Figure 5A As shown, the scattergram of the present embodiment is generated by plotting positions corresponding to the side scattered light intensity and fluorescence intensity of each blood cell measured by the measuring unit 2 on a two-dimensional graph with the horizontal axis being the side scattered light intensity (SSC) and the vertical axis being the fluorescence intensity (SFL).

[0113] Next, the information processing unit 4 performs fractionation processing on the scattergram by the main body 40. The fractionation processing is a process of classifying the white blood cells in the scattergram into any clusters and distinguishing them on the scattergram. The procedure and method of the fractionation processing will be described.

[0114] Figure 5B: is a diagram showing an example of a scatter plot that has been subjected to fractionation processing. First, the information processing unit 4 removes points corresponding to fragments composed of hemolyzed red blood cells, etc. from the points plotted on the scatter plot. Specifically, the points included in the predetermined designated area DB are removed. Next, the information processing unit 4 classifies the points on the scatter plot of blood cells excluding the area DB into any one of the four clusters (groups) corresponding to the four subclasses of white blood cells. Figure 5B As shown, the four clusters are (1) cluster A11 of lymphocytes (LYMPH), (2) cluster A12 of monocytes (MONO), (3) cluster A13 of a blood cell group consisting of neutrophils (NEUT) and basophils (BASO), and (4) cluster A14 of eosinophils (EO).

[0115] Specifically, the information processing unit 4 calculates the degree of belonging to the cluster of each blood cell from the distance between the point corresponding to each blood cell plotted on the scattergram and the center of gravity position of each cluster set in advance, and assigns each blood cell to each cluster according to the degree of belonging. Such a method of classifying each blood cell is described in detail in U.S. Patent No. 5555196, which is incorporated into this specification.

[0116] When each blood cell is classified into any of the four clusters corresponding to the four subclasses of white blood cells, each point plotted on the scatter plot becomes as follows: Figure 5B The clusters A11 to A14 shown are similarly distinguished.

[0117] The information processing unit 4 counts the number of blood cells in each cluster. Specifically, the information processing unit 4 counts the number of points contained in each cluster of clusters A11 to A14 (the number of blood cells belonging to each cluster) by the main body 40. Thus, the number of lymphocytes, monocytes, blood cell groups composed of neutrophils and basophils, and eosinophils is obtained. In addition, the information processing unit 4 calculates the number of white blood cells counted by the main body 40 (that is, the total number of points contained in each cluster of clusters A11 to A14), the ratio of lymphocytes, monocytes, blood cell groups composed of neutrophils and basophils, and eosinophils to white blood cells.

[0118] The information processing unit 4 generates information related to the distribution width of the fluorescence intensity of the neutrophil group based on the fluorescence intensity of the blood cells belonging to the cluster A13 by the main body 40. The neutrophil group is a blood cell group composed of neutrophils and basophils, and is a blood cell group contained in the cluster A13. In general, since peripheral blood contains more neutrophils than basophils, the blood cell group of the cluster A13 can be captured as a group of neutrophils. Furthermore, blood cells corresponding to basophils can be excluded from the blood cell group of the cluster A13, and the remaining blood cell group can be used as the neutrophil group.

[0119] The information related to the distribution width of the fluorescence intensity of the neutrophil group is an index indicating the distribution width of the fluorescence intensity of the neutrophil group. As an index indicating the distribution width of the fluorescence intensity of the neutrophil group, (1) a distribution width of the fluorescence intensity of the neutrophil group passing through the center of gravity G of the neutrophil group (refer to Figure 5B ) is the distance DW from the upper limit of the Y axis to the lower limit of the Y axis of the neutrophil group on the X axis (refer to Figure 5B ), (2) the distance DW1 from the center of gravity G to the upper limit of the Y axis of the neutrophil group on the X axis passing through the center of gravity G (refer to Figure 5B ), (3) the distance DW2 from the center of gravity G to the lower limit of the Y axis of the neutrophil group on the X axis passing through the center of gravity G (refer to Figure 5B ), (4) the average of the distance DW1 from the center of gravity G to the upper limit of the Y axis of the neutrophil group on the X axis passing through the center of gravity G and the distance DW2 from the center of gravity G to the lower limit of the Y axis of the neutrophil group on the X axis passing through the center of gravity G. The information processing unit 4 calculates the center of gravity G by the main body 40. In addition, as an index representing the distribution width of the fluorescence intensity of the neutrophil group, it is also possible to use (5) the distance in the Y axis direction between the blood cell with the maximum fluorescence intensity of the neutrophil group and the blood cell with the minimum fluorescence intensity, regardless of the center of gravity G. As information related to the distribution width of the fluorescence intensity of the neutrophil group, for example, NE-WY output by the same method as the multi-item automatic blood cell analyzer XN series (XN-1000, XN-2000, etc.) manufactured by Sysmex Corporation can also be used.

[0120] The inventors have found that the distribution width of the fluorescence intensity of the neutrophil group is highly correlated with the SOFA score (Sequential Organ Failure Assessment Score) used in the evaluation of the severity of organ failure caused by an infection. In addition, by studying the SOFA score of patients in the early stages of hospitalization, the inventors have found that the distribution width of the fluorescence intensity of the neutrophil group tends to increase in patients with severe organ failure caused by an infection. The experiments and analyses that have yielded these insights are described below.

