Blood analysis device

Abstract

The present invention provides an analysis device for easily judging whether to provide an iron supplement to an iron deficiency anemia patient, comprising: a first sample preparation unit (5A) for preparing a first measurement sample for measuring red blood cells infected with malaria from a blood sample; a second sample preparation unit (5B) for preparing a second measurement sample for measuring the autofluorescence of red blood cells from a blood sample; a light source unit (62) for irradiating light to the first measurement sample and the second measurement sample; a detector (63, 64) for detecting the fluorescence and scattered light generated by the first measurement sample irradiated with light, and detecting the autofluorescence generated by the second measurement sample irradiated with light; and an information processing unit (3) for generating information related to malaria infection based on the fluorescence and scattered light detected from the first measurement sample, and generating information related to iron deficiency anemia based on the autofluorescence of red blood cells detected from the second measurement sample.

Description

Technical Field

[0001] The present invention relates to a blood analysis device. Background Art

[0002] Malaria parasites use heme iron as food to survive and reproduce. Therefore, when a patient with iron deficiency anemia is infected with malaria, if the doctor does not notice the malaria infection and gives the patient with iron deficiency anemia iron supplements, the malarial symptoms will worsen. Therefore, in areas where malaria has become endemic, the World Health Organization (WHO) recommends providing iron supplements to patients with iron deficiency anemia together with malaria diagnosis (for example, refer to non-patent document 1). For example, malaria diagnosis is performed by detecting DNA aptamers that specifically bind to pLDH (plasmodium lactate dehydrogenase: plasmodium lactate dehydrogenase) (for example, refer to patent document 1). The DNA aptamer amplified by the polymerase chain reaction (PCR) is fluorescently labeled, and the excited fluorescence is detected, thereby detecting the DNA aptamer. On the other hand, for example, iron deficiency anemia is diagnosed by detecting antibodies that bind to repulsive guidance molecules (Repulsive Guidance Molecules) (for example, refer to patent document 2). Antibody detection is performed by enzyme-linked immunosorbent assay (ELISA).

[0003] Prior art literature

[0004] Non-patent literature

[0005] Non-patent literature 1: GUIDELINE: DAILY IRON SUPPLEMENTATION IN INFANTS AND CHILDREN, World Health Organization, 2016;

[0006] Patent document 1: Japanese Patent Application Publication No. 2012-254074;

[0007] Patent document 2: Japanese Patent Application Publication No. 2015-502753. Summary of the invention

[0008] Technical problem to be solved by the invention

[0009] As described above, the diagnosis of malaria requires amplification of DNA aptamers by polymerase chain reaction and detection of excitation fluorescence. On the other hand, the diagnosis of iron deficiency anemia requires binding to specific antibodies and detection of antibodies by enzyme-linked immunosorbent assay. Therefore, in order to determine whether to provide iron supplements to patients with iron deficiency anemia, it is necessary to prepare an analysis device for malaria diagnosis and an analysis device for iron deficiency anemia diagnosis, collect samples for each analysis device, and perform measurements.

[0010] Therefore, one of the objects of the present invention is to provide an analysis device that can easily perform measurement for determining whether to provide an iron supplement to a patient with iron deficiency anemia.

[0011] Technical means to solve technical problems

[0012] A blood analyzer is provided by the technical solution of the present invention, comprising: a first sample preparation unit for preparing a first measurement sample for measuring red blood cells infected with malaria from a blood sample; a second sample preparation unit for preparing a second measurement sample for measuring the autofluorescence of red blood cells from a blood sample; a light source unit for irradiating light to the first measurement sample and the second measurement sample; a detection unit for detecting fluorescence and scattered light generated by the first measurement sample irradiated with light, and detecting autofluorescence generated by the second measurement sample irradiated with light; and an information processing unit for generating information related to malaria infection based on the fluorescence and scattered light detected from the first measurement sample, and generating information related to iron deficiency anemia based on the autofluorescence detected from the second measurement sample.

[0013] Furthermore, in the above-mentioned blood analysis device of the present invention, the information processing unit outputs display information for simultaneously outputting information related to malaria infection and information related to iron deficiency anemia to a display device, the information related to malaria infection including a comparison result of the number of red blood cells infected with malaria or the ratio of the number of red blood cells infected with malaria to the number of red blood cells with a threshold value for judging the possibility of malaria infection, and the information related to iron deficiency anemia including a comparison result of the number of red blood cells emitting autofluorescence or the ratio of the number of red blood cells emitting autofluorescence to the number of red blood cells with a threshold value for judging the possibility of iron deficiency anemia; when the comparison result with the threshold value for judging the possibility of malaria infection indicates the possibility of malaria infection, and the comparison result with the threshold value for judging the possibility of iron deficiency anemia indicates the possibility of iron deficiency anemia, the information processing unit generates a mark for warning attention.

[0014] Effects of the Invention

[0015] According to the present invention, information for determining whether to provide an iron supplement to an iron-deficiency anemia patient is generated by one blood analyzer, so that measurement for determining whether to provide an iron supplement to an iron-deficiency anemia patient can be easily performed. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 is a schematic diagram of the structure of a blood analysis device according to an embodiment;

[0017] Figure 2 is a schematic diagram of the structure of a blood analysis device according to an embodiment;

[0018] Figure 3 A block diagram showing a structure of an information processing unit of a blood analyzer according to an embodiment of the present invention;

[0019] Figure 4 is a flowchart of the operation of the blood analysis device involved in the embodiment;

[0020] Figure 5 A flowchart of the procedure of the first measurement sample preparation process according to the embodiment;

[0021] Figure 6 This is a flowchart of the procedure of the second measurement sample preparation process according to the embodiment;

[0022] Figure 7 This is a flowchart of the procedure of the first measurement data analysis process according to the embodiment;

[0023] Figure 8 A schematic diagram of the appearance regions of each particle group of white blood cells, platelet aggregation, normal red blood cells and malaria-infected red blood cells in a scatter diagram in which the vertical axis is the forward scattered light intensity and the horizontal axis is the fluorescence intensity according to an embodiment;

[0024] Fig. 9 A flowchart of the procedure of the malaria type determination process according to the embodiment;

[0025] Fig.10 This is an illustration of the life cycle of Plasmodium;

[0026] Fig.11 A schematic diagram of the appearance areas of each particle group of white blood cells, platelet aggregation, normal red blood cells, single ring bodies, multiple ring bodies, trophozoites, schizonts and gametocytes in a scatter diagram in which the vertical axis is the forward scattered light intensity and the horizontal axis is the fluorescence intensity according to the embodiment;

[0027] Fig.12 This is a scatter plot of a blood sample infected with Plasmodium;

[0028] Fig.13 This is a scatter plot of a blood sample infected with Plasmodium;

[0029] Fig.14 This is a scatter plot of a blood sample infected with Plasmodium;

[0030] Fig.15 This is a flowchart of the procedure of the second measurement data analysis process according to the embodiment;

[0031] Fig.16A A scattergram showing measurement results of a normal sample according to an example of an embodiment;

[0032] Fig. 16B A scattergram showing measurement results of a blood sample collected from a patient with iron deficiency anemia according to an example of an embodiment;

[0033] Fig. 16C A scattergram showing measurement results of a blood sample collected from an α-thalassemia patient according to an example of an embodiment;

[0034] Fig.16D A scattergram showing measurement results of a blood sample collected from a β-thalassemia patient according to an example of an embodiment;

[0035] Fig.17 A schematic diagram showing an example of displaying analysis results according to an embodiment of the present invention;

[0036] Fig.18 A diagram showing the relationship between a combination of a malaria type and stage determination mark and an iron deficiency anemia determination mark and a displayed message;

[0037] Fig.19 A graph showing the temporal changes in the ratio of the number of malaria-infected erythrocytes, the number of autofluorescent erythrocytes, and the number of erythrocytes in the peripheral blood of a mouse infected with malaria according to an embodiment;

[0038] Fig. 20 This is a scatter plot of the measurement sample prepared from the peripheral blood of mice infected with malaria according to the embodiment;

[0039] Fig.21 The fluorescence microscopy images of the peripheral blood of mice infected with malaria involved in the example. DETAILED DESCRIPTION

[0040] The following is an explanation of the embodiments of the present invention with reference to the accompanying drawings. In the description of the following drawings, the same or similar parts are represented by the same or similar symbols. However, the accompanying drawings are schematic diagrams. Therefore, specific dimensions, etc. should be judged by comparing with the following description. In addition, the drawings also include parts with different dimensional relationships and proportions.

