Sample analyzer and sample analysis method

Abstract

The embodiments of the present application relate to a sample analyzer and a sample analysis method. The sample analysis method comprises: aspirating a blood sample to be tested; mixing at least a portion of said blood sample, a hemolytic agent, a first fluorescent dye and a second fluorescent dye to prepare a measurement sample; in a single test, making each particle in the measurement sample pass through an optical detection zone irradiated by excitation light, so as to acquire optical information generated after each particle in the measurement sample has been irradiated by the excitation light, wherein the excitation light comprises light of a first wavelength capable of exciting the first fluorescent dye and light of a second wavelength capable of exciting the second fluorescent dye, and the second wavelength is greater than the first wavelength; on the basis of at least one scattered light signal and a first fluorescence signal, obtaining infected red blood cell information of the measurement sample; and on the basis of the at least one scattered light signal and a second fluorescence signal, obtaining a four-part white blood cell differential result of the measurement sample. Therefore, infected red blood cell information and a white blood cell differential result of a measurement sample can be obtained at the same time in a single test.

Description

Sample analyzer and sample analysis method TECHNICAL FIELD

[0001] The present application relates to the field of blood cell detection, and in particular to a sample analyzer and a sample analysis method. BACKGROUND

[0002] Malaria is a serious parasitic disease that seriously endangers human health caused by Plasmodium parasitizing in human body, which is mainly transmitted in humans and anopheles by female anopheles biting human body infected with Plasmodium, and is one of the most serious tropical insect-borne infectious diseases in the world so far, with a high mortality rate. If malaria is not timely differentiated from other febrile diseases for treatment, malaria can develop into a serious disease within 24 hours, causing illness delay, leading to severe and death cases.

[0003] In the related art, blood cell counting using a blood cell analyzer is a routine detection item for patients with fever.

[0004] With the development of blood cell analysis technology in recent years, blood cell analyzers have developed from single-cell counting detection to multi-functional joint detection instruments that can simultaneously analyze tens of thousands of cells at high speed and obtain multiple parameters describing cell morphological characteristics. Using digital and image results in blood analyzers helps to screen malaria, but the application of blood cell analyzers in the related art in malaria diagnosis still has many limitations. SUMMARY

[0005] One task of the present application is to obtain infected red blood cell information and white blood cell classification results through only one reaction and detection channel.

[0006] Another task of the present application is to obtain infected red blood cell information and nucleated red blood cell information through only one reaction and detection channel.

[0007] To achieve the tasks of the present application, the first aspect of the present application relates to a sample analyzer, comprising:

[0008] a sample suction device for sucking a blood sample to be tested;

[0009] a sample preparation device for mixing at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye and a second fluorescent dye in a reaction pool to prepare a test sample;

[0010] An optical detection device comprises a light emitting device for emitting excitation light to irradiate a flow cell, a flow chamber in communication with the flow cell and for passing each particle in the assay sample therethrough, and a light detector for detecting optical information generated by each particle in the assay sample when irradiated by the excitation light in a single test, the light emitting device comprising a first light source for emitting light of a first wavelength capable of exciting the first fluorescent dye and a second light source for emitting light of a second wavelength capable of exciting the second fluorescent dye, wherein the second wavelength is greater than the first wavelength, the optical information comprising at least one scatter light signal, a first fluorescent signal corresponding to the first fluorescent dye, and a second fluorescent signal corresponding to the second fluorescent dye detected in the single test; and

[0011] a processor configured to obtain infected red blood cell information of the assay sample based on at least one of the scatter light signals and the first fluorescent signal, and obtain a white blood cell four-classification result of the assay sample based on at least one of the scatter light signals and the second fluorescent signal, wherein the white blood cell four-classification result comprises a count result of a lymphocyte population, a count result of a monocyte population, a count result of a neutrophil population, and a count result of an eosinophil population in the assay sample.

[0012] To achieve the object of the present application, the second aspect of the present application relates to a sample analysis method, comprising:

[0013] aspirating a blood sample to be tested;

[0014] mixing at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye, and a second fluorescent dye to prepare an assay sample;

[0015] passing each particle in the assay sample through an optical detection region irradiated by excitation light in a single test to obtain optical information generated by each particle in the assay sample when irradiated by the excitation light, the excitation light comprising light of a first wavelength capable of exciting the first fluorescent dye and light of a second wavelength capable of exciting the second fluorescent dye, wherein the second wavelength is greater than the first wavelength, the optical information comprising at least one scatter light signal, a first fluorescent signal corresponding to the first fluorescent dye, and a second fluorescent signal corresponding to the second fluorescent dye obtained in the single test;

[0016] obtaining infected red blood cell information of the assay sample based on at least one of the scatter light signals and the first fluorescent signal, and

[0017] obtaining the white blood cell four-classification result of the test sample based on at least one of the scattering light signals and the second fluorescence signal, wherein the white blood cell four-classification result comprises a count result of a lymphocyte group, a count result of a monocyte group, a count result of a neutrophil group, and a count result of an eosinophil group in the test sample.

[0018] In the technical solutions provided in the first aspect and the second aspect of the present application, the blood sample to be tested, the hemolytic agent, and the two fluorescent dyes are mixed to prepare a test sample in one reaction and detection channel, and the test sample is subjected to optical determination in one test, so that the information of infected red blood cells and the white blood cell four-classification result are obtained based on the optical information obtained in one test.

[0019] To achieve the object of the present application, the third aspect of the present application relates to a sample analyzer, comprising:

[0020] a sample suction device configured to suck the blood sample to be tested;

[0021] a sample preparation device configured to mix at least part of the blood sample to be tested, the hemolytic agent, the first fluorescent dye, the second fluorescent dye, and the third fluorescent dye in a reaction cell to prepare a test sample;

[0022] an optical detection device comprising a light emitting device, a flow chamber, and a light detector, the light emitting device being configured to emit excitation light to irradiate the flow chamber, the flow chamber being in communication with the reaction cell and being configured to allow each particle in the test sample to pass therethrough, the light detector being configured to detect optical information generated by each particle in the test sample when the particle passes through the flow chamber and is irradiated by the excitation light in one test, the light emitting device comprising a first light source and a second light source, the first light source being configured to emit light of a first wavelength capable of exciting the first fluorescent dye, the second light source being configured to emit light of a second wavelength capable of exciting the third fluorescent dye, the light of the first wavelength or the light of the second wavelength also being capable of exciting the second fluorescent dye, wherein the second wavelength is greater than the first wavelength, and the optical information comprises at least one scattering light signal, a first fluorescence signal corresponding to the first fluorescent dye, a second fluorescence signal corresponding to the second fluorescent dye, and a third fluorescence signal corresponding to the third fluorescent dye detected in the one test; and

[0023] The processor is configured to obtain the information of infected red blood cells and the white blood cell five-classification result of the test sample based on at least one of the scattering light signals, the first fluorescence signal, the second fluorescence signal and the third fluorescence signal, wherein the white blood cell five-classification result includes the count result of the lymphocyte group, the count result of the monocyte group, the count result of the neutrophil group, the count result of the eosinophil group and the count result of the basophil group in the test sample.

[0024] To achieve the object of the present application, the fourth aspect of the present application relates to a sample analysis method, comprising:

[0025] Aspirating a blood sample to be tested;

[0026] Mixing at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye, a second fluorescent dye and a third fluorescent dye to prepare a test sample;

[0027] Passing each particle in the test sample through an optical detection area irradiated by excitation light in a single test to obtain optical information generated by each particle in the test sample after being irradiated by the excitation light, the excitation light including light of a first wavelength capable of exciting the first fluorescent dye and light of a second wavelength capable of exciting the third fluorescent dye, the light of the first wavelength or the light of the second wavelength also being capable of exciting the second fluorescent dye, wherein the second wavelength is greater than the first wavelength, and the optical information includes at least one scattering light signal, a first fluorescence signal corresponding to the first fluorescent dye, a second fluorescence signal corresponding to the second fluorescent dye and a third fluorescence signal corresponding to the third fluorescent dye obtained in the single test; and

[0028] Obtaining the information of infected red blood cells and the white blood cell five-classification result of the test sample based on at least one of the scattering light signals, the first fluorescence signal, the second fluorescence signal and the third fluorescence signal, wherein the white blood cell five-classification result includes the count result of the lymphocyte group, the count result of the monocyte group, the count result of the neutrophil group, the count result of the eosinophil group and the count result of the basophil group in the test sample.

[0029] In the technical solutions provided in the third aspect and the fourth aspect of the present application, the blood sample to be tested, the hemolytic agent and the three fluorescent dyes are mixed to prepare a test sample in one reaction and detection channel, and the test sample is optically determined in one test, so as to obtain the information of infected red blood cells and the white blood cell five-classification result according to the optical information obtained in one test.

[0030] To achieve the object of the present application, the fifth aspect of the present application relates to a sample analyzer, comprising:

[0031] a sample preparation device for mixing at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye and a third fluorescent dye in a reaction cell to prepare an assay sample;

[0032] a sample preparation device for mixing at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye and a third fluorescent dye in a reaction cell to prepare an assay sample;

[0033] an optical detection device comprising a light emitting device for emitting excitation light to irradiate a flow cell, the flow cell being in communication with the reaction cell and for passing each particle in the assay sample therethrough, and a light detector for detecting optical information generated by each particle in the assay sample when irradiated by the excitation light in a single test, the optical information comprising at least one scattering light signal, a first fluorescent signal corresponding to the first fluorescent dye and a third fluorescent signal corresponding to the third fluorescent dye detected in the single test; and

[0034] a processor configured to obtain infected red blood cell information and nucleated red blood cell information of the assay sample based on at least one of the scattering light signal, the first fluorescent signal and the third fluorescent signal.