[0121] The present inventors analyzed blood samples of hospitalized patients suspected of sepsis on each day of the initial stage of hospitalization and blood samples of healthy subjects by the method using the blood cell analyzer 1 described above. In this analysis, blood samples of 44 hospitalized patients suspected of sepsis and 39 healthy subjects were used. The details of hospitalized patients suspected of sepsis included 23 sepsis patients diagnosed with sepsis after hospitalization and 21 non-septic patients suspected of sepsis but not sepsis.

[0122] The selection criteria for hospitalized patients suspected of having sepsis are those with fever (38°C or above) and qSOFA (quickSOFA) score of 2 or above. However, patients who are taking steroids are clearly stated. qSOFA is a simpler sepsis screening method than SOFA, consisting of three items: respiratory rate, mental state, and systolic blood pressure. The exclusion criteria for hospitalized patients suspected of having sepsis are patients in the final stage of chronic diseases, patients with neutropenia, dialysis patients, Actemra (registered trademark) (humanized anti-human IL6 receptor monoclonal antibody), and patients taking TNFα inhibitors.

[0123] The selection criteria for healthy subjects were: no fever, no suspected infection, no chronic inflammatory disease, no abnormal coagulation ability, and no oral anticoagulant. The exclusion criteria for healthy subjects were: no.

[0124] Specifically, the present inventors collected blood samples from hospitalized patients suspected of having sepsis on the first day of hospitalization (day 0), day 1, and day 3, which are the initial stage of hospitalization, and collected blood samples only once from healthy subjects. Furthermore, an examination for obtaining a SOFA score for each hospitalized patient was performed to obtain a SOFA score for each day (day 0, day 1, and day 3). In addition, the present inventors used the collected blood samples to modulate the measurement samples of the hospitalized patients on each day and the measurement samples of the healthy subjects by the method using the measurement unit 2, measured the side scattered light signal and the fluorescence signal, and calculated the white blood cell related parameters including the distribution width related information (NE-WY) of the fluorescence intensity of the neutrophil group by the method of the information processing unit 4. In these modulations, the measurement of the side scattered light signal and the fluorescence signal, and the calculation of the white blood cell related parameters including NE-WY, the multi-item automatic blood cell analyzer XN-2000 manufactured by Sysmex Co., Ltd. was used.

[0125] Figure 6 This is a graph showing information related to the distribution width of fluorescence intensity of neutrophil population (NE-WY) for hospitalized patients with sepsis (septic patients), hospitalized patients without sepsis (non-septic patients), and healthy subjects with respect to the number of days from hospitalization. Figure 6 The coordinate plot is a box plot. Figure 6 The horizontal lines shown on the upper side of the graph of indicate two groups whose P values ​​are less than 0.05 when the difference between the data groups is tested by the Steel-Dwass method. For example, the horizontal line in the first row from the top indicates that the P values ​​of the NE-WY data group of healthy subjects and the NE-WY data group of sepsis patients on the third day of hospitalization are less than 0.05 when tested by this method. The horizontal line in the second row from the top indicates that the P values ​​of the NE-WY data group of healthy subjects and the NE-WY data group of sepsis patients on the first day of hospitalization are less than 0.05, and the horizontal line in the third row from the top indicates that the P values ​​of the NE-WY data group of healthy subjects and the NE-WY data group of sepsis patients on the 0th day of hospitalization are less than 0.05. The horizontal line in the sixth row from the top indicates that the P value of the NE-WY data group of non-septic patients on hospital day 0 and the NE-WY data group of septic patients on hospital day 0 is less than 0.05, and the horizontal line in the seventh row from the top indicates that the P value of the NE-WY data group of non-septic patients on hospital day 1 and the NE-WY data group of septic patients on hospital day 1 is less than 0.05. Figure 6As shown, it is known that the NE-WY of the sepsis patient group on the 0th day of hospitalization is significantly greater than the NE-WY of the healthy group and the non-septic patient group on the 0th day of hospitalization. Similarly, it is known that the NE-WY of the sepsis patient group on the 1st day of hospitalization is significantly greater than the NE-WY of the healthy group and the non-septic patient group on the 1st day of hospitalization. In this way, it is known that the NE-WY of the sepsis patient group at the initial stage of hospitalization, which is the 0th day and the 1st day of hospitalization, is significantly greater than the NE-WY of the healthy group and the non-septic patient group at the initial stage of hospitalization. Since the fluorescence intensity is that the higher the fluorescence intensity, the more nucleic acid is expressed, and the large distribution width of the fluorescence intensity of the neutrophil group means that there are neutrophils with a large amount of nucleic acid and neutrophils with a small amount of nucleic acid, which is presumed to indicate that neutrophils have diversity. When sepsis occurs, the reason for the diversity of neutrophils is not limited to this, and it is considered as follows. That is, in sepsis, immature neutrophils are mobilized to peripheral blood. Peripheral blood contains mobilized immature neutrophils and neutrophils that have undergone cell death such as apoptosis or necrosis. Since the amount of nucleic acid contained in immature neutrophils is large and the amount of nucleic acid contained in neutrophils that have undergone cell death is small, it is believed that the distribution width of the fluorescence intensity of the neutrophil group becomes larger in sepsis.

[0126] Figure 7 This is a graph showing the correlation between the distribution width information (NE-WY) of the fluorescence intensity of the neutrophil population and the SOFA score of hospitalized patients (septic patients and non-septic patients) on day 0 of hospitalization. Figure 7 In the figure, white dots are data of sepsis patients, and black dots are data of non-septic patients. The P value of the F test for the significance of regression was less than 0.05. The Spearman's rank correlation coefficient r, which indicates the correlation between NE-WY and SOFA scores, was 0.70, and a high correlation was confirmed between the two. Therefore, it was found that the corresponding SOFA score can be estimated from the information related to the distribution width of the fluorescence intensity of the neutrophil group. In addition, the present inventors Figure 7 The regression equation was calculated using the data of the sepsis patients and non-septic patients shown in . The regression equation is y=0.013x-7.1. However, the x-axis is set to NE-WY and the y-axis is set to SOFA score.