[0041] In this embodiment, a blood analysis device is described, which determines whether a person is infected with malaria based on the fluorescence intensity and scattered light intensity generated when a first measurement sample prepared from a blood sample and a reagent is irradiated with light, and determines whether the person has iron deficiency anemia based on the autofluorescence intensity of red blood cells contained in a second measurement sample prepared from the blood sample.

[0042] <Structure of blood analyzer>

[0043] like Figure 1As shown, the blood analyzer according to the embodiment includes a measuring unit 2 and an information processing unit 3. The measuring unit 2 sucks a blood sample and prepares a first and a second measurement sample from the blood sample, and optically measures the first and the second measurement sample, respectively. The information processing unit 3 processes the first measurement data derived from the first measurement sample and the second measurement data derived from the second measurement sample obtained by the measurement of the measuring unit 2, and outputs the analysis result of the blood sample.

[0044] The measurement unit 2 includes a first sample preparation unit 5A that prepares a first measurement sample for measuring malaria-infected erythrocytes from a blood sample, and a second sample preparation unit 5B that prepares a second measurement sample for measuring autofluorescence of erythrocytes from a blood sample.

[0045] The measurement unit 2 further includes an aspirating unit 4. The aspirating unit 4 includes a pipette 20 and a quantitative unit 21, and aspirates the blood sample contained in the test tube via the pipette 20.

[0046] The first sample preparation unit 5A has a first reaction tank 54A and is connected to reagent containers 51, 52, and 53. The reagent container 51 contains a hemolytic agent that shrinks red blood cells. The reagent container 52 contains a nucleic acid staining reagent containing a dye that stains nucleic acids such as DNA. The reagent container 53 contains a diluent that does not contain a hemolytic agent or a dye.

[0047] The aspiration unit 4 moves the aspiration tube 20 to the top of the first reaction tank 54A and discharges the aspirated blood sample to the first reaction tank 54A. The hemolytic agent is sent from the reagent container 51 to the first reaction tank 54A. The nucleic acid staining reagent is sent from the reagent container 52 to the first reaction tank 54A. The blood sample, the hemolytic agent, and the nucleic acid staining reagent are mixed in the first reaction tank 54A. Further, a diluent is sent from the reagent container 53 to the first reaction tank 54A, and the reaction solution of the blood sample, the hemolytic agent, and the nucleic acid staining reagent is diluted by the diluent, thereby preparing the first measurement sample. The first measurement sample is used for the measurement of malaria-infected red blood cells.

[0048] The hemolytic agent contained in the reagent container 51 contains, for example, two surfactants with different hemolytic abilities. For example, the hemolytic agent contains 2.95 mmol / L of lauryltrimethyl lammonium chloride as a cationic surfactant and 1.11 mmol / L of stearyl trimethyl lammonium chloride as a cationic surfactant. Stearyl trimethyl lammonium chloride has a stronger hemolytic ability than lauryl trimethyl ammonium chloride. As long as the two surfactants have different hemolytic abilities, they are not limited to the above combination. In addition, the hemolytic agent also contains 2.90 mmol / L of PBC-44 as a nonionic surfactant, 20 mmol / L of ADA (adenosine deaminase), an appropriate amount of NaCl and 1 L of pure water. The pH of ADA is 6.1. In addition, the pH of the hemolytic agent is 5.0 or more and 7.0 or less. In order to shrink blood cells, the osmotic pressure of the hemolytic agent is set to, for example, 200 mOsm / kg·H 2 O or more and 300 mOsm / kg·H 2 O or less. An example of the hemolytic agent is Lysercell (Sysmex).

[0049] The nucleic acid staining reagent contained in the reagent container 52 contains, for example, a staining dye (Hoechst 34580). The concentration of the staining dye in the nucleic acid staining reagent is, for example, 0.3 μmol / L to 0.6 μmol / L, or 0.45 μmol / L. In addition, the concentration of the staining dye in the measurement sample is, for example, 0.15 μmol / L to 1.0 μmol / L. The chemical formula of Hoechst 34580 is as follows.

[0050]

Chemistry 1

[0051]

[0052] Alternatively, the first measurement sample may be prepared by mixing a single reagent containing a hemolytic agent that shrinks red blood cells and a staining dye with a blood sample without separating the hemolytic agent and the nucleic acid staining reagent.

[0053] Buffer solutions such as cellpack DFL (Sysmex) can be cited as examples of the diluent containing no hemolytic agent and no dye contained in the reagent container 53. In the measurement of blood cells by flow cytometry, the diluent containing no hemolytic agent is also used as a sheath fluid.

[0054] The second sample preparation part 5B has a second reaction tank 54B, and is connected to the reagent container 53 .

[0055] The aspirating unit 4 moves the aspirating tube 20 to the upper side of the second reaction tank 54B and discharges the aspirated blood sample to the second reaction tank 54B. The diluent is sent from the reagent container 53 to the second reaction tank 54B. The blood sample and the diluent are mixed in the second reaction tank 54B to prepare a second measurement sample. The second measurement sample is used to measure the autofluorescence of red blood cells.

[0056] The structures of the first sample preparation unit 5A and the second sample preparation unit 5B are described in further detail. Figure 2 As shown, the aspiration unit 4 includes a pipette 20 for liquid to pass through the inside and a quantitative unit 21. The front end of the pipette 20 can penetrate (puncture) the sealing cap 100a of the sample container 100 that stores the sample. The pipette 20 can move in the vertical direction (Z direction) and can move to the first reaction tank 54A and the second reaction tank 54B. The quantitative unit 21 uses a syringe pump. In addition, for example, a diaphragm pump can also be used instead of the syringe pump. The quantitative unit 21 aspirates a certain amount of sample from the sample container 100 through the pipette 20 and discharges it. As a result, a certain amount of sample required for sample measurement is aspirated from the sample container 100, and the sample can be supplied to the first reaction tank 54A and the second reaction tank 54B.

[0057] A quantitative unit 22 and electromagnetic valves 23 and 24 are provided in the flow path between the reagent container 51 containing the hemolytic agent and the first reaction tank 54A. The quantitative unit 22 uses a syringe pump. In addition, for example, a diaphragm pump can be used instead of the syringe pump. The electromagnetic valves 23 and 24 open or close the flow path. The quantitative unit 22 and the electromagnetic valves 23 and 24 quantitatively deliver the hemolytic agent in the reagent container 51 to the first reaction tank 54A.