[0035] To achieve the object of the present application, the sixth aspect of the present application relates to a sample analysis method, comprising:

[0036] a blood sample to be tested is aspirated;

[0037] at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye and a third fluorescent dye are mixed to prepare an assay sample;

[0038] each particle in the assay sample is passed through an optical detection region irradiated by excitation light in a single test to obtain optical information generated by each particle in the assay sample when irradiated by the excitation light, the optical information comprising at least one scattering light signal, a first fluorescent signal corresponding to the first fluorescent dye and a third fluorescent signal corresponding to the third fluorescent dye obtained in the single test; and

[0039] infected red blood cell information and nucleated red blood cell information of the assay sample are obtained based on at least one of the scattering light signal, the first fluorescent signal and the third fluorescent signal.

[0040] In the technical solutions provided in the fifth aspect and the sixth aspect of the present application, in one reaction and detection channel, a blood sample to be tested, a hemolytic agent and two dyes are mixed to prepare an assay sample, and optical determination is performed on the assay sample in one test, so as to obtain infected red blood cell information and nucleated red blood cell information according to optical information obtained in one test. BRIEF DESCRIPTION OF DRAWINGS

[0041] Fig. 1 is a structural schematic diagram of one embodiment of a sample analyzer according to the present application.

[0042] Fig. 2 is a structural schematic diagram of one embodiment of an optical detection device according to the present application.

[0043] Fig. 3 is a schematic flow chart of one embodiment of a sample analysis method according to the present application.

[0044] Fig. 4 is a schematic flow chart of another embodiment of a sample analysis method according to the present application.

[0045] Fig. 5 is a schematic flow chart of yet another embodiment of a sample analysis method according to the present application.

[0046] Figs. 6a, 6b and 6c are scatter plots obtained by detecting a blood sample to be tested which is not infected with Plasmodium according to the first embodiment of the present application.

[0047] Figs. 7a, 7b and 7c are scatter plots obtained by detecting a blood sample to be tested which is infected with Plasmodium according to the first embodiment of the present application.

[0048] Figs. 8a, 8b and 8c are scatter plots obtained by detecting a blood sample to be tested according to the second embodiment of the present application.

[0049] Figs. 9a, 9b and 9c are scatter plots obtained by detecting a blood sample to be tested according to the third embodiment of the present application.

[0050] Figs. 10a, 10b, 10c and 10d are scatter plots obtained by detecting a blood sample to be tested according to the fourth embodiment of the present application.

[0051] Figs. 11a and 11b are scatter plots obtained by detecting a blood sample to be tested according to the fifth embodiment of the present application. DETAILED DESCRIPTION

[0052] The embodiments of the present application will be described below in detail with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of the present application, but not all the embodiments. Based on the embodiments in the present application, all the other embodiments obtained by those skilled in the art without creative work fall within the scope of the present application.

[0053] In the present application, the serial numbers of components, such as "first", "second", etc., are used only to distinguish the described objects, and do not have any sequence or technical meaning. The "connection" and "coupling" in the present application include direct and indirect connection (coupling) unless otherwise specified.

[0054] For the convenience of the following description, some terms involved in the present application are first explained briefly as follows:

[0055] 1) Scattergram: a two-dimensional or three-dimensional graph generated by a blood cell analyzer, on which a plurality of two-dimensional or three-dimensional characteristic information of particles are distributed. The X-axis, Y-axis and Z-axis of the scattergram each represent a characteristic of each particle. For example, in a scattergram, the X-axis represents the intensity of forward scattering light, the Y-axis represents the intensity of fluorescence, and the Z-axis represents the intensity of side scattering light.

[0056] The term "scattergram" used in the present application not only refers to a distribution graph of at least two groups of data in the form of data points in a rectangular coordinate system, but also includes a data array, i.e. is not limited by the form of its graph presentation.

[0057] 2) Cell population: a particle population formed by a plurality of particles with the same characteristics distributed in a certain area of a scattergram, also known as "particle group" or "cell group", such as a white blood cell (including all types of white blood cells) group, and a white blood cell subpopulation, such as a neutrophil group, a lymphocyte group, a monocyte group, an eosinophil group, or a basophil group.

[0058] 3) Blood shadow: a fragment particle obtained by lysing red blood cells and platelets in blood with a hemolytic agent.

[0059] The blood cell analyzer used in the present application classifies and counts particles in a blood sample by combining laser scattering method and fluorescence staining method of flow cytometry. Here, the principle of detecting the blood sample by the blood cell analyzer can be, for example, as follows: first, a blood sample is taken and treated with a hemolytic agent and a fluorescent dye, wherein red blood cells are destroyed and lysed by the hemolytic agent, while white blood cells are not lysed, but the fluorescent dye can enter the cell nucleus of the white blood cells with the help of the hemolytic agent and bind to the nucleic acid substances in the cell nucleus; then the particles in the sample pass through the detection hole irradiated by a laser beam one by one, when the laser beam irradiates the particles, the characteristics of the particles (such as volume, staining degree, cell content size and content, nuclear density, etc.) can block or change the direction of the laser beam, thereby generating scattering light of various angles corresponding to the characteristics of the particles. After the scattering light is received by a signal detector, information related to the structure and composition of the particles can be obtained. Among them, the forward scattering light (Forward scatter, FS) reflects the number and volume of the particles, the side scattering light (Side scatter, SS) reflects the complexity of the internal structure of the cells (such as intracellular particles or cell nucleus), and the fluorescence (Fluorescence, FL) reflects the content of nucleic acid substances in the cells. Using these light information, the cells in the blood sample can be classified and counted.

[0060] FIG. 1 is a schematic diagram of a sample analyzer according to some embodiments of the present application. The sample analyzer 100 includes at least a sample suction device 110, a sample preparation device 120, an optical detection device 130, and a processor 140. The sample suction device 110 is configured to suction a blood sample to be tested. The sample preparation device 120 is configured to mix at least a portion of the blood sample to be tested, a hemolytic agent, and at least two fluorescent dyes in a reaction cell to prepare an assay sample. The optical detection device 130 includes a light emitting device, a flow cell, and a light detector. The light emitting device is configured to emit excitation light to illuminate the flow cell, the flow cell is in communication with the reaction cell and is configured to allow each particle in the assay sample to pass through, and the light detector is configured to detect optical information generated by each particle in the assay sample when illuminated by the excitation light in a single test. The optical information includes at least one scattering light signal and at least two fluorescent signals detected in a single test.

[0061] In some embodiments, the sample suction device 110 has a sampling needle (not shown) configured to suction the blood sample to be tested. In addition, the sample suction device 110 can further include a driving device configured to drive the sampling needle to quantitatively suction the blood sample to be tested from a test tube through a needle nozzle of the sampling needle. In addition, the sample suction device 110 can further deliver the suctioned blood sample to be tested to the sample preparation device 120.

[0062] In some embodiments, the sample preparation device 120 can include at least one reaction cell and a reagent supply device (not shown). The at least one reaction cell is configured to receive the blood sample to be tested suctioned by the sample suction device 110, and the reagent supply device is configured to provide processing reagents (including a hemolytic agent, fluorescent dyes, etc.) to the at least one reaction cell, so that the blood sample to be tested suctioned by the sample suction device 110 is mixed with the processing reagents provided by the reagent supply device in the reaction cell to prepare an assay sample.

[0063] For example, the sample preparation device 120 is configured to mix and incubate the blood sample, the hemolytic agent, and at least two fluorescent dyes, with a reaction time of 10s-1min and a reaction temperature of 25℃-50℃.

[0064] In some embodiments, the sample preparation device 120 is configured to mix at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye, and a second fluorescent dye in a reaction cell to prepare an assay sample.

[0065] In some other embodiments, the sample preparation device 120 is configured to mix at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye, and a third fluorescent dye in a reaction cell to prepare an assay sample.

[0066] In yet other embodiments, the sample preparation device 120 is configured to mix at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye, a second fluorescent dye, and a third fluorescent dye in a reaction cell to prepare an assay sample.

[0067] In embodiments of the present application, the hemolytic agent can dissolve red blood cells and platelets in the blood sample, but can keep the morphology of white blood cells substantially unchanged. The fragments of red blood cells and platelets obtained after being dissolved form a blood shadow, thereby distinguishing from white blood cells and nucleated red blood cells in the blood sample.

[0068] In some embodiments, the hemolytic agent can include any one or a combination of a cationic surfactant, a non-ionic surfactant, an anionic surfactant, an amphiphilic surfactant, a buffer pair. The cationic surfactant is for example selected from at least one or a combination of dodecyltrimethylammonium chloride, octyltrimethylammonium bromide, tetradecyltrimethylammonium chloride. The non-ionic surfactant is for example selected from at least one or a combination of long-chain fatty alcohol polyoxyethylene, alkylphenol polyoxyethylene ether, fatty acid polyoxyethylene ether, fatty amine polyoxyethylene ether. The buffer pair is for example selected from at least one or a combination of phosphate, citrate, Tris-HCl, MES buffer. The anionic surfactant is for example selected from at least one or a combination of dodecylbenzenesulfonic acid, sodium fatty alcohol acylsulfate, sodium ethoxylated fatty acid methyl ester sulfonate, sodium secondary alkyl sulfonate, alcohol ether carboxylate.