[0127] Figure 8 is a graph showing the correlation between NE-WY and SOFA scores for each day. Figure 8 In the figure, the leftmost coordinate graph is Figure 7 The same coordinate diagram is shown. Figure 8The central coordinate graph and the rightmost coordinate graph in the figure are coordinate graphs showing the correlation between NE-WY and SOFA scores of hospitalized patients (septic patients and non-septic patients) on the 1st day and the 3rd day of hospitalization, respectively. The white dots in each coordinate graph are data of septic patients, and the black dots are data of non-septic patients. The Spearman's rank-order correlation coefficient r, which shows the correlation between NE-WY and SOFA scores, is 0.70 on the first day of hospitalization (day 0), 0.54 on the first day of hospitalization, and 0.27 on the third day of hospitalization. The highest correlation was confirmed on the first day of hospitalization (day 0), and then a high correlation was confirmed on the first day of hospitalization. In addition, it was confirmed that the correlation decreased every day from the start of hospitalization. The reason for the decrease in daily correlation is believed to be that since the treatment was carried out in a manner of suppressing the infection by administering medication from the first day of hospitalization, the infection converged corresponding to the treatment. In addition, the inventors have found that the correlation between NE-WY and SOFA scores ... Figure 8 The regression equations were calculated for the data of sepsis patients and non-septic patients on each day shown in . The regression equations for the first day of hospitalization (day 0), the first day of hospitalization, and the third day of hospitalization were y=0.013x-7.1, y=0.015x-8.3, and y=0.023x-12.1, respectively. However, the x-axis was set to NE-WY, and the y-axis was set to the SOFA score of each day.

[0128] Return to the description of the information processing unit 4. The information processing unit 4, by the main body 40, determines information related to the severity of the organ disorder caused by the infection based on information related to the distribution width of the fluorescence intensity of the neutrophil group. The information related to the severity of the organ disorder caused by the infection is, for example, information related to the severity of the organ disorder of a subject suffering from the infection. Specifically, the information related to the severity of the organ disorder caused by the infection contains marker information indicating that the subject with the blood subject is suspected of having sepsis, and / or, the width of the estimated score indicating the severity of the organ disorder caused by the infection. The estimated score indicating the severity of the organ disorder caused by the infection is an estimated value of the SOFA score (hereinafter, also referred to as the "estimated SOFA score").

[0129] Specifically, the storage device 404 stores information related to the distribution width of the fluorescence intensity of the corresponding neutrophil population and a first conversion table for estimating the SOFA score. Fig. 9 An example of this first conversion table is shown. Fig. 9In the example, when the information related to the distribution width of the fluorescence intensity of the neutrophil group (NE-WY) is greater than 500 and less than 700, greater than 700 and less than 1000, greater than 1000 and less than 1300, and greater than 1300, the corresponding estimated SOFA scores are greater than 0 and less than 2, greater than 2 and less than 6, greater than 6 and less than 10, and greater than 10, respectively. In this way, in the first conversion table, the larger the distribution width of the fluorescence intensity of the neutrophil group, the larger the estimated score indicating the severity of the organ disorder caused by the infection. The first conversion table can be prepared by obtaining a regression equation for the data of sepsis patients and non-sepsis patients obtained by the method described in the above-mentioned experimental method. For example, Fig. 9 The first conversion table is from Figure 7 The regression equation shown in is derived.

[0130] The information processing unit 4 determines the width of the estimated score indicating the severity of the organ disorder caused by the infectious disease corresponding to the generated information related to the distribution width of the fluorescence intensity of the neutrophil population, based on the generated information related to the distribution width of the fluorescence intensity of the neutrophil population and the first conversion table by the main body 40. For example, when NE-WY is 600, the information processing unit 4 determines the width of the estimated score to be greater than 0 and less than 2.

[0131] The information processing unit 4 determines the flag information based on the determined width of the estimated score. The flag information is a text indicating that the subject with the blood sample is suspected of having sepsis, and is stored in the storage device 404. The text is, for example, "Sepsis?". Specifically, the information processing unit 4 acquires the flag information when, for example, the determined width of the estimated score is any one of 2 or more and less than 6, 6 or more and less than 10, and 10 or more.