[0058] The reagent container 52 storing the reagent for nucleic acid staining is placed by the reagent container holder 25. The reagent container holder 25 is provided with a pipette 26 for aspirating the reagent for nucleic acid staining in the reagent container 52, and a pipette lifting mechanism 27 for lifting and lowering the pipette 26. The front end of the pipette 26 can penetrate (puncture) the sealing member of the reagent container 52. The cover 28 is connected to the pipette lifting mechanism 27. When the pipette lifting mechanism 27 is lowered and the pipette 26 penetrates (punctures) the sealing member of the reagent container 52, the cover 28 also descends and covers the reagent container 52. When the pipette lifting mechanism 27 is raised, the cover 28 is also raised, so that the reagent container 52 can be removed from the outside.

[0059] A bubble sensor 29 for detecting bubbles in the liquid is provided in the flow path between the pipette 26 and the first reaction tank 54A. In addition, a quantitative unit 30 and electromagnetic valves 31, 32 are provided in the flow path between the pipette 26 and the first reaction tank 54A. The quantitative unit 30 uses a syringe pump. In addition, for example, a diaphragm pump can also be used instead of the syringe pump. The electromagnetic valves 31, 32 open or close the flow path. The quantitative unit 30 and the electromagnetic valves 31, 32 quantitatively deliver the nucleic acid staining reagent in the reagent container 52 to the first reaction tank 54A.

[0060] A quantitative unit 33 and electromagnetic valves 34, 35 are provided in the flow path between the reagent container 53 for storing the diluent and the first reaction tank 54A. The quantitative unit 33 uses a syringe pump. In addition, for example, a diaphragm pump can also be used instead of the syringe pump. The electromagnetic valves 34, 35 open or close the flow path. The quantitative unit 33 and the electromagnetic valves 34, 35 quantitatively send the diluent in the reagent container 53 to the first reaction tank 54A. The first reaction tank 54A is connected to a waste liquid chamber 36 for storing unnecessary solutions. A solenoid valve 37 for opening or closing the flow path is provided between the first reaction tank 54A and the waste liquid chamber 36.

[0061] A heater 55A for heating the liquid in the first reaction tank 54A is provided adjacent to the first reaction tank 54A. In addition, the first reaction tank 54A is connected to a pump 56A for supplying air into the first reaction tank 54A in order to stir the liquid in the first reaction tank 54A.

[0062] A quantitative unit 38 and electromagnetic valves 39 and 40 are provided in the flow path between the reagent container 53 for storing the diluent and the second reaction tank 54B. The quantitative unit 38 uses a syringe pump. In addition, for example, a diaphragm pump can also be used instead of the syringe pump. The electromagnetic valves 39 and 40 open or close the flow path. The quantitative unit 38 and the electromagnetic valves 39 and 40 quantitatively send the diluent in the reagent container 53 to the second reaction tank 54B. The second reaction tank 54B is connected to the waste liquid chamber 41 for storing unnecessary solutions. A solenoid valve 42 is provided between the second reaction tank 54B and the waste liquid chamber 41, and the solenoid valve 42 switches the flow path between the flow path from the second reaction tank 54B to the waste liquid chamber 41 and the flow path from the second reaction tank 54B to the second detection device 7.

[0063] A heater 55B for heating the liquid in the second reaction tank 54B is provided adjacent to the second reaction tank 54B. In addition, the second reaction tank 54B is connected to a pump 56B for supplying air into the second reaction tank 54B in order to stir the liquid in the second reaction tank 54B.

[0064] Figure 1The measurement unit 2 shown in the figure further includes an optical detection device 6, which measures white blood cells, malarial infected red blood cells and the autofluorescence of red blood cells by flow cytometry. The optical detection device 6 includes: a flow chamber 61 for each of the first and second measurement samples to flow; a light source unit 62 for irradiating light to the first and second measurement samples flowing in the flow chamber 61; and a detection unit 6A for detecting the intensity of the fluorescence and the intensity of the scattered light generated by each of the first and second measurement samples irradiated with light. The detection unit 6A includes a detector 63 and a detector 64. Here, fluorescence includes autofluorescence.

[0065] The first measurement sample prepared by the first sample preparation unit 5A and the second measurement sample prepared by the second sample preparation unit 5B, and the diluent contained in the reagent container 53 are supplied to the flow cell 61. In the flow cell 61, the first measurement sample and the second measurement sample are surrounded by the diluent as a sheath liquid.

[0066] The light source unit 62 irradiates the flow cell 61 with light having a wavelength band of 400 nm to 435 nm. Specifically, the light source unit 62 is a semiconductor laser light source, and irradiates the flow cell 61 with blue-violet laser light having a wavelength of 405 nm.

[0067] When light is irradiated on the sample flows of the first measurement sample and the second measurement sample in the flow chamber 61, the detectors 63 and 64 detect the light emitted from the first measurement sample and the second measurement sample, respectively. The respective sensitivity wavelength ranges of the detectors 63 and 64 are 400 nm to 1000 nm. The detectors 63 and 64 are avalanche photodiodes. In addition, other detectors such as laser diodes and photomultiplier tubes can also be used instead of avalanche photodiodes.

[0068] In the following description, the direction in which the light source unit 62 is connected to the flow cell 61 is referred to as the "X direction", and the direction in which the flow cell 61 is connected to the mirror 65 orthogonal to the X direction is referred to as the "Y direction". The detector 63 detects the fluorescence emitted from the first measurement sample and the second measurement sample respectively via the mirror 65 arranged on the Y direction side of the flow cell 61. In addition, the detector 64 is arranged on the X direction side of the flow cell 61, and is arranged on the opposite side of the light source unit 62 with the flow cell 61 interposed therebetween. The detector 64 detects the forward scattered light emitted from the measurement sample.

[0069] Furthermore, the device may be designed so that other scattered light such as side scattered light and back scattered light may be detected by a detector disposed at an appropriate position relative to the flow cell 61 instead of detecting the forward scattered light.

[0070] The detectors 63 and 64 each output an analog signal indicating the intensity of received light. Hereinafter, the analog signal output from the detector 63 is referred to as a "fluorescence signal", and the analog signal output from the detector 64 is referred to as a "forward scattered light signal".

[0071] The order in which the first and second measurement samples flow into the flow cell 61 is arbitrary. The first measurement sample may be first flowed into the flow cell 61, and then the second measurement sample may be flowed. Alternatively, the second measurement sample may be first flowed into the flow cell 61, and then the first measurement sample may be flowed.

[0072] The measurement section 2 further includes, for example, a cleaning device 8 that cleans the flow cell 61 after one of the first and second measurement samples passes through the flow cell 61 and before the other of the first and second measurement samples passes through the flow cell 61. The cleaning device 8 includes a metering pump such as a syringe pump and a diaphragm pump, and flows the diluent contained in the reagent container 53 to the flow cell 61 to clean the flow cell 61. When the first measurement sample is made to flow to the flow cell 61 first, the second measurement sample is made to flow to the flow cell 61 after the cleaning device 8 cleans the flow cell 61. When the second measurement sample is made to flow to the flow cell 61 first, the first measurement sample is made to flow to the flow cell 61 after the cleaning device 8 cleans the flow cell 61.

[0073] The measuring section 2 includes a second detecting device 7 for measuring red blood cells by a sheath flow DC detection method. Figure 2 As shown, the second detection device 7 is fluidically connected to the second reaction tank 54B via the electromagnetic valve 42. The second detection device 7 includes a sample nozzle 7a, an aperture member 7b, a recovery tube 7c and a detector 7d. A measurement sample (a third measurement sample) is supplied from the second reaction tank 54B to the sample nozzle 7a. The measurement sample supplied to the sample nozzle 7a passes through the aperture of the aperture member 7b, is recovered by the recovery tube 7c, and is sent to the waste liquid chamber 41. A voltage is applied to the measurement sample passing through the aperture of the aperture member 7b by the detector 7d. After the blood cells pass through the sheath flow pool, the voltage changes due to the resistance of the blood cells. The detector 7d captures the change in voltage and detects the resistance, thereby detecting the blood cells. The detector 7d outputs an analog signal representing the voltage. In addition, red blood cells can also be detected by scattered light detection in the optical detection device 6, so the second detection device 7 can also be omitted.