[0069] In some embodiments, the above-mentioned hemolytic agent can include a buffer capable of adjusting the pH value of the hemolytic agent to between 5-11, preferably to between 6-8.

[0070] In other embodiments, the hemolytic agent can include at least one of alkyl glycoside, triterpenoid saponin, steroidal saponin.

[0071] In embodiments of the present application, the hemolytic agent can allow blood cells to have good permeability under short hemolysis time processing, and then the fluorescent dye enters the cells and specifically binds and stains the cells. Due to the difference in the ability of nucleic acids in white blood cells and nucleated red blood cells and cytochrome, riboflavin, etc. inside red blood cells and platelets to bind with the fluorescent dye, different fluorescent signals will be generated.

[0072] In some embodiments, the first fluorescent dye can be a fluorescent dye capable of specifically labeling Plasmodium, which includes a compound having a structural formula as follows:

[0073] In the structural formula,

[0074] R1 is selected from substituted or unsubstituted phenyl;

[0075] R2is selected from C 1-8 a nitrogen-containing or oxygen-containing heterocycle, which is substituted with a substituted or unsubstituted phenyl;

[0076] X is halogen or PF6-;

[0077] each of the phenyl groups of R1and R2is independently optionally substituted with a substituent selected from CN, COOH, NH2, NO2, OH, SH, C 1-6 alkoxy, C 1-6 alkylamino, C 1-6 amide, halogen or C 1-6 haloalkyl.

[0078] Further, R1is selected from hydrogen, phenyl or biphenyl. Still further, R1is selected from biphenyl.

[0079] Further, R2is selected from N, N-dimethyl, piperazinyl or 2,6-dimethylpiperazinyl. Still further, R2is selected from 2,6-dimethylpiperazinyl.

[0080] As some embodiments, the compound for the first fluorescent dye has one of the following structures A, B, C, D:

[0081] Compound A

[0082] Compound B

[0083] Compound C

[0084] Compound D

[0085] Here, the concentration of the first fluorescent dye is 0.1-1000 mg / L.

[0086] For more embodiments of the first fluorescent dye of the present application, reference can be made to the Applicant’s Chinese application CN202310028120.1 filed on January 9, 2023, the entire disclosure of which is incorporated herein by reference.

[0087] In some embodiments, the second fluorescent dye can be a fluorescent dye that can be used for white blood cell differential, which includes a compound having the following general structure:

[0088] Here, the concentration of the second fluorescent dye is 0.1-1000 mg / L.

[0089] For more embodiments of the second fluorescent dye of the present application, reference can be made to the Applicant’s US patent US7598385B2, the entire disclosure of which is incorporated herein by reference.

[0090] In some embodiments, the third fluorescent dye can be a fluorescent dye capable of identifying nucleated red blood cells, which includes a compound having a structural formula as follows:

[0091] In the structural formula, X is selected from one of -O-, -S- and ; R1and R2are each independently selected from C3-C 20 alkenyl and C3-C 20 alkyne.

[0092] As some implementations, the compound for the third fluorescent dye has one of the following structures I, II, III, IV:

[0093] Compound I

[0094] Compound II

[0095] Compound III

[0096] Compound IV

[0097] Compound V

[0098] Here, the concentration of the third fluorescent dye is 0.1-1000 mg / L.

[0099] In some embodiments, the first fluorescent dye and / or the second fluorescent dye and / or the third fluorescent dye is preserved in a water-soluble organic phase such as glycerol, glycol, ethylene glycol, etc.

[0100] In some embodiments, the first fluorescent dye and / or the second fluorescent dye and / or the third fluorescent dye can be preserved alone or mixed with a hemolytic agent.

[0101] In some embodiments, the light emitting device includes a first light source for emitting light of a first wavelength capable of exciting the first fluorescent dye and a second light source for emitting light of a second wavelength capable of exciting the second fluorescent dye, wherein the second wavelength is greater than the first wavelength. The optical information includes at least one of a scattering light signal detected in a single test, a first fluorescent signal corresponding to the first fluorescent dye and a second fluorescent signal corresponding to the second fluorescent dye.

[0102] In other embodiments, the light emitting device comprises a first light source for emitting light of a first wavelength capable of exciting a first fluorescent dye and a second light source for emitting light of a second wavelength capable of exciting a third fluorescent dye, wherein the second wavelength is greater than the first wavelength. The optical information comprises at least one scattered light signal detected in a single test, a first fluorescent signal corresponding to the first fluorescent dye and a third fluorescent signal corresponding to the third fluorescent dye.

[0103] In yet other embodiments, the light emitting device comprises a first light source for emitting light of a first wavelength capable of exciting a first fluorescent dye and a second dye and a second light source for emitting light of a second wavelength capable of exciting a third fluorescent dye, wherein the second wavelength is greater than the first wavelength. The optical information comprises at least one scattered light signal detected in a single test, a first fluorescent signal corresponding to the first fluorescent dye, a second fluorescent signal corresponding to the second fluorescent dye and a third fluorescent signal corresponding to the third fluorescent dye.

[0104] In yet other embodiments, the light emitting device comprises a first light source for emitting light of a first wavelength capable of exciting a first fluorescent dye and a second dye and a second light source for emitting light of a second wavelength capable of exciting a third fluorescent dye, wherein the second wavelength is greater than the first wavelength. The optical information comprises at least one scattered light signal detected in a single test, a first fluorescent signal corresponding to the first fluorescent dye, a second fluorescent signal corresponding to the second fluorescent dye and a third fluorescent signal corresponding to the third fluorescent dye.

[0105] In some embodiments, the flow chamber in the optical detection device 130 refers to a chamber adapted to detect a focused fluid flow of scattered light signals and fluorescent signals. When a particle, such as a blood cell, passes through a detection aperture of the flow chamber, the particle scatters the incident light beam directed to the detection aperture from the light source in various directions. Light detectors can be disposed at one or more different angles relative to the incident light beam to detect the light scattered by the particle, thereby obtaining a scattered light pulse signal (also referred to as a scattered light signal). Since different particles have different light scattering characteristics, the scattered light signal can be used to distinguish different populations of particles.

[0106] In particular, the detected scattered light signal in the vicinity of the incident light beam is often referred to as a forward scattered light signal or a small angle scattered light signal. In some embodiments, the forward scattered light signal can be detected at an angle of about 1° to about 10° from the incident light beam. In other embodiments, the forward scattered light signal can be detected at an angle of about 2° to about 6° from the incident light beam. The detected scattered light signal in a direction about 90° from the incident light beam is often referred to as a side scattered light signal. In some embodiments, the side scattered light signal can be detected at an angle of about 65° to about 115° from the incident light beam. Typically, the fluorescent signal emitted from a blood cell stained with a fluorescent dye is also detected in a direction about 90° from the incident light beam.

[0107] In some embodiments, the light detectors in the optical detection device 130 can comprise a scattered light detector, e.g. a side scattered light detector, for detecting a scattered light pulse signal, e.g. a side scattered light pulse signal, and a first fluorescent detector for detecting a first fluorescent pulse signal, e.g. a first fluorescent signal, and a second fluorescent detector for detecting a second fluorescent pulse signal, e.g. a second fluorescent signal.

[0108] In other embodiments, the light detectors in the optical detection device 130 can comprise a scattered light detector, e.g. a side scattered light detector, for detecting a scattered light pulse signal, e.g. a side scattered light pulse signal, and a first fluorescent detector for detecting a first fluorescent pulse signal, e.g. a first fluorescent signal, and a third fluorescent detector for detecting a third fluorescent pulse signal, e.g. a third fluorescent signal.

[0109] In yet other embodiments, the light detectors in the optical detection device 130 can comprise a scattered light detector, e.g. a side scattered light detector, for detecting a scattered light pulse signal, e.g. a side scattered light pulse signal, and a first fluorescent detector for detecting a first fluorescent pulse signal, e.g. a first fluorescent signal, a second fluorescent detector for detecting a second fluorescent pulse signal, e.g. a second fluorescent signal, and a third fluorescent detector for detecting a third fluorescent pulse signal, e.g. a third fluorescent signal.

[0110] In some embodiments, the optical detection device 130 comprises a forward scattered light detector for detecting a forward scattered light signal or a side scattered light detector for detecting a side scattered light signal. Preferably, the optical detection device 130 comprises both a forward scattered light detector and a side scattered light detector.

[0111] Figure 2 is a specific example of the optical detection device 130. The optical detection device 130 has a light emitting device 131, a forward light assembly 132, a flow chamber 133, a forward scattering light detector 134, a first dichroic mirror 135, a side scattering light detector 136, a second dichroic mirror 137, a first fluorescence detector 138, and a second fluorescence detector 139. The first fluorescence detector 138 is configured to detect a first fluorescence signal corresponding to a first fluorescence dye from a particle passing through the flow chamber 133 after being irradiated by a light beam, and the second fluorescence detector 139 is configured to detect a second fluorescence signal corresponding to a second fluorescence dye from a particle passing through the flow chamber 133 after being irradiated by a light beam. Here, the light emitting device 131, the forward light assembly 132, the flow chamber 133, and the forward scattering light detector 134 are arranged in sequence along an optical axis. The forward light assembly 132 is configured to focus a light beam emitted by the light emitting device 131 on a detection region of the flow chamber 133 in a direction of particle flow, so that a particle flowing through the detection region of the flow chamber 133 can generate scattered light. On one side of the flow chamber 133, the first dichroic mirror 135 is arranged at an angle of 45° to the optical axis. A portion of the side light generated by the particle when flowing through the detection region of the flow chamber 133 is reflected by the first dichroic mirror 135 and captured by the side scattering light detector 136, and another portion of the side light passes through the first dichroic mirror 135 to the second dichroic mirror 137, which is also arranged at an angle of 45° to the optical axis downstream of the first dichroic mirror 135. A portion of the side light that passes through the first dichroic mirror 135 is reflected by the second dichroic mirror 137 and captured by the first fluorescence detector 138, and another portion passes through the second dichroic mirror 137 and is captured by the second fluorescence detector 139.