[0132] The information processing unit 4 displays the analysis result of the blood sample on the output unit 42 . Fig. 10A This is a diagram showing an example of a screen displaying analysis results of a blood sample. Fig. 10B This is a diagram showing another example of a screen displaying analysis results of a blood sample. Fig. 10A and Fig. 10BThe analysis result screen 6 shown in is displayed by the main body 40 on the output unit 42. The analysis result screen 6 includes areas 61 to 64 in addition to the subject number and measurement date of the blood subject. Area 61 is an area that displays the number of neutrophils (NEUT), lymphocytes (LYMPH), monocytes (MONO), eosinophils (EO), and basophils (BASO) and their respective ratios to the number of white blood cells (WBC) together with the number of white blood cells (WBC). Area 62 is an area that displays data indicating an indicator of the distribution width of the fluorescence intensity of the neutrophil group. Among them, data of NE-WY is displayed in area 62. Area 63 is an area that displays information related to the severity of organ disorders caused by infections. Area 63 displays marker information indicating that the subject with the blood subject is suspected of having sepsis, and / or the width of an estimated score indicating the severity of organ disorders caused by infections. Fig. 10A In the example shown in , the text "Sepsis?" is displayed as flag information indicating that the subject is suspected of having sepsis. When the width of the estimated score is greater than 0 and less than 2, flag information on the severity of organ damage caused by the infection is not displayed. Fig. 10B In the example shown in FIG. , the width of the estimated SOFA score indicating the severity of organ failure caused by an infectious disease is shown (in Fig. 10B In the example of , 2 or more and less than 6). Fig. 10A and Fig. 10B In the example shown in , the width of the marker information and the estimated score may be displayed separately in area 63, or both may be displayed. Area 64 displays a scattergram generated by the information processing unit 4. This scattergram is a two-dimensional scattergram with the horizontal axis being the side scattered light intensity and the vertical axis being the fluorescence intensity.

[0133] As described above, the information processing unit 4 functions as an information processing unit that generates information related to the distribution width of the fluorescence intensity of the neutrophil group contained in the measurement sample based on the measured fluorescence signal, an information processing unit that determines information related to the severity of the organ disorder caused by the infection based on the information related to the distribution width of the fluorescence intensity of the neutrophil group, and an output unit that outputs information related to the severity of the organ disorder caused by the infection.

[0134] Next, the operation of the information processing unit 4 will be described. Fig.11 This is an example of an operation flow chart of the information processing unit, assuming that the information processing unit 4 controls the transport unit 3 to supply a blood sample to the measurement unit 2 .

[0135] like Fig.11As shown, the information processing unit 4 is controlled by the CPU 401 to perform measurement processing control (S1). The measurement processing control is a control that causes the measurement unit 2 to perform measurement processing, including modulation processing control and detection processing control. First, the information processing unit 4 performs modulation processing control (S1a). Specifically, the information processing unit 4 sends a first control signal to the measurement unit 2 to cause the measurement unit 2 to perform modulation processing. The modulation processing, as described above, is a process for preparing a measurement sample by mixing the blood sample, the first reagent, and the second reagent, and heating the mixture by the heater H in order to hemolyze the red blood cells contained in the blood sample and fluorescently stain the nucleic acid of the white blood cells.

[0136] Next, the information processing unit 4 performs detection processing control by the CPU 401 (S1b). Specifically, the information processing unit 4 sends a second control signal to the measuring unit 2 to make the measuring unit 2 perform detection processing. The detection processing is a detection processing of blood cells based on the measurement sample as described above. Specifically, the measuring unit 2, based on the second control signal, causes the measurement sample to flow to the flow cell D1 to generate a liquid flow, and the optical detector D measures the forward scattered light signal, the side scattered light signal, and the fluorescence signal of each blood cell. The measuring unit 2 sends the subject number corresponding to each measured signal to the information processing unit 4.

[0137] The information processing unit 4 stores the forward scattered light signal, the side scattered light signal, and the fluorescence signal of each blood cell received from the measuring unit 2 in association with the subject number received from the measuring unit 2 in the storage device 404. The information processing unit 4 generates a scattergram based on the received side scattered light signal and fluorescence signal by the CPU 401 (S2), and performs fractionation processing on the generated scattergram (S3). The scattergram generation processing and the segmentation processing are performed by using Figure 5A and Figure 5B The method described is carried out.

[0138] The information processing unit 4 counts the number of white blood cells (WBC) contained in the measurement sample and the number of blood cells contained in clusters A11 to A14 by CPU 401, and calculates the ratio of blood cells in each cluster to white blood cells (S4). The information processing unit 4 stores the counted number of white blood cells and the number of blood cells in each cluster and the calculated ratios in the storage device 404 in association with the subject number by CPU 401 (S4).

[0139] The information processing unit 4 generates information related to the distribution width of the fluorescence intensity of the neutrophil population by the CPU 401 (S5). Specifically, the information related to the distribution width of the fluorescence intensity of the neutrophil population is NE-WY.

[0140] Next, the information processing unit 4 determines the information related to the severity of the organ disorder caused by the infection based on the information related to the distribution width of the fluorescence intensity of the neutrophil group by the CPU 401 (S6). Specifically, the information processing unit 4 determines the width of the estimated SOFA score indicating the severity of the organ disorder caused by the infection corresponding to the NE-WY generated in S5 by referring to the first conversion table stored in the storage device 404 by the CPU 401. For example, when the value of NE-WY generated in step S5 is 500 or more and less than 700, the information processing unit 4 determines that the width of the estimated SOFA score is 0 or more and less than 2, and when the value of NE-WY is 700 or more and less than 1000, the width of the estimated SOFA score is 2 or more and less than 6. At this time, the information processing unit 4 reads the flag information from the storage device 404 when the width of the estimated SOFA score determined by the CPU 401 is any one of 2 or more and less than 6, 6 or more and less than 10, and 10 or more. Furthermore, the information processing unit 4 generates the analysis result screen 6 of the blood test body containing the information related to the severity of the organ disorder caused by the infection (in this embodiment, the width of the estimated SOFA score) by the CPU 401, and outputs (displays) it by the output unit 42 (S7). Furthermore, in the above-mentioned embodiment, the fluorescence signal of the measuring unit 2 as a waveform signal is sent to the information processing unit 4, and the information processing unit 4 obtains the fluorescence intensity based on the fluorescence signal, or the measuring unit 2 obtains the fluorescence intensity as a fluorescence signal and sends it to the information processing unit 4. In addition, the fluorescence intensity is a statistical value such as the peak value of the fluorescence signal of each blood cell or the average value of the fluorescence signal. In addition, the information related to the severity of the organ disorder caused by the infection can be output as text data or as image data.