[0074] Back to Figure 1 The measuring unit 2 further includes a signal processing circuit 81 , a microcomputer 82 and a communication interface 83 .

[0075] The signal processing circuit 81 performs signal processing on the analog signals output by the detectors 63 and 64. The signal processing circuit 81 extracts the peak values ​​of the pulses contained in the fluorescence signal and the forward scattered light signal as characteristic parameters. Hereinafter, the peak value of the fluorescence signal is referred to as "fluorescence intensity", and the peak value of the forward scattered light signal is referred to as "forward scattered light intensity".

[0076] The microcomputer 82 controls the aspirating unit 4 , the first sample preparation unit 5A, the second sample preparation unit 5B, the optical detection device 6 , the signal processing circuit 81 , and the communication interface 83 .

[0077] The communication interface 83 is connected to the information processing unit 3 via a communication cable. The measurement unit 2 performs data communication with the information processing unit 3 via the communication interface 83. After the first measurement sample and the second measurement sample are measured, the communication interface 83 sends the first and second measurement data including characteristic parameters to the information processing unit 3.

[0078] The information processing unit 3 generates information related to malaria infection based on the intensity of fluorescence and scattered light detected from the first measurement sample, and generates information related to iron deficiency anemia based on the intensity of autofluorescence of erythrocytes detected from the second measurement sample.

[0079] Reference Figure 3 The structure of the information processing unit 3 is described. The information processing unit 3 includes a main body 300, an input device 309, and a display device 310. The main body 300 includes a CPU (Central Processing Unit) 301, a ROM (Read Only Memory) 302, a RAM (Random Access Memory) 303, a hard disk 304, a reader 305, an input / output interface 306, an image output interface 307, and a communication interface 308. The display device 310 displays text and images.

[0080] The CPU 301 executes the computer program stored in the ROM 302 and the computer program loaded in the RAM 303. The RAM 303 is used to read the computer program recorded in the ROM 302 and the hard disk 304. The RAM 303 is also used as a work area of ​​the CPU 301 when executing the computer program.

[0081] A computer program 320 for analyzing the first and second measurement data provided by the measurement unit 2 and outputting analysis results is installed in the hard disk 304 .

[0082] The reading device 305 is a floppy disk drive, a CD-ROM drive, a DVD-ROM drive, etc., and can read a computer program or data recorded in a removable recording medium 321. In addition, a computer program 320 for making the computer function as the information processing unit 3 is stored in the removable recording medium 321. The computer program 320 read from the removable recording medium 321 is installed in the hard disk 304.

[0083] The input device 309 includes input devices such as a keyboard and a mouse, and is connected to the input / output interface 306. The display device 310 is connected to the image output interface 307. The communication interface 308 is connected to the communication interface 83 of the measurement unit 2.

[0084] <Operation of blood analysis equipment>

[0085] Reference Figure 4 The operation of the blood analyzer 1 will be described.

[0086] First, the CPU 301 of the information processing unit 3 accepts an instruction to perform a measurement from the user via the input device 309 (step S101). After accepting the instruction to perform a measurement, the CPU 301 sends instruction data indicating the start of the measurement to the measurement unit 2 (step S102), and the measurement unit 2 receives the instruction data (step S103). The microcomputer 82 performs the first measurement sample preparation process (step S104), the first measurement process (step S105), the second measurement sample preparation process (step S106), and the second measurement process (step S107). In addition, the second measurement sample preparation process (step S106) and the second measurement process (step S107) may also be implemented before the first measurement sample preparation process (step S104) and the first measurement process (step S105).

[0087] Reference Figure 5 The first measurement sample preparation process (step S104) is described. The microcomputer 82 controls the aspiration unit 4 to supply a certain amount, for example, 17 μL of blood sample to the first reaction tank 54A (step S201). Next, the microcomputer 82 controls the first sample preparation unit 5A to supply a certain amount, for example, 1 mL of hemolytic agent from the reagent container 51 to the first reaction tank 54A, and to supply a certain amount, for example, 20 μL of nucleic acid staining reagent from the reagent container 52 to the first reaction tank 54A (step S202).

[0088] The first reaction tank 54A is heated to a certain temperature by the heater 55A. In the heated state, the mixture in the first reaction tank 54A is stirred by the pump 56A (step S203). After the blood sample is reacted with the hemolytic agent and the nucleic acid staining reagent, the microcomputer 82 controls the first sample preparation unit 5A to supply a certain amount of diluent from the reagent container 53 to the first reaction tank 54A to dilute the reaction solution of the blood sample and the hemolytic agent and the nucleic acid staining reagent, thereby preparing the first measurement sample. Next, the microcomputer 82 controls the first sample preparation unit 5A to export the first measurement sample from the first reaction tank 54A to the optical detection device 6 (step S204).

[0089] After the processing of step S204 is completed, the microcomputer 82 returns the processing to the main routine.

[0090] Refer to Figure 4In the first measurement process (step S105 ), the optical detection device 6 measures the first measurement sample. The first sample preparation unit 5A supplies the first measurement sample together with the sheath fluid to the flow cell 61 . The light source unit 62 irradiates the first measurement sample flow in the flow cell 61 with light.

[0091] When the first measurement sample flows in the flow chamber 61, blood cells including white blood cells, platelet aggregates and red blood cells pass through the flow chamber 61 in sequence. Here, "platelet aggregates" refer to aggregates of more than two platelets. When red blood cells infected with malaria (hereinafter referred to as "malaria-infected red blood cells") are contained, malarial parasites have entered the interior of the malarial-infected red blood cells. Due to the action of the hemolytic agent, the red blood cells in the measurement sample shrink. The malarial-infected red blood cells are shrunk by the hemolytic agent while having malarial parasites inside. In addition, malarial parasites are stained with nucleic acid staining reagents because they have nuclei. White blood cells are stained with staining reagents because they also have nuclei. Red blood cells that are not infected with malarial parasites (hereinafter referred to as "normal red blood cells") and platelet aggregates are hardly stained with nucleic acid staining reagents because they do not have nuclei.

[0092] Whenever light is irradiated on blood cells (leukocytes, platelet aggregates, and red blood cells), scattered light and fluorescence are emitted from the blood cells depending on the type of blood cells. The fluorescence emitted from the blood cells is detected by the detector 63. The scattered light (forward scattered light) emitted from the blood cells is detected by the detector 64.

[0093] The detectors 63 and 64 output electrical signals corresponding to the light reception levels as a fluorescence signal and a forward scattered light signal, respectively. The signal processing circuit 81 extracts the fluorescence intensity from the fluorescence signal and extracts the forward scattered light intensity from the forward scattered light signal. After the first measurement sample flows in the flow cell 61, the cleaning device 8 cleans the flow cell 61.

[0094] The second measurement sample preparation process (step S106) is described below. Figure 6 In step S301, the microcomputer 82 controls the aspirating unit 4 to aspirate a certain amount of blood sample from the test tube and supply 5 μL of the sample to the second reaction tank 54B. Next, in step S302, the microcomputer 82 controls the second sample preparation unit 5B to supply 1020 μL of the diluent from the reagent container 53 to the second reaction tank 54B.

[0095] The second reaction tank 54B has been heated to a certain temperature by the heater 55B. In the heated state, the mixture in the second reaction tank 54B is stirred by the pump 56B in step S303. Through the operations from steps S301 to S303, the second measurement sample is prepared in the second reaction tank 54B. In the second measurement sample, the blood cells are not hemolyzed and stained. In step S304, the microcomputer 82 controls the second sample preparation unit 5B to output the second measurement sample from the second reaction tank 54B to the optical detection device 6.