[0112] In some embodiments, the first wavelength is between 315 nm and 490 nm and the second wavelength is between 610 nm and 750 nm. Preferably, the first wavelength is between 400 nm and 450 nm and the second wavelength is between 620 nm and 700 nm. That is, the light of the first wavelength includes blue-violet light, and the light of the second wavelength includes red light.

[0113] In some embodiments, the processor 140 is configured to process and calculate data to obtain a required result. For example, a two-dimensional scatter plot or a three-dimensional scatter plot can be generated according to the collected optical information, and particle analysis can be performed on the scatter plot according to a gating method.

[0114] In some embodiments, the processor 140 can perform visualization processing on an intermediate processing result or a final processing result, and then display the result through the display device 150. For example, the display device 150 can include a user interface, and the processor 140 can output the processing result to the user interface of the display device 150.

[0115] In some embodiments, the processor 140 includes, but is not limited to, a central processing unit (CPU), a micro controller unit (MCU), a field-programmable gate array (FPGA), a digital signal processor (DSP), and the like, which are used to interpret computer instructions and process data in computer software. For example, the processor is used to execute various computer applications in the computer readable storage medium, so that the sample analyzer 100 performs corresponding detection processes and analyzes the scattered light signals and fluorescent light signals detected by the optical detection device 130 in real time.

[0116] In addition, the sample analyzer 100 can further include a liquid path system (not shown) for connecting the sample suction device 110, the sample preparation device 120, and the optical detection device 130, so as to transport liquid among these devices.

[0117] In addition, the sample analyzer 100 can further include a first housing 160 and a second housing 170. The optical detection device 130 and the processor 140 are arranged inside the second housing 170. The sample preparation device 120 is arranged inside the first housing 160, for example, and the display device 150 is arranged on the outer surface of the first housing 160 and is used to display the detection results of the sample analyzer 100.

[0118] In the first embodiment of the present application, the sample suction device 110 is configured to suction a blood sample to be tested; the sample preparation device 120 is configured to mix at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye and a second fluorescent dye in a reaction cell to prepare a test sample; the optical detection device 130 includes a light emitting device, a flow chamber and a light detector, the light emitting device is configured to emit excitation light to irradiate the flow chamber, the flow chamber is in communication with the reaction cell and is configured to pass each particle in the test sample therethrough, the light detector is configured to detect optical information generated after each particle in the test sample passes through the flow chamber and is irradiated by the excitation light in a single test, the light emitting device includes a first light source and a second light source, the first light source is configured to emit light of a first wavelength capable of exciting the first fluorescent dye, the second light source is configured to emit light of a second wavelength capable of exciting the second fluorescent dye, wherein the second wavelength is greater than the first wavelength, and the optical information includes at least one scattering light signal, a first fluorescent signal corresponding to the first fluorescent dye and a second fluorescent signal corresponding to the second fluorescent dye detected in a single test. Here, the processor 140 is configured to obtain infected red blood cell information of the test sample based on the at least one scattering light signal and the first fluorescent signal detected in a single test of the test sample prepared by mixing the blood sample to be tested, the hemolytic agent, the first fluorescent dye and the second fluorescent dye; and obtain a white blood cell four classification result of the test sample based on the at least one scattering light signal and the second fluorescent signal detected in the single test, wherein the white blood cell four classification result includes a count result of a lymphocyte group (lym), a count result of a monocyte group (mon), a count result of a neutrophil group (neu) and a count result of an eosinophil group (eos) in the test sample. Here, the infected red blood cell information refers to optical information related to infected red blood cells.

[0119] Thus, the white blood cell four classification result and the infected red blood cell information can be obtained through one reaction and detection channel, i.e., the malaria-infected red blood cells are specifically detected while the white blood cell classification is realized, which can save the detection time and the blood amount for detection, and realize efficient diagnosis of the disease of the subject (e.g., a fever patient).

[0120] In some embodiments, the scattered light signals include: a first forward scattered light signal and a first side scattered light signal of the particles passing through the flow chamber detected in a single test after being irradiated by light of a first wavelength, and / or a second forward scattered light signal and a second side scattered light signal of the particles passing through the flow chamber detected in a single test after being irradiated by light of a second wavelength. Wherein, the processor 140 obtains the infected red blood cell information of the test sample based on at least one of the scattered light signals and the first fluorescent signal, including: generating a first scatter plot based on one of the first forward scattered light signal, the first side scattered light signal, the second forward scattered light signal, the second side scattered light signal, and the first fluorescent signal, and obtaining the infected red blood cell information of the test sample based on the first scatter plot.

[0121] In some embodiments, the infected red blood cell information can include a count of infected red blood cells. By obtaining a quantitative result of infected red blood cells, the doctor can be more accurately assisted in diagnosing malaria.

[0122] In other embodiments, the infected red blood cell information can include the type of Plasmodium infected by the red blood cells. By typing the Plasmodium infected by the infected red blood cells, the doctor can be assisted in diagnosing malaria for effective treatment.

[0123] In some embodiments, the processor 140 obtains the infected red blood cell information of the test sample based on the first scatter plot, including:

[0124] obtaining a count result of infected red blood cells in the test sample based on the first scatter plot; and / or

[0125] judging the type of Plasmodium infected by the infected red blood cells in the test sample based on the first scatter plot.

[0126] In a specific example, the first scatter plot is generated based on the first forward scattered light signal and the first fluorescent signal, and a region representing infected red blood cells is obtained based on the first scatter plot using a gating technique. The scatter points falling into the region are counted to obtain a count result of infected red blood cells, i.e., a count value. If the count value of infected red blood cells is greater than a predetermined threshold, an alarm prompt is output (judging that the blood sample is a malaria positive sample).

[0127] In another specific example, the first scatter plot is generated according to the first forward scattering light signal and the first fluorescence signal, and a region representing infected red blood cells is obtained from the first scatter plot using a gating technique, and a first group of infected red blood cells and a second group of infected red blood cells are identified from the region, each infected red blood cell in the second group of infected red blood cells containing multiple Plasmodium ring bodies. The center of gravity of the first fluorescence signal and the center of gravity of the first forward scattering light signal of the second group of infected red blood cells are greater than the center of gravity of the first fluorescence signal and the center of gravity of the first forward scattering light signal of the first group of infected red blood cells, respectively. Thus, the count value of infected red blood cells can be more accurately obtained, and a single Plasmodium ring body in multiple Plasmodium ring bodies in an infected red blood cell is avoided from being identified as an infected red blood cell.

[0128] In some embodiments, the processor 140 generates the first scatter plot based on one of the first forward scattering light signal, the first side scattering light signal, the second forward scattering light signal, the second side scattering light signal, and the first fluorescence signal, including: generating the first scatter plot based on the first forward scattering light signal and the first fluorescence signal or based on the second forward scattering light signal and the first fluorescence signal. Here, the processor 140 determines the type of Plasmodium infected by the infected red blood cells in the test sample based on the first scatter plot, including: identifying an infected red blood cell region containing infected red blood cell groups from the first scatter plot, and determining whether the infected red blood cells in the test sample are infected by Plasmodium falciparum, Plasmodium ovale, or Plasmodium vivax based on the forward scattering light intensity of the infected red blood cell groups in the infected red blood cell region.

[0129] In one specific example, the first scatter plot is generated according to the first forward scattering light signal and the first fluorescence signal, and a region representing infected red blood cells is obtained from the first scatter plot using a gating technique, and the type of Plasmodium infected by the infected red blood cells is determined by the size of the center of gravity or the distribution peak of the first forward scattering light signal in the region. Since the sizes of different types of Plasmodium differ, the intensities of the first forward scattering light signals of red blood cells infected with Plasmodium falciparum, Plasmodium ovale, and Plasmodium vivax differ, wherein the center of gravity of the first forward scattering light signal of the red blood cell group infected with Plasmodium vivax is greater than the center of gravity of the first forward scattering light signal of the red blood cell group infected with Plasmodium ovale, and the center of gravity of the first forward scattering light signal of the red blood cell group infected with Plasmodium ovale is greater than the center of gravity of the first forward scattering light signal of the red blood cell group infected with Plasmodium falciparum.

[0130] In some embodiments, the scattered light signals include: a first side scatter light signal detected in a single test for a particle passing through the flow cell after being irradiated by light of the first wavelength, and / or a second side scatter light signal detected in a single test for a particle passing through the flow cell after being irradiated by light of the second wavelength. Here, the processor 140 obtains the white blood cell four-classification result of the test sample based on the at least one scattered light signal and the second fluorescent light signal, including: generating a second scatter plot based on the first side scatter light signal and the second fluorescent light signal or based on the second side scatter light signal and the second fluorescent light signal, and obtaining the white blood cell four-classification result of the test sample based on the second scatter plot.