[0141] [1-3. Functions and Effects of First Embodiment]

[0142] (1) The blood cell analyzer 1 of the present embodiment includes: a measuring unit (measuring unit 2) for irradiating light onto a measurement sample containing a blood sample and a fluorescent dye for staining white blood cells contained in the blood sample, detecting the fluorescence of the white blood cells contained in the measurement sample irradiated with light, and measuring a fluorescence signal based on the detected fluorescence; an information processing unit (main body 40) for generating information related to the distribution width of the fluorescence intensity of a neutrophil group contained in the measurement sample based on the measured fluorescence signal; determining information related to the severity of an organ disorder caused by an infectious disease based on the information related to the distribution width of the fluorescence intensity of the neutrophil group; and an output unit 42 for outputting information related to the severity of the organ disorder caused by the infectious disease.

[0143] Thus, a subject analysis method and a subject analysis device that support rapid evaluation of the severity of organ damage caused by an infection and that can output information highly correlated with a score indicating the severity of organ damage caused by an infection can be provided.

[0144] (2) Information related to the severity of organ damage caused by an infectious disease is set to include flag information indicating that the subject of the blood test sample is suspected of having sepsis. This can support the doctor's diagnosis that the subject has sepsis.

[0145] (3) The information related to the severity of organ disorders caused by the infection is set to include a width of an estimated score indicating the severity of organ disorders caused by the infection. This can support the determination of a treatment policy for the subject by a physician. For example, the larger the value included in the width of the estimated score, the more it can support the physician's determination of the intention to implement more diligent treatment.

[0146] (4) Information related to the distribution width of the fluorescence intensity of the neutrophil group is an index indicating the distribution width of the fluorescence intensity of the neutrophil group, and the larger this index is, the larger the estimated score indicating the severity of the organ failure caused by the infection becomes. Therefore, the larger the value of the estimated score is, the more it can support the physician to implement more diligent treatment.

[0147] (5) The blood cell analyzer 1 of the present embodiment is further provided with a preparation unit (sample preparation unit 25) for preparing a measurement sample. This makes it easy to analyze the leukocytes contained in the blood sample.

[0148] (6) The measurement section (measurement unit 2) includes a flow cell D1 through which the measurement sample flows, and light is irradiated to the measurement sample flowing through the flow cell D1. In this way, scattered light signals and fluorescence signals for each blood cell can be obtained.

[0149] [2. Second embodiment]

[0150] A blood cell analyzer according to a second embodiment will be described. Regarding the second embodiment, only portions different from the first embodiment will be described, and description of portions identical to the first embodiment will be omitted.

[0151] The method of obtaining the information related to the severity of organ damage caused by an infectious disease in the second embodiment is different from that in the first embodiment. In addition, the information related to the severity of organ damage caused by an infectious disease in the second embodiment includes an estimated score indicating the severity of the organ damage caused by an infectious disease. This estimated score is an estimated SOFA score.

[0152] Specifically, the information processing unit 4 generates the estimated SOFA score from the regression equation that corresponds the distribution width related information (NE-WY) of the fluorescence intensity of the neutrophil group and the estimated SOFA score by the main body 40. That is, in the second embodiment, a computer program including the regression equation is stored in the storage device 404 instead of the first conversion table of the first embodiment, and the information processing unit 4 generates the estimated SOFA score corresponding to NE-WY by executing the computer program including the regression equation by the main body 40. This regression equation is, for example, Figure 7 y=0.013x-7.1 explained in . Therefore, since the regression equation is a linear equation with a positive slope, the larger the NE-WY is, the larger the estimated score indicating the severity of organ failure caused by the infection becomes. The regression equation can also be calculated from data of a sufficient number of sepsis patients and non-sepsis patients with sufficient reliability.

[0153] Fig.12 FIG. 2 is a diagram showing an example of a screen showing analysis results of a blood sample according to the second embodiment. Fig.12 As shown, the information processing unit 4 displays the generated estimated SOFA score in the area 63 of the analysis result screen 6. Fig.12 In the example shown in , the estimated SOFA score is shown to be 2.

[0154] In this way, even if the information related to the severity of organ damage caused by an infectious disease includes an estimated score indicating the severity of organ damage caused by an infectious disease, a subject analysis method and a subject analysis device that support rapid evaluation of the severity of organ damage caused by an infectious disease can be provided, which is a subject analysis method and a subject analysis device that can output information highly related to the score indicating the severity of organ damage caused by an infectious disease. For example, the larger the value of the estimated score, the more it can support the doctor's determination of the intention to implement more diligent treatment.

[0155] In addition, the information related to the distribution width of the fluorescence intensity of the neutrophil group is an index indicating the distribution width of the fluorescence intensity of the neutrophil group, and the larger this index is, the larger the estimated score indicating the severity of the organ failure caused by the infection becomes. Therefore, the larger the value of the estimated score is, the more it can support the physician to implement more diligent treatment.

[0156] [3. Third embodiment]

[0157] A blood cell analyzer according to a third embodiment will be described. Regarding the third embodiment, only portions different from the first embodiment will be described, and description of portions identical to the first embodiment will be omitted.