[0096] After the processing of step S304 is completed, the microcomputer 82 returns the processing to the main routine.

[0097] Refer to Figure 4 In the second measurement process (step S107 ), the optical detection device 6 measures the second measurement sample. The second measurement sample is supplied to the flow cell 61 together with the sheath fluid. The light source unit 62 irradiates the second measurement sample flow in the flow cell 61 with light.

[0098] When the second measurement sample flows in the flow chamber 61, blood cells including red blood cells pass through the flow chamber 61 in sequence. The amount of protoporphyrin in the red blood cells of healthy people is low, while the red blood cells of patients with iron deficiency anemia contain a lot of protoporphyrin. If the red blood cells containing a lot of protoporphyrin are irradiated with blue-violet laser, autofluorescence will be emitted. Autofluorescence is red light with a wavelength of more than 600nm and less than 700nm, so the autofluorescence emitted from each red blood cell is detected separately by the detector 63. On the other hand, even if the red blood cells that contain almost no protoporphyrin are irradiated with blue laser, almost no autofluorescence is generated. Therefore, the light receiving level of the detector 63 is a low value, and no autofluorescence is detected.

[0099] Whenever light is irradiated on red blood cells, scattered light is emitted from the red blood cells. The wavelength of the scattered light (forward scattered light) emitted from the red blood cells is 405 nm, and thus it is detected by the detector 64 .

[0100] The detector 63 and the detector 64 output electrical signals corresponding to the light reception levels as a fluorescence signal and a forward scattered light signal. The signal processing circuit 81 extracts the fluorescence intensity from the fluorescence signal and extracts the forward scattered light intensity from the forward scattered light signal. After the second measurement sample flows in the flow cell 61, the cleaning device 8 cleans the flow cell 61.

[0101] In the third measurement sample preparation process (step S108), the microcomputer 82 controls the aspirating unit 4 to aspirate a certain amount of blood sample from the test tube and supply 5 μL of the sample to the second reaction tank 54B. Next, the microcomputer 82 controls the second sample preparation unit 5B to supply 1020 μL of the diluent from the reagent container 53 to the second reaction tank 54B, thereby preparing the third measurement sample.

[0102] In the third measurement process (step S109), the second detection device 7 measures the third measurement sample. The second detection device 7 applies a voltage to the third measurement sample flowing in the sheath flow pool, and outputs an analog signal representing the voltage to the signal processing circuit 81. Whenever a red blood cell passes through the sheath flow pool, the voltage changes corresponding to its resistance. The signal processing circuit 81 detects the red blood cells by capturing the change in resistance through signal processing. Thus, the signal processing circuit 81 converts the output signal of the second detection device 7 into detection data of the red blood cells. After the third measurement process, the microcomputer 82 sends the first, second and third measurement data containing each characteristic parameter to the information processing unit 3 (step S110), and ends the process.

[0103] The information processing unit 3 receives the first, second and third measurement data (step S111). Next, the CPU 301 analyzes the red blood cell detection data included in the third measurement data and counts the red blood cells (step S112). After that, the CPU 301 performs the first measurement data analysis process for generating information related to malaria infection, generates the analysis result of the blood sample, and stores the analysis result in the hard disk 304 (step S113).

[0104] Reference Figure 7 The first measurement data analysis process is described below. When the first measurement data analysis process is started, first, the CPU 301 classifies the detected blood cells into white blood cells, platelet aggregates, normal red blood cells, and malaria-infected red blood cells based on the fluorescence intensity and forward scattered light intensity included in the first measurement data, and counts them separately (step S401).

[0105] Reference Figure 8 Explain the classification of white blood cells, platelet aggregation, normal red blood cells and malaria infected red blood cells. Figure 8 In the scatter plot, the vertical axis represents the forward scattered light intensity and the horizontal axis represents the fluorescence intensity.

[0106] White blood cells generally appear in region 401 where the forward scattered light intensity is greater than that of the group of normal red blood cells and malaria-infected red blood cells, and the fluorescence intensity is smaller than that of the group of normal red blood cells and platelet aggregation (the particle group belonging to region 401 is hereinafter referred to as the first particle group). Platelet aggregation generally appears in region 402 where the forward scattered light intensity is the same as that of region 401, and the fluorescence intensity is smaller than that of region 401. Normal red blood cells are greatly reduced in size by the hemolytic agent. Therefore, normal red blood cells reduced in size by the hemolytic agent generally appear in region 403 where the forward scattered light intensity is smaller than that of region 401, and the fluorescence intensity is smaller than that of region 401 (the particle group belonging to region 403 is hereinafter referred to as the second particle group).

[0107] Malaria-infected erythrocytes shrink due to the hemolytic agent, and because they contain malarial parasites with nuclei inside, they are stained by nucleic acid staining reagents. Therefore, malaria-infected erythrocytes generally appear in region 404 where the forward scattered light intensity is smaller than that of region 401 and the fluorescence intensity is greater than that of region 403 (the particle group belonging to region 404 is hereinafter referred to as the third particle group).

[0108] use Fig.10 and Fig.11 Describe the classification of subpopulations of malaria-infected erythrocytes. Fig.10 As shown, if a malaria mosquito with malaria parasites sucks blood, the protozoa will be injected into the blood along with the saliva of the malaria mosquito. The protozoa enter the liver cells, proliferate here, and are released into the blood again. The morphology of the protozoa at this time is called merozoite. After being released into the blood, it will immediately invade the red blood cells, and its morphology will continue to develop while changing. This morphological change is called life history, and each stage of the life history is called ring form, trophozoite, and schizont. Ring bodies include single ring bodies containing a single ring body in red blood cells, and multiple ring bodies containing several ring bodies in red blood cells. The protozoa that develop into schizonts destroy red blood cells, become merozoites again and are released into the blood. The released merozoites invade red blood cells, repeat the life history and proliferate again. Malaria parasites proliferate by repeating this cycle and continue to destroy red blood cells in the blood. In addition, some merozoites do not infect red blood cells and differentiate into a form called gametocytes. Gametocytes become the mother of further infection through the blood of malaria mosquitoes. The size and degree of staining of single ring bodies, multi-ring bodies, trophozoites, schizonts and gametocytes vary, so they can be Fig.11 The classification is performed in a scatter plot with the vertical axis being the forward scattered light intensity and the horizontal axis being the fluorescence intensity. Trophozoites or schizonts appear in the region 404 where malarial infected red blood cells appear, and the fluorescence intensity and forward scattered light intensity are higher than those of the ring bodies in region 413. Gametocytes appear in region 414 where the fluorescence intensity is lower than that of trophozoites or schizonts, and the forward scattered light intensity is the same as that of the trophozoites or schizonts with high forward scattered light intensity. Monocyclic bodies appear in region 411 where the fluorescence intensity is the same as that of the gametocytes with low fluorescence intensity, and the forward scattered light intensity is lower than that of the gametocytes. Polycyclic bodies appear in region 412 where the fluorescence intensity is higher than that of the monocyclic bodies, and the forward scattered light intensity is the same as that of the monocyclic bodies with low forward scattered light intensity.

[0109] Back to Figure 7 In step S402, CPU 301 executes malaria type determination processing.