[0131] In the second embodiment of the present application, the sample suction device 110 is used to suck the blood sample to be tested; the sample preparation device 120 is used to mix at least part of the blood sample to be tested, a hemolytic agent, a first fluorescent dye, a second fluorescent dye and a third fluorescent dye in a reaction cell to prepare a test sample. The optical detection device 130 includes a light emitting device, a flow cell and a light detector, the light emitting device is used to emit excitation light to irradiate the flow cell, the flow cell is in communication with the reaction cell and is used for each particle in the test sample to pass through, and the light detector is used to detect optical information generated after each particle in the test sample is irradiated by the excitation light when passing through the flow cell in a single test. The light emitting device includes a first light source and a second light source, the first light source is used to emit light of a first wavelength capable of exciting the first fluorescent dye, and the second light source is used to emit light of a second wavelength capable of exciting the third fluorescent dye, and the light of the first wavelength or the light of the second wavelength can also excite the second fluorescent dye, wherein the second wavelength is greater than the first wavelength, and the optical information includes at least one scattered light signal detected in a single test, a first fluorescent light signal corresponding to the first fluorescent dye, a second fluorescent light signal corresponding to the second fluorescent dye and a third fluorescent light signal corresponding to the third fluorescent dye. Here, the processor 140 is configured to obtain the infected red blood cell information and the white blood cell five-classification result of the test sample based on the at least one scattered light signal, the first fluorescent light signal, the second fluorescent light signal and the third fluorescent light signal detected in a single test of the test sample prepared by mixing the blood sample to be tested, the hemolytic agent, the first fluorescent dye, the second fluorescent light dye and the third fluorescent dye, wherein the white blood cell five-classification result includes the count result of the lymphocyte group (lym), the count result of the monocyte group (mon), the count result of the neutrophil group (neu), the count result of the eosinophil group (eos) and the count result of the basophil group (baso) in the test sample.

[0132] In some embodiments, the infected red blood cell information can include the count of infected red blood cells and / or the type of Plasmodium infected by the infected red blood cells.

[0133] In some embodiments, the processor 140 obtains the infected red blood cell information and the white blood cell five-classification result of the test sample based on the at least one scattered light signal, the first fluorescent signal, the second fluorescent signal, and the third fluorescent signal, including: obtaining the infected red blood cell information based on one of the at least one scattered light signal, the second fluorescent signal, and the third fluorescent signal, and the first fluorescent signal; and obtaining the white blood cell five-classification result based on the at least one scattered light signal, the second fluorescent signal, and the third fluorescent signal.

[0134] In some embodiments, the processor 140 obtains the infected red blood cell information based on one of the at least one scattered light signal, the second fluorescent signal, and the third fluorescent signal, and the first fluorescent signal, including: generating a first scatter plot based on the at least one scattered light signal and the first fluorescent signal, and obtaining the infected red blood cell information based on the first scatter plot. The processor 140 obtains the white blood cell five-classification result based on the at least one scattered light signal, the second fluorescent signal, and the third fluorescent signal, including: generating a second scatter plot based on the at least one scattered light signal and the second fluorescent signal, obtaining a white blood cell four-classification result of the test sample based on the second scatter plot, wherein the white blood cell four-classification result includes a count result of a lymphocyte group, a count result of a monocyte group, a count result of a neutrophil group, and a count result of an eosinophil group in the test sample; and dividing a first characteristic cell group including a basophil group in the second scatter plot, and obtaining the count result of the basophil group in the test sample based on the at least third fluorescent signal of the first characteristic cell group.

[0135] In some embodiments, the processor 140 obtains the infected red blood cell information based on one of the at least one scattered light signal, the second fluorescent signal, and the third fluorescent signal, and the first fluorescent signal, including: generating a first scatter plot based on the at least one scattered light signal and the first fluorescent signal, and obtaining the infected red blood cell information based on the first scatter plot. The processor 140 obtains the white blood cell five-classification result based on the at least one scattered light signal, the second fluorescent signal, and the third fluorescent signal, including: generating a second scatter plot based on the at least one scattered light signal, for example, a side scattered light signal, and the second fluorescent signal, and obtaining a white blood cell four-classification result of the test sample based on the second scatter plot; and generating a third scatter plot based on the at least one scattered light signal, for example, a side scattered light signal, and the third fluorescent signal, and obtaining the count result of the basophil group in the test sample based on the second scatter plot and the third scatter plot.

[0136] The inventors of the present application have found that the nucleated red blood cells and basophils can be accurately distinguished by the scattered light signal, the second fluorescent signal, and the third fluorescent signal. That is, the processor 140 can distinguish the basophil group and the nucleated red blood cell group in the test sample based on the at least one scattered light signal, the second fluorescent signal, and the third fluorescent signal, to identify and count the basophil group in the test sample, thereby obtaining the count result of the basophil group in the test sample.

[0137] As some implementations, the processor 140 obtains the count result of the basophil group in the test sample based on the second scatter plot and the third scatter plot, including: dividing a first characteristic cell group including the basophil group and the nucleated red blood cell group in the second scatter plot and obtaining a first cell count of the first characteristic cell group; dividing a second characteristic cell group including only the nucleated red blood cell group in the third scatter plot and obtaining a second cell count of the second characteristic cell group; and subtracting the second cell count from the first cell count to obtain the count result of the basophil group in the test sample. That is, the first cell count is the count of basophils and nucleated red blood cells in the test sample, and the second cell count is the count of nucleated red blood cells in the test sample. Here, the second cell count is subtracted from the first cell count, that is, the count result of the basophil group in the test sample is obtained.

[0138] Here, the second cell count is the count value of the nucleated red blood cell group in the test sample.

[0139] Therefore, according to the embodiments of the present application, the basophils and nucleated red blood cells in the test sample can be accurately distinguished, and the infected red blood cell information, the white blood cell five classification result and the nucleated red blood cell information of the test sample can be obtained through one reaction and detection channel.

[0140] In some embodiments, the processor 140 is further configured to obtain the nucleated red blood cell information of the test sample based on at least the third fluorescent signal. In this way, the infected red blood cell information, the white blood cell five classification result and the nucleated red blood cell information can be obtained in one test, greatly reducing the detection cost.

[0141] In other embodiments, the processor 140 is further configured to: generate a first scatter plot based on at least one scattering light signal and the first fluorescent signal, and obtain the infected red blood cell information based on the first scatter plot; generate a second scatter plot based on at least one scattering light signal and the second fluorescent signal, and generate a third scatter plot based on at least one scattering light signal and the third fluorescent signal; obtain the white blood cell five classification result based on the second scatter plot and the third scatter plot; and obtain the nucleated red blood cell information based on the third scatter plot.

[0142] In one specific example, the white blood cell five-classification result is obtained based on the second scatter plot and the third scatter plot, including: obtaining the white blood cell four-classification result of the assay sample based on the second scatter plot, wherein the white blood cell four-classification result includes the count result of the lymphocyte group, the count result of the monocyte group, the count result of the neutrophil group, and the count result of the eosinophil group in the assay sample; dividing the first characteristic cell group including the basophil group and the nucleated red blood cell group in the second scatter plot and obtaining the first cell count of the first characteristic cell group; dividing the second characteristic cell group including only the nucleated red blood cell group in the third scatter plot and obtaining the second cell count of the second characteristic cell group; and subtracting the second cell count from the first cell count to obtain the count result of the basophil group in the assay sample.

[0143] Here, the second cell count is the count value of the nucleated red blood cell group in the assay sample.

[0144] In the third embodiment of the present application, the sample suction device 110 is used to suck the blood sample to be tested; the sample preparation device 120 is used to mix at least part of the blood sample to be tested, a hemolytic agent, a first fluorescent dye, and a third fluorescent dye in a reaction pool to prepare an assay sample; the optical detection device 130 includes a light emitting device, a flow chamber, and a light detector, the light emitting device is used to emit excitation light to irradiate the flow chamber, the flow chamber is in communication with the reaction pool and is used for the passage of each particle in the assay sample, and the light detector is used to detect the optical information generated after each particle in the assay sample is irradiated by the excitation light when passing through the flow chamber in a single test, and the optical information includes at least one scattering light signal, a first fluorescent signal corresponding to the first fluorescent dye, and a third fluorescent signal corresponding to the third fluorescent dye detected in a single test. Here, the processor 140 is configured to obtain the infected red blood cell information and the nucleated red blood cell information of the assay sample based on the at least one scattering light signal, the first fluorescent signal, and the third fluorescent signal detected in a single test of the assay sample prepared by mixing the blood sample to be tested, the hemolytic agent, the first fluorescent dye, and the third fluorescent dye.

[0145] In some embodiments, the first fluorescent dye can be a fluorescent dye capable of specifically labeling Plasmodium, and the third fluorescent dye can be a fluorescent dye capable of identifying nucleated red blood cells.

[0146] In some embodiments, the processor 140 obtains the infected red blood cell information and the nucleated red blood cell information of the assay sample based on the at least one scattering light signal, the first fluorescent signal, and the third fluorescent signal, including: obtaining the count result of the nucleated red blood cell group in the assay sample based on the at least one scattering light signal and the third fluorescent signal; and obtaining the count result of the infected red blood cell and / or the type of Plasmodium infected by the infected red blood cell in the assay sample based on the at least one scattering light signal and the first fluorescent signal.

[0147] In some embodiments, the processor 140 is further configured to obtain a count result of the white blood cells in the assay sample based on the at least one scattered light signal and the third fluorescent signal.

[0148] The present application also provides a sample analysis method 200, as shown in FIG. 3, comprising S210, S220, S230 and S240.

[0149] In step S210, a blood sample to be tested is aspirated. For example, a portion of the blood sample to be tested is aspirated from a test tube containing the blood sample to be tested.