[0158] The information related to the severity of organ damage caused by an infection according to the third embodiment is different from that of the first embodiment. That is, the information related to the severity of organ damage caused by an infection according to the third embodiment is flag information indicating that the subject with the blood sample is suspected of having sepsis in multiple stages.

[0159] Fig.13 This is an example of the second conversion table that corresponds to NE-WY and the probability of sepsis in multiple stages. Fig.13 As shown, in the second conversion table, when NE-WY is 500 or more and less than 700, 700 or more and less than 1300, or 1300 or more, the possibility of sepsis corresponds to low, medium, and high, respectively. In the third embodiment, a second conversion table is stored in the storage device 404 instead of the first conversion table of the first embodiment, and the information processing unit 4 determines the possibility of sepsis of the subject based on the generated NE-WY and the second conversion table by the main body 40.

[0160] Fig.14 FIG. 2 is a diagram showing an example of a screen showing analysis results of a blood sample according to the third embodiment. Fig.14 As shown, the information processing unit 4 displays the acquired likelihood of sepsis as flag information in the area 63 of the analysis result screen 6. Fig.14 In the example shown in , the possibility of sepsis is high.

[0161] Even if the information related to the severity of organ failure caused by an infection is flag information indicating the degree of possibility of sepsis, a subject analysis method and a subject analysis device that support rapid evaluation of the severity of organ failure caused by an infection can be provided, which is a subject analysis method and a subject analysis device that can output information highly correlated with the score indicating the severity of organ failure caused by an infection. For example, the higher the stage indicating the severity, the more the doctor can be supported to perform more diligent treatment.

[0162] In the present embodiment, the possibility of sepsis is set to three stages as flag information, but may be set to two stages, four stages, or five or more stages. In addition, the possibility of sepsis may be expressed as a probability.

[0163] [4. Fourth embodiment]

[0164] A blood cell analyzer according to a fourth embodiment will be described. Regarding the fourth embodiment, only portions different from the first embodiment will be described, and description of portions identical to the first embodiment will be omitted.

[0165] The information related to the severity of organ damage caused by an infectious disease according to the fourth embodiment is different from that of the first embodiment. That is, the information related to the severity of organ damage caused by an infectious disease according to the fourth embodiment includes flag information indicating that the subject having the blood test sample is suspected of having sepsis, and information indicating whether the estimated value of the difference between the SOFA score at the initial stage of hospitalization and the SOFA score when healthy is above a threshold.

[0166] Specifically, the storage device 404 stores a threshold value instead of the first conversion table. In Sepsis-3, since sepsis is defined when an infection and a sudden increase of 2 or more points in the SOFA score are observed, the threshold value can be set to, for example, a value of information related to the distribution width of the fluorescence intensity of the neutrophil group corresponding to an estimated value of the difference between the SOFA score at the beginning of hospitalization and the SOFA score when healthy (NE-WY). However, the threshold value can also be set as appropriate.

[0167] When the information processing unit 4 compares the generated information on the distribution width of the fluorescence intensity of the neutrophil group (NE-WY) with the threshold value read from the storage device 404 by the main body 40 and determines that NE-WY is above the threshold value, the information processing unit 4 displays flag information indicating that the estimated value of the difference between the SOFA score at the beginning of hospitalization and the SOFA score when healthy is above the threshold value in the area 63 of the analysis result screen 6 displayed on the output unit 42. When the information processing unit 4 determines that NE-WY is below the threshold value by the main body 40, the information processing unit 4 displays flag information indicating that the estimated value of the difference between the SOFA score at the beginning of hospitalization and the SOFA score when healthy is below the threshold value in the area 63 of the analysis result screen 6 displayed on the output unit 42. Furthermore, when NE-WY is below the threshold value, the flag information may be non-displayed.

[0168] Fig.15 This is a diagram showing an example of a screen displaying analysis results of a blood sample according to the fourth embodiment. Fig.15 This is the analysis result screen 6 when NE-WY is above the threshold. Fig.15 As shown, in area 63, "estimated ΔSOFA score ≥ 2" is displayed, indicating that the estimated value of the difference between the SOFA score at the initial stage of hospitalization and the SOFA score during health is 2 or more.

[0169] For the threshold setting method, use Fig.16 illustrate.

[0170] Fig.16 This is a graph showing the correlation between the distribution width information (NE-WY) of the fluorescence intensity of the neutrophil group of hospitalized patients on each day and the difference between the SOFA score on each day and the SOFA score in healthy patients. Fig.16The leftmost graph, the center graph, and the rightmost graph are graphs showing the correlation between NE-WY on hospital day 0, hospital day 1, and hospital day 3 and the difference between the SOFA score on each day and the SOFA score when healthy. The white dots in each graph are data of sepsis patients, and the black dots are data of non-sepsis patients. Figure 8 The same patient is used for each patient. The calculation method of the information related to the distribution width of the fluorescence intensity of the neutrophil group of the hospitalized patients on each day (NE-WY), and the blood cell analyzer used in the calculation are the same as those described above. The SOFA score when healthy is the SOFA score of each patient at the time of discharge. The difference between the SOFA score of each day and the SOFA score when healthy is calculated by subtracting the SOFA score when healthy from the SOFA score of each patient on each day.