[0110] Reference Fig. 9The malaria species determination process is described. The malaria species determination process is a process for determining whether a blood sample is suspected of being infected with Plasmodium falciparum or suspected of being infected with Plasmodium species other than Plasmodium falciparum (hereinafter referred to as "other species"). Plasmodium falciparum is a febrile disease with a high mortality rate, also known as tropical malaria, and distinguishing it from species other than Plasmodium falciparum has a high clinical significance. It is known that Plasmodium falciparum has more ring bodies than species other than Plasmodium falciparum.

[0111] In step S501, CPU 301 sets the malaria type·stage determination flag to 0. The malaria type·stage determination flag is set in a specific area of ​​RAM 303. The malaria type·stage determination flag set to 0 indicates that the blood sample is less likely to be infected with Plasmodium. The malaria type·stage determination flag set to 1 indicates that the blood sample is suspected of being infected with Plasmodium falciparum. The malaria type·stage determination flag set to 2 indicates that the blood sample is highly likely to be infected with Plasmodium falciparum. In addition, the suspicion of being infected with Plasmodium falciparum means that there is a possibility that Plasmodium falciparum has been infected, but the possibility cannot be said to be high. The malaria type·stage determination flag set to 3 indicates that the blood sample is suspected of being infected with other types of Plasmodium. The malaria type·stage determination flag set to 4 indicates that there is a high possibility that the blood sample is infected with other types of Plasmodium. In addition, the suspicion of being infected with other types of Plasmodium means that there is a possibility that Plasmodium falciparum has been infected, but the possibility cannot be said to be high.

[0112] In step S502, CPU301 determines whether the number of malaria-infected red blood cells counted in step S401 is above threshold value T1. Here, threshold value T1 is a value at which it can be determined that no malaria-infected red blood cells are present if the number of malaria-infected red blood cells is lower than T1. When the number of malaria-infected red blood cells is lower than T1 (no in step S502), it can be determined that the blood sample is not infected with Plasmodium. At this time, CPU301 ends the malaria type determination process and advances to the calculation process (step S403) of the malaria-infected red blood cell ratio.

[0113] After comparing the number of malaria-infected erythrocytes with the threshold value T1, if the number of malaria-infected erythrocytes is greater than T1 (YES in step S502), it is determined that the blood sample may have been infected with malarial parasites. At this time, CPU 301 transfers the process to step S503.

[0114] In step S503, CPU 301 Fig.11 The particle group belonging to area 411 in the scatter plot is counted as a single ring body, the particle group belonging to area 412 is counted as a polycyclic body, the particle group belonging to area 413 is counted as a trophozoite or a schizont (hereinafter referred to as "trophozoite·schizont"), and the particles belonging to area 414 are counted as gametocytes (step S503).

[0115] In step S504, CPU 301 determines whether the number of gametocytes is above threshold value T2. Threshold value T2 is a value that can be judged as no gametocytes if it is lower than T2. ​​When the number of gametocytes is above T2 (yes in step S504), it can be judged that gametocytes are present in the blood sample. At this time, CPU 301 sets the malaria type and stage determination flag to 2 (step S505), and transfers the process to step S506.

[0116] When the number of gametes is lower than T2 (No in step S504), it can be determined that gametes are not present in the blood sample. In this case, the CPU 301 transfers the process to step S506.

[0117] In step S506, CPU 301 determines whether the number of single ring bodies is above threshold value T3 (step S506). Threshold value T3 is a value that can be judged as no single ring body if it is lower than T3. When the number of single ring bodies is lower than T3 (no in step S506), CPU 301 transfers the process to step S511.

[0118] When the number of single ring bodies is equal to or greater than T3 (YES in step S506 ), CPU 301 determines whether the representative value of the forward scattered light intensity of region 411 where the single ring bodies appear is equal to or greater than threshold value T4 (step S507 ).

[0119] The size of the single ring of Plasmodium falciparum is smaller than that of the single ring of other types of Plasmodium. In addition, the forward scattered light intensity reflects the size of the cell. The representative value of the forward scattered light intensity of the region 411 where the single ring appears is the average value of the forward scattered light intensity of several particles appearing in the region 411. The threshold value T4 is a value that can be judged as being possibly infected with Plasmodium falciparum if it is lower than T4.

[0120] When the representative value of the forward scattered light intensity of region 411 is lower than T4 (yes in step S507), it is possible that Plasmodium falciparum is infected. At this time, CPU 301 determines whether the number of polycyclic bodies is above threshold value T5 (step S508). Threshold value T5 is a value that can be judged as no polycyclic body if the number of polycyclic bodies is lower than T5.

[0121] When the number of polycyclic bodies is less than T5 (No in step S508), it can be determined that polycyclic bodies do not appear in the blood sample. At this time, it cannot be said that the blood sample is likely to be infected with Plasmodium falciparum. On the other hand, in step S507, it is determined that the blood sample is likely to be infected with Plasmodium falciparum. In response, CPU 301 sets the malaria type and stage determination flag to 1 (step S509), ends the malaria type determination process, and returns to the first measurement data analysis process.

[0122] When the number of polycyclic bodies is greater than T5 (yes in step S508), it can be determined that polycyclic bodies may be present in the blood sample. At this time, it can be determined that the blood sample is likely to be infected with Plasmodium falciparum. In response, CPU 301 sets the malaria type and stage determination flag to 2 (step S510), ends the malaria type determination process, and returns to the first measurement data analysis process.

[0123] When the representative value of the forward scattered light intensity of the region 411 where the monocyclic body appears is above T4 (No in step S507), it is possible that the blood sample is infected with other types of Plasmodium. At this time, the CPU 301 determines whether the number of trophozoites and schizonts is above the threshold value T6 (step S511). The threshold value T6 is a value that can be judged as not appearing in the blood sample if it is lower than T6.

[0124] Trophozoites and schizonts are hardly found in the peripheral blood of patients infected with Plasmodium falciparum, while trophozoites and schizonts are found in the peripheral blood of patients infected with other types of Plasmodium. When the number of trophozoites and schizonts is lower than T6 (no in step S511), it can be determined that trophozoites and schizonts are not found in the blood sample. At this time, it cannot be said that the possibility that the blood sample is infected with other types of Plasmodium is high. On the other hand, in step S507, it is determined that the blood sample is suspected of being infected with other types of Plasmodium. In response to this, CPU 301 sets the malaria type and stage determination mark to 3 (step S512), terminates the malaria type determination process, and returns to the first measurement data analysis process.

[0125] When the number of trophozoites and schizonts is greater than T6 (Yes in step S511), it can be determined that trophozoites and schizonts are present in the blood sample. In addition, at this time, it can be determined that the blood sample is likely to be infected with other types of Plasmodium. In response, CPU 301 sets the malaria type and stage determination flag to 4 (step S513), ends the malaria type determination process, and returns to the first measurement data analysis process.

[0126] A scattergram of malaria type determination performed by the blood analyzer of this embodiment is taken as an example. Fig.12 The following is an example of a scatter plot of a group with a single ring and a group with multiple rings. Fig.12 As shown, the forward scattered light intensity and fluorescence intensity of the single ring group are smaller than those of the white blood cell group. In addition, the fluorescence intensity of the single ring group is smaller than that of the multi-ring group. Fig.13 is an example of a scatter plot of a population with monocyclic bodies and a population with gametophytes. Fig.13 As shown, the forward scattered light intensity of the gametocyte group is greater than that of the monocyclic group, and the fluorescence intensity is the same as or higher than that of the monocyclic group. Fig.14This is an example of a scatter plot of a group with single ring bodies and a group with trophozoites and schizonts. Fig.14 As shown, the fluorescence intensity of the trophozoite / schizont group is greater than that of the single ring body group.