[0150] In step S220, at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye and a second fluorescent dye are mixed to prepare one assay sample at a time.

[0151] For example, in step S220, the hemolytic agent, the first fluorescent dye and the second fluorescent dye can be sequentially added to the same portion of the blood sample to be tested to obtain an assay sample, and then the assay sample is incubated to allow the dyes to sufficiently stain the particles in the assay sample. For another example, in step S220, the first fluorescent dye and the second fluorescent dye can be first mixed with the hemolytic agent to obtain a mixed reagent, and then the mixed reagent is mixed with the blood sample to be tested at a volume ratio of 250:1-1000:1, and after uniform mixing, the mixed assay sample is incubated at a temperature of 25-50°C for 10s-1min, preferably for 10s-40s.

[0152] The various embodiments of the hemolytic agent and the fluorescent dyes used in step S220 can refer to the above description, which will not be repeated here.

[0153] In step S230, each particle in the assay sample is made to pass through an optical detection area irradiated by excitation light in a single test to obtain optical information generated by each particle in the assay sample after being irradiated by the excitation light, the excitation light including light of a first wavelength capable of exciting the first fluorescent dye and light of a second wavelength capable of exciting the second fluorescent dye, wherein the second wavelength is greater than the first wavelength, and the optical information includes at least one scattered light signal, a first fluorescent signal corresponding to the first fluorescent dye and a second fluorescent signal corresponding to the second fluorescent dye obtained in a single test.

[0154] Here, the first fluorescent signal is a fluorescent signal generated by the particles in the assay sample after being combined with the first fluorescent dye under excitation of light of the first wavelength, and the second fluorescent signal is a fluorescent signal generated by the particles in the assay sample after being combined with the second fluorescent dye under excitation of light of the second wavelength.

[0155] That is, in step S230, the scattering light signal (or scattering light signal intensity) and the fluorescent light signal (or fluorescent light signal intensity) of the assay sample are obtained based on the principle of flow cytometry.

[0156] In step S240, the infected red blood cell information of the assay sample is obtained based on the at least one scattering light signal and the first fluorescent light signal; and the white blood cell four-classification result of the assay sample is obtained based on the at least one scattering light signal and the second fluorescent light signal, wherein the white blood cell four-classification result includes the count result of the lymphocyte group, the count result of the monocyte group, the count result of the neutrophil group and the count result of the eosinophil group in the assay sample.

[0157] According to another aspect of the present application, a sample analysis method 300 is provided, as shown in FIG. 4, which includes steps S310, S320, S330 and S340.

[0158] In step S310, the blood sample to be tested is aspirated. For example, a portion of the blood sample to be tested is aspirated from a test tube containing the blood sample to be tested.

[0159] In step S320, at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye, a second fluorescent dye and a third fluorescent dye are mixed to prepare one assay sample at a time.

[0160] For example, in step S320, the hemolytic agent, the first fluorescent dye, the second fluorescent dye and the third fluorescent dye can be sequentially added to the same portion of the blood sample to be tested to obtain an assay sample, and then the assay sample is incubated to allow the dyes to sufficiently stain the particles in the assay sample. For another example, in step S320, the first fluorescent dye, the second fluorescent dye and the third fluorescent dye can be first mixed with the hemolytic agent to obtain a mixed reagent, and then the mixed reagent is mixed with the blood sample to be tested at a volume ratio of 250:1-1000:1, and after being mixed uniformly, the obtained assay sample is incubated at a temperature of 25-50°C for 10s-1min, preferably for 10s-40s.

[0161] The various embodiments of the hemolytic agent and the fluorescent dyes used in step S320 can refer to the above description, which will not be repeated here.

[0162] In step S330, each particle in the assay sample is caused to pass through the optical detection region irradiated by the excitation light in a single test to obtain optical information generated by each particle in the assay sample after being irradiated by the excitation light, the excitation light including light of a first wavelength capable of exciting the first fluorescent dye and light of a second wavelength capable of exciting the third fluorescent dye, the light of the first wavelength or the light of the second wavelength also being capable of exciting the second fluorescent dye, wherein the second wavelength is greater than the first wavelength, the optical information including at least one scattering light signal, a first fluorescent signal corresponding to the first fluorescent dye, a second fluorescent signal corresponding to the second fluorescent dye, and a third fluorescent signal corresponding to the third fluorescent dye, which are obtained in a single test.

[0163] Here, the first fluorescent signal is a fluorescent signal generated by the particle in the assay sample after being combined with the first fluorescent dye under excitation of the light of the first wavelength, the second fluorescent signal is a fluorescent signal generated by the particle in the assay sample after being combined with the second fluorescent dye under excitation of the light of the first wavelength, and the third fluorescent signal is a fluorescent signal generated by the particle in the assay sample after being combined with the third fluorescent dye under excitation of the light of the second wavelength.

[0164] That is, in step S330, the scattering light signal (or the scattering light signal intensity) and the fluorescent signal (or the fluorescent signal intensity) of the assay sample are obtained based on the principle of flow cytometry.

[0165] In step S340, the information of infected red blood cells and the five classification results of white blood cells of the assay sample are obtained based on the at least one scattering light signal, the first fluorescent signal, the second fluorescent signal, and the third fluorescent signal, wherein the five classification results of white blood cells include the count result of the lymphocyte group, the count result of the monocyte group, the count result of the neutrophil group, the count result of the eosinophil group, and the count result of the basophil group in the assay sample.

[0166] According to still another aspect of the present application, a sample analysis method 400 is provided, as shown in FIG. 5, including S410, S420, S430, and S440. It includes:

[0167] In step S410, a blood sample to be tested is aspirated. For example, a portion of the blood sample to be tested is aspirated from a test tube containing the blood sample to be tested.

[0168] In step S420, at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye, and a third fluorescent dye are mixed to prepare one assay sample in a single test.

[0169] For example, in step S420, the hemolytic agent, the first fluorescent dye and the third fluorescent dye can be added into the same portion of the blood sample to be tested in sequence to obtain a test sample, and then the test sample is incubated so that the dyes can sufficiently dye the particles in the test sample. For another example, in step S420, the first fluorescent dye and the third fluorescent dye can be mixed with the hemolytic agent in advance to obtain a mixed reagent, and then the mixed reagent can be mixed with the blood sample to be tested at a volume ratio of 250:1-1000:1, and after being mixed uniformly, the mixed test sample is incubated at a temperature of 25-50°C for 10s-1min, preferably for 10s-40s.

[0170] The various embodiments of the hemolytic agent and the fluorescent dyes used in step S420 can refer to the above description, which will not be repeated here.

[0171] In step S430, each particle in the test sample is made to pass through the optical detection area irradiated by the excitation light in a single test to obtain optical information generated after each particle in the test sample is irradiated by the excitation light, and the optical information includes at least one scattering light signal, a first fluorescent signal corresponding to the first fluorescent dye and a third fluorescent signal corresponding to the third fluorescent dye obtained in a single test.

[0172] Here, the first fluorescent signal is a fluorescent signal generated after the particles in the test sample are combined with the first fluorescent dye under excitation of light at the first wavelength, and the third fluorescent signal is a fluorescent signal generated after the particles in the test sample are combined with the third fluorescent dye under excitation of light at the second wavelength.

[0173] That is, in step S430, the scattering light signal (or the scattering light signal intensity) and the fluorescent signal (or the fluorescent signal intensity) of the test sample are obtained based on the principle of flow cytometry.

[0174] In step S440, the information of the infected red blood cells and the information of the nucleated red blood cells of the test sample are obtained based on the at least one scattering light signal, the first fluorescent signal and the third fluorescent signal.

[0175] The present application will now be described with reference to the following examples, which are intended to illustrate the present application and not to limit the present application. Unless specifically indicated otherwise, the experiments and methods described in the examples are carried out substantially in accordance with the conventional methods well known in the art and described in various references. In addition, unless specific conditions are indicated in the examples, the conventional conditions or the conditions recommended by the manufacturer are used. Unless the manufacturer of the reagent or instrument is indicated, it is a conventional product that can be obtained by purchase on the market.

[0176] Example 1

[0177] The reagent formula is as follows:

[0178] A: fluorescent dye solution

[0179] First fluorescent dye 30mg

[0180] Second fluorescent dye 50mg

[0181] Ethylene glycol 1000g

[0182] B: hemolytic agent

[0183] The first fluorescent dye has the following general formula:

[0184] The second fluorescent dye has the following general formula:

[0185] The sample analyzer according to the present application is equipped with a red laser (emitting a light beam with a wavelength of about 633 nm) and a blue laser (emitting a light beam with a wavelength of about 488 nm). The sample analyzer is used to process and detect a blood sample not infected with Plasmodium and a blood sample infected with Plasmodium. Specifically, 20 microliters of reagent A and 20 microliters of the blood sample are added to 1 mL of reagent B to form a test sample. The intensity of scattered light corresponding to blue light excitation, the first fluorescent intensity corresponding to blue light excitation, and the second fluorescent intensity corresponding to red light excitation are detected. A two-dimensional first scatter plot is generated based on the first fluorescent intensity and the intensity of scattered light (forward scattered light intensity or side scattered light intensity). The region representing infected red blood cells, Infect_Area, is obtained from the first scatter plot. The count of infected red blood cells is obtained by counting the scatter points falling into the region. A two-dimensional second scatter plot is generated based on the second fluorescent intensity and the side scattered light intensity. The white blood cell four-classification result is obtained based on the second scatter plot. The white blood cell four-classification result includes the count result of the neutrophil group (neu), the count result of the lymphocyte group (lym), the count result of the monocyte group (mon), and the count result of the eosinophil group (eos) in the test sample.