[0171] The Spearman's rank-order correlation coefficient r, which indicates the correlation between NE-WY and the difference between the SOFA score and the healthy SOFA score, is 0.67 on the first day of hospitalization (day 0), 0.53 on the first day of hospitalization, and 0.23 on the third day of hospitalization. The highest correlation was confirmed on the first day of hospitalization (day 0), and then a high correlation was confirmed on the first day of hospitalization. In addition, it was confirmed that the correlation decreased every day from the beginning of hospitalization. The reason for the decrease in daily correlation is believed to be that the infection was restrained according to the treatment due to the administration of medication to suppress infection from the first day of hospitalization. In addition, the inventors have found that Fig.16 The regression equations were calculated for the data of sepsis patients and non-septic patients on each day shown in . The regression equations for the first day of hospitalization (day 0), the first day of hospitalization, and the third day of hospitalization were y=0.013x-7.3, y=0.015x-8.5, and y=0.022x-12.1, respectively. However, the x-axis was set to NE-WY, and the y-axis was set to the difference between the SOFA score of each day and the SOFA score when healthy.

[0172] The threshold can be obtained from Fig.16 The regression equation corresponding to the index showing the distribution width of the fluorescence intensity of the neutrophil group and the difference between the SOFA score and the healthy SOFA score is obtained. For example, NE-WY corresponding to the difference of 2 can be set as the threshold. In Sepsis-3, sepsis is defined when an infection and a sudden increase of 2 points or more in the SOFA score are seen.

[0173] The threshold value can be obtained for each of the first day of hospitalization (day 0), the first day of hospitalization, and the third day of hospitalization, and stored in the storage device 404. The information processing unit 4 can also be configured to select a threshold value corresponding to the day of hospitalization of the subject and obtain flag information. For example, when the subject is on the first day of hospitalization, the information processing unit 4 receives input of information that "the subject is on the first day of hospitalization" from the user's input unit 41 or the host computer 5, reads the threshold value for the first day of hospitalization from the storage device 404, compares the threshold value with NE-WY, and displays flag information of "estimated ΔSOFA score ≥ 2" or "estimated ΔSOFA score < 2" in the area 63 of the analysis result screen 6 displayed on the output unit 42. Furthermore, the threshold value can also be used independently of the number of days from the first day of hospitalization, and a common threshold value can be used.

[0174] As a first variation of the present embodiment, a regression equation may be prepared for each number of days since hospitalization, and a computer program including the regression equation may be stored in the storage device 404. In this case, the information processing unit 4 executes the computer program including the regression equation corresponding to the number of days since hospitalization of the subject by the main body 40, and determines flag information indicating whether the estimated value of the difference between the SOFA score at the initial stage of hospitalization and the SOFA score during health is above a threshold value. The number of days since hospitalization of the subject is input from the input unit 41 or the host computer 5.

[0175] As a second variation of the present embodiment, a computer program including a common regression equation not depending on the number of days since hospitalization may be stored in the storage device 404. In this case, the information processing unit 4 executes the computer program through the main body 40 to determine flag information indicating whether the estimated value of the difference between the SOFA score at the initial stage of hospitalization and the SOFA score during health is above a threshold value.

[0176] As described above, in this embodiment, the flag information is set to include information indicating whether the estimated value of the difference between the SOFA score at the initial stage of hospitalization and the SOFA score when healthy is above the threshold. Thus, a subject analysis method and a subject analysis device that support rapid evaluation of the severity of organ disorders caused by infectious diseases can be provided, and a subject analysis method and a subject analysis device that can output information highly correlated with the score indicating the severity of organ disorders caused by infectious diseases can be provided. Specifically, while supporting the physician's diagnosis that the subject has sepsis, it is possible to support the physician's determination of a treatment policy for the subject.

[0177] [5. Other embodiments]

[0178] The above-described embodiments are examples for explaining the present invention, and the present invention is not limited to these embodiments. The present invention can be implemented in various forms as long as it does not deviate from the gist. For example, any combination of two or more of the first to fourth embodiments and modified examples is also included in the present invention. In addition, in the above-described processing or action, as long as there is no contradiction in the processing or action of a certain step using data that should not be available in the step, the order of the processing or action and the execution subject can be freely changed.

[0179]

Explanation of symbols

[0180] 1: Blood cell analysis device

[0181] 2: Measurement unit

[0182] 21: Hands

[0183] 22: Test subject container installation section

[0184] 23: Barcode unit

[0185] 24: Subject suction part

[0186] 25: Sample preparation department

[0187] 26: Inspection Department

[0188] D: Optical detector

[0189] D1: Flow cell

[0190] D2: Sheath flow system

[0191] D3: Beam point forming system

[0192] D31: Semiconductor Laser

[0193] D32: Collimating Tube Lens

[0194] D33: Condenser lens

[0195] D34: Beam Stopper

[0196] D4: Forward scattered light receiving system

[0197] D41: Front focusing lens

[0198] D42: Pinhole

[0199] D43: Photodiode

[0200] D5: Side scattered light receiving system

[0201] D51: Side focusing lens

[0202] D52: Dichroic Mirror

[0203] D53: Photodiode

[0204] D6: Fluorescence light receiving system

[0205] D61: Spectral filter

[0206] D62: Photodiode

[0207] D7: Signal processing circuit

[0208] 3: Transport unit

[0209] 4: Information processing unit

[0210] 40: Ontology

[0211] 401: CPU

[0212] 402: ROM

[0213] 403: RAM

[0214] 404: Storage Device

[0215] 404a: Program

[0216] 405: Read Device

[0217] 405a: Recording medium

[0218] 406: Image output interface

[0219] 407: Input and output interface

[0220] 408: Communication interface

[0221] 41: Input section

[0222] 42: Output section

[0223] 5: Main computer

Claims

1. A subject analysis method, comprising: a measurement step of irradiating light onto a measurement sample containing a blood sample and a fluorescent dye for staining leukocytes contained in the blood sample, detecting fluorescence of the leukocytes contained in the measurement sample irradiated with the light, and measuring a fluorescence signal based on the detected fluorescence, a step of generating information related to a distribution width of fluorescence intensity of a neutrophil population contained in the measurement sample based on the fluorescence signal, a step of determining information related to the severity of organ damage caused by the infection based on the information related to the distribution width of the fluorescence intensity of the neutrophil population, and an outputting step of outputting information related to the severity of the organ failure caused by the infection.