[0127] Refer to Figure 7 After the above-mentioned malaria type determination process is completed, CPU 301 calculates the ratio of the number of malaria-infected red blood cells (the number of blood cells belonging to area 404) to the total number of red blood cells (the number of red blood cells counted in step S112) (hereinafter referred to as "malaria-infected red blood cell ratio" (step S403). Thus, CPU 301 ends the first measurement data analysis process and returns the process to Figure 4 The main program shown.

[0128] Next, CPU301 executes the second measurement data analysis process for generating information related to iron deficiency anemia, generates the analysis result of the blood sample, and stores the analysis result in hard disk 304 (step S114). In addition, CPU301 may also first execute the second measurement data analysis process (step S114), and then execute the first measurement data analysis process (step S113).

[0129] Reference Fig.15 The second measurement data analysis process is described. After the second measurement data analysis process is started, first, in step S601, the CPU 301 sets the iron deficiency anemia determination flag indicating the possibility of iron deficiency anemia to an initial value of 0. The iron deficiency anemia determination flag is stored in a specific area of ​​the RAM 303. When the iron deficiency anemia determination flag is set to 0, it indicates that the possibility of iron deficiency anemia is low, and when the iron deficiency anemia determination flag is set to 1, it indicates that the possibility of iron deficiency anemia exists.

[0130] In step S602, the CPU 301 extracts particles having a fluorescence intensity above a certain threshold value from the particle group determined to be red blood cells as red blood cells emitting autofluorescence (hereinafter referred to as "autofluorescent red blood cells"), and counts the autofluorescent red blood cells. The number of autofluorescent red blood cells is hereinafter referred to as "autofluorescent red blood cell count". In addition, red blood cells for which the fluorescence intensity above the threshold value is not detected are referred to as "red blood cells that do not emit autofluorescence".

[0131] use FIG. 16A to FIG. 16D The processing of step S602 is described below. Figure 7For the particle group belonging to region 403 in the scatter diagram generated in the processing of step S401 shown in FIG. 4, CPU 301 generates a scatter diagram 4000 with the forward scattered light intensity as the vertical axis and the fluorescence intensity as the horizontal axis. CPU 301 determines the particles belonging to region 400 where the forward scattered light intensity is higher than the noise region 490 and the fluorescence intensity is lower than the region 410 where the autofluorescent red blood cells appear as red blood cells that do not emit autofluorescence. CPU 301 determines the particles belonging to region 410 where the forward scattered light intensity is the same as that of region 400 and the fluorescence intensity is higher than that of region 400 as autofluorescent red blood cells. CPU 301 counts the particles belonging to region 410 to determine the number of autofluorescent red blood cells.

[0132] Fig.16A In the measurement result of a normal sample, that is, a blood sample collected from a healthy person, almost no particles appear in the region 410, and almost no particles regarded as autofluorescent red blood cells are detected. Fig. 16B This is the measurement result of a blood sample collected from a patient with iron deficiency anemia (hereinafter referred to as an "iron deficiency anemia sample"). Many particles appeared in the region 410, and many particles regarded as autofluorescent red blood cells were detected.

[0133] Fig. 16C and Fig.16D This is the measurement result of a blood sample collected from a thalassemia patient (hereinafter referred to as a "thalassemia sample"), and almost no particles appear in region 410, and almost no particles considered as autofluorescent red blood cells are detected. Thalassemia, which is classified as microcytic anemia like iron deficiency anemia, has similar symptoms and complete blood count (CBC) test values ​​to iron deficiency anemia, so it is clinically preferred to distinguish it from thalassemia and determine the possibility of iron deficiency anemia.

[0134] Back to Fig.15 In step S603, CPU 301 compares the number of autofluorescent red blood cells obtained in step S602 with the number of autofluorescent red blood cells obtained in step S112 ( Figure 4 ) is calculated as the autofluorescence information. FIG. 16A to FIG. 16D The total of the number of particles belonging to the region 400 and the number of particles belonging to the region 410 is shown as the number of red blood cells, instead of the number of red blood cells counted in step S112.

[0135] In step S604, the CPU 301 determines whether the autofluorescence ratio is greater than or equal to a threshold value T7. The threshold value T7 is a value that indicates the possibility of iron deficiency anemia if the autofluorescence ratio is greater than or equal to T7.

[0136] When the autofluorescence ratio is above T7 (Yes in step S604), in step S605, CPU 301 sets the iron deficiency anemia determination flag to 1, ends the second measurement data analysis process, and returns the process to the main program. When the autofluorescence ratio is lower than T7 (No in step S604), the iron deficiency anemia determination flag is still 0, ends the second measurement data analysis process, and returns the process to the main program.

[0137] Refer to Figure 4 After the above-mentioned second measurement data analysis process is completed, the CPU 301 causes the display device 310 to display the analysis result (step S115 ), and then ends the process.

[0138] Reference Fig.17 , the analysis result interface displayed by the display device 310 is described. The analysis result interface 500 includes a sample information display area 510, a patient information display area 520, a measurement result display area 530, and a reference information display area 540. The measurement result display area 530 includes a CBC item display area 531, a malaria item display area 532, and an autofluorescence item display area 533.

[0139] The sample information display area 510 displays blood sample information that is the origin of the analysis result displayed on the analysis result screen 500. The patient information display area 520 displays information of the subject from whom the blood sample has been collected.

[0140] The measurement result display area 530 displays the measurement values ​​of each item obtained by the measurement data analysis process. The CBC item display area 531 displays the measurement values ​​of basic measurement items in blood cell analysis. The measurement values ​​displayed in the CBC item display area 531 include the measurement values ​​of red blood cells (RBC) and white blood cells (WBC).

[0141] The malaria item display area 532 displays the measurement values ​​of the measurement items related to malaria-infected erythrocytes. The measurement values ​​displayed on the malaria item display area 532 include the measurement values ​​of the malaria-infected erythrocyte count (M-RBC) counted in step S401 and the malaria-infected erythrocyte ratio (MR) calculated in step S403. In addition, the malaria item display area 532 may display either the malaria-infected erythrocyte count (M-RBC) or the malaria-infected erythrocyte ratio (MR).

[0142] The autofluorescence item display area 533 displays the measurement values ​​of the measurement items related to the autofluorescence. The measurement values ​​displayed on the autofluorescence item display area 533 include the number of autofluorescent red blood cells (AF-RBC) counted in step S602 and the measurement value of the autofluorescence ratio (AF) calculated in step S603. In addition, either the number of autofluorescent red blood cells (AF-RBC) or the autofluorescence ratio (AF) can be displayed on the autofluorescence item display area 533.

[0143] The message displayed in the reference information display area 540 is determined by the combination of the malaria type and stage determination flag and the iron deficiency anemia determination flag. Fig.18 is a diagram showing the relationship between the combination of two marks and the displayed message. Fig.18 When the malaria type and stage determination flag and the iron deficiency anemia determination flag are 0, no message is displayed. When the malaria type and stage determination flag is 1 and the iron deficiency anemia determination flag is 0, the message is displayed. When the malaria type and stage determination mark is 2 and the iron deficiency anemia determination mark is 0, "P.falciparum+" is displayed as a message indicating that the possibility of falciparum malaria infection is high. When the malaria type and stage determination mark is 3 and the iron deficiency anemia determination mark is 0, "P.falciparum+" is displayed as a message indicating that the possibility of falciparum malaria infection is high. When the malaria type and stage determination mark is 4 and the iron deficiency anemia determination mark is 0, "O.Malaria+" is displayed as a message indicating that there is a high possibility of malaria infection other than falciparum malaria. When the malaria type and stage determination mark is 0 to 4 and the iron deficiency anemia determination mark is 1, "Iron deficiency anemia +" is displayed as a message indicating that there is a high possibility of malaria infection other than falciparum malaria. " as a message indicating that there is a suspicion of iron deficiency anemia. In addition, when the malaria type and stage judgment mark is 1 to 4 and the iron deficiency anemia judgment mark is 1, "Malaria&Iron alert" is displayed as a mark indicating that extra attention should be paid to providing iron supplements to the patient. This mark allows the inspector to easily make a judgment on whether to provide iron supplements to the patient.