[0186] FIGS. 6a and 6b show the first scatter plot obtained by processing and detecting the blood sample not infected with Plasmodium using the sample analyzer, and FIG. 6c shows the second scatter plot obtained by processing and detecting the blood sample not infected with Plasmodium using the sample analyzer. There are basically no scatter points in the infected red blood cell region Infect_Area of the first scatter plot of FIG. 6, indicating that the blood sample is not infected with Plasmodium.

[0187] Fig. 7a and Fig. 7b show the first scatter plots obtained by processing and detecting the blood sample to be tested infected with Plasmodium by the sample analyzer, and Fig. 7c shows the second scatter plot obtained by processing and detecting the blood sample to be tested infected with Plasmodium by the sample analyzer. There are a certain number of scatter points in the infected red blood cell area Infect_Area of the first scatter plot of Fig. 7, indicating that the blood sample to be tested is infected with Plasmodium.

[0188] Example 2

[0189] The blood samples to be tested infected with different types of Plasmodium were processed and detected by the sample analyzer according to the present application equipped with a red laser (emitting a light beam with a wavelength of about 633 nm) and a blue laser (emitting a light beam with a wavelength of about 488 nm) respectively: 20 microliters of the above-mentioned reagent A (reagent A of Example 1) and 20 microliters of the blood sample to be tested were added to 1 mL of the above-mentioned reagent B (reagent B of Example 1) to form a test sample; the forward scattering light intensity corresponding to blue light excitation, the first fluorescence intensity corresponding to blue light excitation, and the second fluorescence intensity corresponding to red light excitation were detected; a two-dimensional first scatter plot was generated based on the first fluorescence intensity and the forward scattering light intensity, and the area representing the infected red blood cells Infect_Area was obtained according to the first scatter plot, and by identifying the range where the distribution peak of the forward scattering light intensity in the area is located, the type of Plasmodium infected by the infected red blood cells can be determined.

[0190] Fig. 8a shows the first scatter plot obtained by processing and detecting the blood sample to be tested infected with Plasmodium falciparum by the sample analyzer, Fig. 8b shows the first scatter plot obtained by processing and detecting the blood sample to be tested infected with Plasmodium ovale by the sample analyzer, and Fig. 8c shows the first scatter plot obtained by processing and detecting the blood sample to be tested infected with Plasmodium vivax by the sample analyzer. Since the sizes of the bodies of different types of Plasmodium are different, the intensities of the forward scattering light signals of the red blood cells infected with Plasmodium falciparum, Plasmodium ovale and Plasmodium vivax are different, wherein the center of gravity of the forward scattering light intensity of the red blood cell population infected with Plasmodium vivax is greater than that of the red blood cell population infected with Plasmodium ovale, and the center of gravity of the forward scattering light of the red blood cell population infected with Plasmodium ovale is greater than that of the red blood cell population infected with Plasmodium falciparum.

[0191] Example 3

[0192] The sample analyzer according to the present application equipped with a red laser (emitting a light beam having a wavelength of about 633 nm) and a blue laser (emitting a light beam having a wavelength of about 488 nm) is used to process and detect a blood sample to be tested which is not infected with Plasmodium and a blood sample to be tested which is infected with Plasmodium: 20 microliters of the above-described reagent A (reagent A of Example 1) and 20 microliters of the blood sample to be tested are added to 1 mL of the above-described reagent B (reagent B of Example 1) to form an assay sample; the intensity of scattered light corresponding to excitation by blue light, the first fluorescence intensity corresponding to excitation by blue light, and the second fluorescence intensity corresponding to excitation by red light are detected.

[0193] A two-dimensional first scatter plot is generated based on the first fluorescence intensity and the intensity of scattered light (forward scattered light intensity or side scattered light intensity), as shown in FIGS. 9a and 9b, and an area representing infected red blood cells, Infect_Area, is obtained from the first scatter plot, and the scatter points falling within the area are counted to obtain a count value of infected red blood cells; and a two-dimensional second scatter plot is generated based on the second fluorescence intensity and the side scattered light intensity, as shown in FIG. 9c, and a white blood cell four-classification result is obtained based on the second scatter plot, the white blood cell four-classification result including a count result of a neutrophil group (neu), a count result of a lymphocyte group (lym), a count result of a monocyte group (mon), and a count result of an eosinophil group (eos) in the assay sample.

[0194] Here, in the first scatter plot shown in FIGS. 9a and 9b, an area representing infected red blood cells, Infect_Area, is divided, and a first infected red blood cell group Infect_CellPop1 and a second infected red blood cell group Infect_CellPop2 are identified from the area Infect_Area, and each infected red blood cell in the second infected red blood cell group Infect_CellPop2 contains a plurality of Plasmodium ring bodies.

[0195] Example 4

[0196] The reagent formulations are as follows:

[0197] A: fluorescent dye

[0198] B: hemolytic agent

[0199] The first fluorescent dye has the following general formula:

[0200] The second fluorescent dye has the following general formula:

[0201] The third fluorescent dye has the following general formula:

[0202] The sample analyzer equipped with a red laser (emitting a light beam with a wavelength of about 633 nm) and a blue laser (emitting a light beam with a wavelength of about 488 nm) according to the present application is used to process and detect a blood sample to be tested infected with Plasmodium: 20 microliters of reagent A and 20 microliters of the blood sample to be tested are added into 1 mL of reagent B to form a test sample; the forward scattering light intensity and the side scattering light intensity corresponding to blue light excitation, the first fluorescence intensity and the second fluorescence intensity corresponding to blue light excitation, and the third fluorescence intensity corresponding to red light excitation are detected.

[0203] A two-dimensional first scatter plot is generated based on the first fluorescence intensity and the scattering light intensity (forward scattering light intensity or side scattering light intensity), as shown in FIGS. 10a and 10b, and an area representing infected red blood cells, Infect_Area, is obtained from the first scatter plot, and the scatter points falling into the area are counted to obtain the count value of infected red blood cells; a two-dimensional second scatter plot is generated based on the second fluorescence intensity and the side scattering light intensity, as shown in FIG. 10c; and a two-dimensional third scatter plot is generated based on the third fluorescence intensity and the side scattering light intensity, as shown in FIG. 10d. Here, the white blood cell five-classification result is obtained based on the second scatter plot and the third scatter plot, and the white blood cell five-classification result includes the count result of the neutrophil group (neu), the count result of the lymphocyte group (lym), the count result of the monocyte group (mon), the count result of the eosinophil group (eos), and the count result of the basophil group (baso) of the test sample, and the count result of the nucleated red blood cell group (NRBC) is obtained based on the third scatter plot.

[0204] Example 5

[0205] The reagent formula is as follows:

[0206] A: fluorescent dye

[0207] First fluorescent dye 30 mg

[0208] Third fluorescent dye 30 mg

[0209] Ethylene glycol 1000 g

[0210] B: hemolytic agent

[0211] The first fluorescent dye has the following general formula:

[0212] The third fluorescent dye has the following general formula:

[0213] The sample analyzer according to the present application is equipped with a red laser (emitting a light beam with a wavelength of about 633 nm) and a blue laser (emitting a light beam with a wavelength of about 488 nm). A blood sample infected with Plasmodium is processed and detected as follows: 20 microliters of reagent A and 20 microliters of the blood sample are added to 1 mL of reagent B to form a test sample; the forward scattering light intensity and the side scattering light intensity corresponding to blue light excitation, the first fluorescence intensity corresponding to blue light excitation, and the second fluorescence intensity corresponding to red light excitation are detected.

[0214] A two-dimensional first scatter plot is generated based on the first fluorescence intensity and the forward scattering light intensity, as shown in FIG. 11a. An area representing infected red blood cells, Infect_Area, is obtained from the first scatter plot. The count of scatter points falling within the area is obtained to obtain the count of infected red blood cells. A two-dimensional third scatter plot is generated based on the second fluorescence intensity and the side scattering light intensity, as shown in FIG. 11b. The count of nucleated red blood cells (NRBC) is obtained based on the third scatter plot.

[0215] The features or combinations of features mentioned in the specification, drawings and claims of the present application can be used in any combination or alone, as long as they are meaningful and do not contradict each other within the scope of the present application. The advantages and features of the sample analyzer described in the present application apply to the sample analysis method provided by the present application in a corresponding manner, and vice versa.

[0216] The above description is only a preferred embodiment of the present application and does not limit the scope of patent protection of the present application. Any equivalent transformation or direct / indirect application in other related technical fields made by using the content of the present application and the drawings is included in the scope of patent protection of the present application.

Claims

1. A sample analyzer, comprising: a sample drawing device configured to draw a blood sample to be tested; a sample preparation device configured to mix at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye and a second fluorescent dye in a reaction cell to prepare an assay sample; an optical detection device including a light emitting device configured to emit excitation light to illuminate a flow cell, the flow cell being in communication with the reaction cell and configured to pass each particle in the assay sample therethrough, and a light detector configured to detect optical information generated after each particle in the assay sample passes through the flow cell and is illuminated by the excitation light in a single test, the light emitting device including a first light source configured to emit light of a first wavelength capable of exciting the first fluorescent dye and a second light source configured to emit light of a second wavelength capable of exciting the second fluorescent dye, wherein the second wavelength is greater than the first wavelength, and the optical information includes at least one scatter signal, a first fluorescent signal corresponding to the first fluorescent dye and a second fluorescent signal corresponding to the second fluorescent dye detected in the single test; and a processor configured to obtain infected red blood cell information of the assay sample based on at least one of the scatter signals and the first fluorescent signal, and obtain a white blood cell differential count result of the assay sample based on at least one of the scatter signals and the second fluorescent signal, wherein the white blood cell differential count result includes a count result of a lymphocyte population, a count result of a monocyte population, a count result of a neutrophil population and a count result of an eosinophil population in the assay sample.