2. The subject analysis method according to claim 1, wherein the information related to the severity of the organ disorder caused by the infection includes flag information indicating the degree of suspicion of sepsis or the possibility of sepsis in the subject of the blood subject.

3. The subject analysis method according to claim 1, wherein the information related to the severity of the organ damage caused by the infection includes an estimated score indicating the severity of the organ damage caused by the infection. 4 . The subject analysis method according to claim 1 , wherein the information related to the severity of the organ damage caused by the infection includes a width of an estimated score indicating the severity of the organ damage caused by the infection.

5. The subject analysis method according to claim 3 or 4, wherein The information related to the distribution width of the fluorescence intensity of the neutrophil population indicates the distribution width of the fluorescence intensity of the neutrophil population. The larger the distribution width is, the larger the estimated score indicating the severity of the organ damage caused by the infection becomes. 6 . The subject analysis method according to claim 3 , wherein the estimated score indicating the severity of the organ failure caused by the infection is an estimated value of the SOFA score.

7. The subject analysis method according to claim 6, wherein the information related to the severity of the organ disorder caused by the infection includes information indicating whether an estimated value of the difference between the SOFA score at the initial stage of hospitalization and the SOFA score when healthy is greater than a threshold value.

8. The subject analysis method according to claim 1, wherein the measurement step further comprises a preparation step of preparing the measurement sample.

9. The subject analysis method according to claim 1, wherein The measurement step further includes a flow step of causing the measurement sample to flow into a flow cell. In the measurement step, the measurement sample flowing through the flow cell is irradiated with the light.

10. A subject analysis device, comprising: a measuring unit that irradiates light onto a measuring sample containing a blood sample and a fluorescent dye that stains leukocytes contained in the blood sample, detects fluorescence of leukocytes contained in the measuring sample irradiated with the light, and measures a fluorescence signal based on the detected fluorescence, an information processing unit that generates information related to a distribution width of fluorescence intensity of a neutrophil group contained in the measurement sample based on the fluorescence signal, determines information related to the severity of an organ disorder caused by an infectious disease based on the information related to the distribution width of fluorescence intensity of the neutrophil group, and An output unit outputs information related to the severity of the organ failure caused by the infectious disease.

11. The subject analysis device according to claim 10, wherein the information related to the severity of the organ disorder caused by the infection includes flag information indicating the degree of suspicion of sepsis or possibility of sepsis in the subject of the blood subject.

12. The subject analysis device according to claim 10, wherein the information related to the severity of the organ damage caused by the infection includes an estimated score indicating the severity of the organ damage caused by the infection.

13. The subject analysis device according to claim 10, wherein the information related to the severity of the organ damage caused by the infection includes a width of an estimated score indicating the severity of the organ damage caused by the infection.

14. The subject analysis device according to claim 12 or 13, wherein The information related to the distribution width of the fluorescence intensity of the neutrophil population indicates the distribution width of the fluorescence intensity of the neutrophil population. The larger the distribution width is, the larger the estimated score indicating the severity of the organ damage caused by the infection becomes. 15 . The subject analysis device according to claim 14 , wherein the estimated score indicating the severity of the organ failure caused by the infection is an estimated value of the SOFA score.

16. The subject analysis device according to claim 12 or 13, wherein the information related to the severity of the organ damage caused by the infection includes information indicating whether an estimated value of the difference between the SOFA score at the initial stage of hospitalization and the SOFA score when healthy is greater than a threshold value.

17. The subject analyzing device according to claim 10, wherein the measuring section further comprises a preparing section for preparing the measurement sample.

18. The subject analysis device according to claim 10, wherein The measuring section includes a flow cell through which the measuring sample flows. The measurement sample flowing through the flow cell is irradiated with the light.

19. The subject analysis device according to claim 10, comprising at least one of the following configurations [11] to [18]: [11] the information related to the severity of organ damage caused by the infection includes marker information indicating that the subject of the blood test sample is suspected of having sepsis, [12] the information related to the severity of the organ disorder caused by the infectious disease includes an estimated score indicating the severity of the organ disorder caused by the infectious disease, [13] the information related to the severity of the organ disorder caused by the infectious disease includes a width of an estimated score indicating the severity of the organ disorder caused by the infectious disease, [14] the information related to the distribution width of the fluorescence intensity of the neutrophil population indicates the distribution width of the fluorescence intensity of the neutrophil population, and the larger the distribution width, the larger the estimated score indicating the severity of the organ damage caused by the infection becomes, [15] The estimated score indicating the severity of the organ failure caused by the infection is an estimated value of the SOFA score, [16] The flag information includes information indicating whether the estimated value of the difference between the SOFA score at the initial stage of hospitalization and the SOFA score during health is above a threshold value, [17] The measuring unit further includes a modulation unit for modulating the measuring sample, [18] The measurement unit includes a flow cell through which the measurement sample flows, and the measurement sample flowing through the flow cell is irradiated with the light.

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