[0144] According to the blood analyzer according to the present embodiment described above, information for determining whether to administer an iron supplement to an iron-deficiency anemia patient is generated by one blood analyzer, and thus measurement for determining whether to administer an iron supplement to an iron-deficiency anemia patient can be easily performed.

[0145] (Example)

[0146] Three C57BL / 6 mice (female, 6 weeks old; purchased from SLC Co., Ltd., Japan) were intraperitoneally injected with non-lethal red blood cells (3×105 After that, peripheral blood was collected from mice on days 4, 7, 10, 14, 17, 21, 28, 35, 42, and 49. After the collected peripheral blood was diluted 50 times with PBS, the ratio of the number of malaria-infected erythrocytes to the total number of erythrocytes, the number of autofluorescent erythrocytes, and the number of erythrocytes were measured using a multifunctional automatic blood cell analyzer XN-30 (manufactured by Sysmex Corporation). The results are as follows Fig.19 The multifunctional automatic blood cell analyzer XN-30 has the same structure as the optical detection device 6.

[0147] 14 days after infection, the ratio of malaria-infected red blood cells to the total number of red blood cells was more than 60%, and the number of autofluorescent red blood cells was 50×10 4 Red blood cells / μL, accounting for about 20% of the total red blood cell count. The scatter plots of day 0 and day 14 after infection are shown in Fig. 20 As shown, due to the infection, the number of red blood cells showing the stained fluorescence characteristics and scattered light characteristics characteristic of malaria infection increased, and the number of red blood cells showing the autofluorescence characteristics characteristic of iron deficiency anemia increased.

[0148] In addition, the fluorescence images of the blood of malaria-infected mice and non-infected mice were observed using a BZ-X710 fluorescence microscope (Keyence). As observation conditions, a BZ-X GFP filter (Ex = 470 / 40 nm, Em = 525 / 50 nm, dichroic mirror = 495 nm) was used for the purpose of detecting nucleic acids from malaria parasites, and a 5-ALA-405UF1-BLA filter (Ex = 405 / 20 nm, Em = 640 / 30 nm, dichroic mirror = 425 nm) was used for the purpose of detecting protoporphyrin IX-specific fluorescence.

[0149] The results are as follows Fig.21 As shown in the figure, no fluorescence from the dyeing of the nucleic acid of the malarial parasite and no autofluorescence from protoporphyrin IX were observed in the non-infected mice, while fluorescence from the dye staining the nucleic acid of the malarial parasite and autofluorescence from protoporphyrin IX were observed in the malarial infected mice. This result confirmed that the presence of malarial infected erythrocytes and erythrocytes containing protoporphyrin IX (iron-deficient erythrocytes) in malarial infected mice could be detected by the optical detection device 6 of the blood analysis device described in this embodiment.

[0150] Number Description

[0151] 1···blood analyzer, 2···measuring unit, 3···information processing unit, 5A, 5B···sample preparation unit, 6···optical detection device, 8···cleaning device, 7···second detection device, 61···flow chamber, 62···light source unit, 63, 64···detector, 310···display device.

Claims

1. A blood analysis device comprising: a first sample preparation unit that prepares a first measurement sample for measuring red blood cells infected with malaria from a blood sample; a second sample preparation unit that prepares a second measurement sample for measuring autofluorescence of red blood cells from the blood sample; a light source unit for irradiating light to the first measurement sample and the second measurement sample; a detection unit that detects fluorescence and scattered light generated by the first measurement sample irradiated with light, and detects autofluorescence generated by the second measurement sample irradiated with light; an information processing unit that generates information related to malaria infection based on the fluorescence and scattered light detected from the first measurement sample, and generates information related to iron deficiency anemia based on the autofluorescence detected from the second measurement sample; in, The information processing unit outputs display information for simultaneously outputting the malaria infection related information and the iron deficiency anemia related information to a display device, The information related to malaria infection includes the comparison result of the number of red blood cells infected with malaria or the ratio of the number of red blood cells infected with malaria to the number of red blood cells and the threshold value used to judge the possibility of malaria infection; the information related to iron deficiency anemia includes the comparison result of the number of red blood cells emitting autofluorescence or the ratio of the number of red blood cells emitting autofluorescence to the number of red blood cells and the threshold value used to judge the possibility of iron deficiency anemia; The information processing unit generates a mark for warning when the comparison result with the threshold value for judging the possibility of malaria infection indicates the possibility of malaria infection and the comparison result with the threshold value for judging the possibility of iron deficiency anemia indicates the possibility of iron deficiency anemia.

2. The blood analysis device according to claim 1, characterized in that: The first sample preparation unit prepares the first measurement sample by mixing the blood sample, a hemolytic agent that shrinks red blood cells, a dye that stains nucleic acid, and a diluent.

3. The blood analysis device according to claim 1 or 2, characterized in that: The second sample preparation unit prepares the second measurement sample by mixing the blood sample with a hemolytic agent that does not shrink erythrocytes and a dilution solution containing a dye that stains nucleic acid.

4. The blood analysis device according to claim 1 or 2, characterized in that: The blood analyzer further includes an aspirating unit that aspirates the blood sample from a sample container containing the blood sample; The first sample preparation section and the second sample preparation section prepare the first measurement sample and the second measurement sample, respectively, from a portion of the blood sample aspirated by the aspirating section.

5. The blood analysis device according to claim 1 or 2, characterized in that: The wavelength band of the light emitted by the light source is not less than 400 nm and not more than 435 nm.

6. The blood analysis device according to claim 1 or 2, characterized in that: The blood analyzer further comprises a flow chamber for allowing the first and second measurement samples to flow; The light source unit irradiates light to the first and second measurement samples flowing in the flow cell; The detection unit detects fluorescence and scattered light generated by the first measurement sample flowing through the flow cell, and detects autofluorescence generated by the second measurement sample flowing through the flow cell.

7. The blood analysis device according to claim 6, characterized in that: The blood analyzer further includes a cleaning device configured to clean the flow cell after one of the first and second measurement samples passes through the flow cell and before the other of the first and second measurement samples passes through the flow cell.

8. The blood analysis device according to claim 1 or 2, characterized in that: The malaria infection-related information includes the number of red blood cells infected with malaria, or the ratio of the number of red blood cells infected with malaria to the number of red blood cells.

9. The blood analysis device according to claim 1 or 2, characterized in that: The malaria infection-related information includes information indicating the possibility of malaria infection based on a comparison result of the number of malaria-infected red blood cells or the ratio of the number of malaria-infected red blood cells to the number of red blood cells and a threshold value for determining the possibility of malaria infection.

10. The blood analysis device according to claim 9, characterized in that: The information indicating the possibility of malaria infection includes information indicating the type of malaria infected.

11. The blood analysis device according to claim 1 or 2, characterized in that: The information related to iron deficiency anemia includes the number of red blood cells emitting autofluorescence, or the ratio of the number of red blood cells emitting autofluorescence to the number of red blood cells.

12. The blood analysis device according to claim 1 or 2, characterized in that: The information related to iron deficiency anemia includes information indicating the possibility of iron deficiency anemia based on a comparison result of the number of red blood cells emitting autofluorescence or the ratio of the number of red blood cells emitting autofluorescence to the number of red blood cells and a threshold value for determining the possibility of iron deficiency anemia.

Images