2. The sample analyzer of claim 1, wherein, The first wavelength is between 315 nm and 490 nm and the second wavelength is between 610 nm and 750 nm.

3. The sample analyzer of claim 1 or 2, wherein, The scatter signals include a first forward scatter signal and a first side scatter signal generated after a particle passing through the flow cell is illuminated by the light of the first wavelength, and / or a second forward scatter signal and a second side scatter signal generated after a particle passing through the flow cell is illuminated by the light of the second wavelength detected in the single test. The processor obtains the infected red blood cell information of the assay sample based on at least one of the scatter signals and the first fluorescent signal, including: generating a first scatter plot based on one of the first forward scatter signal, the first side scatter signal, the second forward scatter signal, the second side scatter signal and the first fluorescent signal, and obtaining the infected red blood cell information of the assay sample based on the first scatter plot.

4. The sample analyzer of claim 3, wherein, The processor obtains the infected red blood cell information of the assay sample based on the first scatter plot, including: obtaining a count result of infected red blood cells in the assay sample based on the first scatter plot; and / or judging a type of Plasmodium infecting the infected red blood cells in the assay sample based on the first scatter plot.

5. The sample analyzer of claim 4, wherein, The processor generates a first scatter plot based on one of the first forward scattering light signal, the first side scattering light signal, the second forward scattering light signal, the second side scattering light signal and the first fluorescence signal, including generating the first scatter plot based on the first forward scattering light signal and the first fluorescence signal or based on the second forward scattering light signal and the first fluorescence signal; The processor determines the type of malaria parasite infecting the infected red blood cells in the test sample based on the first scatter plot, including identifying an infected red blood cell region containing infected red blood cell groups from the first scatter plot, and determining whether the infected red blood cells in the test sample are infected with Plasmodium falciparum, Plasmodium ovale or Plasmodium vivax based on the forward scattering light intensity of the infected red blood cell groups in the infected red blood cell region.

6. The sample analyzer of any one of claims 1 to 5, wherein, The scattering light signals include a first side scattering light signal generated by a particle passing through the flow chamber after being irradiated by light of the first wavelength detected in the single test, and / or a second side scattering light signal generated by a particle passing through the flow chamber after being irradiated by light of the second wavelength detected in the single test; The processor obtains the white blood cell four-classification result of the test sample based on at least one of the scattering light signals and the second fluorescence signal, including: The processor generates a second scatter plot based on the first side scattering light signal and the second fluorescence signal or based on the second side scattering light signal and the second fluorescence signal, and obtains the white blood cell four-classification result of the test sample based on the second scatter plot.

7. A sample analyzer, comprising: a sample suction device configured to suction a blood sample to be tested; a sample preparation device configured to mix at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye, a second fluorescent dye, and a third fluorescent dye in a reaction cell to prepare a test sample; an optical detection device including a light emitting device, a flow chamber, and a light detector, the light emitting device configured to emit excitation light to irradiate the flow chamber, the flow chamber being in communication with the reaction cell and configured to pass each particle in the test sample therethrough, the light detector configured to detect optical information generated by each particle in the test sample after being irradiated by the excitation light when passing through the flow chamber in a single test, the light emitting device including a first light source configured to emit light of a first wavelength capable of exciting the first fluorescent dye and a second light source configured to emit light of a second wavelength capable of exciting the third fluorescent dye, the light of the first wavelength or the light of the second wavelength also being capable of exciting the second fluorescent dye, wherein the second wavelength is greater than the first wavelength, and the optical information includes at least one scattering light signal detected in the single test, a first fluorescence signal corresponding to the first fluorescent dye, a second fluorescence signal corresponding to the second fluorescent dye, and a third fluorescence signal corresponding to the third fluorescent dye; and The processor is configured to obtain the information of infected red blood cells and the white blood cell five-classification result of the test sample based on at least one of the scattered light signals, the first fluorescence signal, the second fluorescence signal and the third fluorescence signal, wherein the white blood cell five-classification result comprises a count result of a lymphocyte group, a count result of a monocyte group, a count result of a neutrophil group, a count result of an eosinophil group and a count result of a basophil group in the test sample.

8. The sample analyzer of claim 7, wherein, The processor obtains the information of infected red blood cells and the white blood cell five-classification result of the test sample based on at least one of the scattered light signals, the first fluorescence signal, the second fluorescence signal and the third fluorescence signal, comprising: the information of infected red blood cells is obtained based on at least one of the scattered light signals, the second fluorescence signal and the third fluorescence signal; and the white blood cell five-classification result is obtained based on at least one of the scattered light signals, the second fluorescence signal and the third fluorescence signal.

9. The sample analyzer according to claim 8, wherein the processor obtains the information of infected red blood cells based on at least one of the scattered light signals, the second fluorescence signal and the third fluorescence signal, comprising: a first scatter plot is generated based on at least one of the scattered light signals and the first fluorescence signal, and the information of infected red blood cells is obtained based on the first scatter plot; and the processor obtains the white blood cell five-classification result based on at least one of the scattered light signals, the second fluorescence signal and the third fluorescence signal, comprising: a second scatter plot is generated based on at least one of the scattered light signals and the second fluorescence signal, a white blood cell four-classification result of the test sample is obtained based on the second scatter plot, wherein the white blood cell four-classification result comprises a count result of a lymphocyte group, a count result of a monocyte group, a count result of a neutrophil group and a count result of an eosinophil group in the test sample, and a first characteristic cell group comprising a basophil group is divided in the second scatter plot, and a count result of the basophil group in the test sample is obtained based on at least a third fluorescence signal of the first characteristic cell group.

10. The sample analyzer according to claim 8, wherein the processor obtains the information of infected red blood cells based on at least one of the scattered light signals, the second fluorescence signal and the third fluorescence signal, comprising: a first scatter plot is generated based on at least one of the scattered light signals and the first fluorescence signal, and the information of infected red blood cells is obtained based on the first scatter plot; and the processor obtains the white blood cell five-classification result based on at least one of the scattered light signals, the second fluorescence signal and the third fluorescence signal, comprising: generate a second scatter plot based on at least one of the scatter light signals and the second fluorescent light signal, and obtain the white blood cell four-classification result of the test sample based on the second scatter plot, wherein the white blood cell four-classification result comprises a count result of a lymphocyte group, a count result of a monocyte group, a count result of a neutrophil group, and a count result of an eosinophil group in the test sample; and generate a third scatter plot based on at least one of the scatter light signals and the third fluorescent light signal, and obtain the count result of the basophil group in the test sample based on the second scatter plot and the third scatter plot.

11. The sample analyzer of claim 10, wherein, The processor obtains the count result of the basophil group in the test sample based on the second scatter plot and the third scatter plot, comprising: dividing a first characteristic cell group comprising the basophil group and the nucleated red blood cell group in the second scatter plot and obtaining a first cell count of the first characteristic cell group; dividing a second characteristic cell group comprising only the nucleated red blood cell group in the third scatter plot and obtaining a second cell count of the second characteristic cell group; and subtracting the second cell count from the first cell count to obtain the count result of the basophil group in the test sample.

12. The sample analyzer of claim 7 or 8, wherein, The processor is further configured to obtain the nucleated red blood cell information of the test sample based on at least the third fluorescent light signal.

13. The sample analyzer of claim 12, wherein, The processor is further configured to: generate a first scatter plot based on at least one of the scatter light signals and the first fluorescent light signal, and obtain the infected red blood cell information based on the first scatter plot; generate a second scatter plot based on at least one of the scatter light signals and the second fluorescent light signal, and generate a third scatter plot based on at least one of the scatter light signals and the third fluorescent light signal; obtain the white blood cell five-classification result based on the second scatter plot and the third scatter plot; and obtain the nucleated red blood cell information based on the third scatter plot. The obtaining of the white blood cell five-classification result based on the second scatter plot and the third scatter plot comprises:

14. The sample analyzer of claim 13, wherein, obtaining a white blood cell four-classification result of the test sample based on the second scatter plot, wherein the white blood cell four-classification result comprises a count result of a lymphocyte group, a count result of a monocyte group, a count result of a neutrophil group, and a count result of an eosinophil group in the test sample; dividing a first characteristic cell group comprising the basophil group and the nucleated red blood cell group in the second scatter plot and obtaining a first cell count of the first characteristic cell group; dividing a second characteristic cell group comprising only the nucleated red blood cell group in the third scatter plot and obtaining a second cell count of the second characteristic cell group; and subtracting the second cell count from the first cell count to obtain the count result of the basophil group in the test sample.

15. A sample analyzer, comprising: a sample suction device configured to suction a blood sample to be tested; a sample preparation device configured to mix at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye, and a third fluorescent dye in a reaction cell to prepare a test sample; and a sample analysis device configured to analyze the test sample. An optical detection device comprising a light emitting device for emitting excitation light to illuminate a flow cell, a flow chamber in communication with the flow cell and for passing individual particles in the assay sample therethrough, and a light detector for detecting optical information generated by individual particles in the assay sample when illuminated by the excitation light as the individual particles pass through the flow chamber in a single test, the optical information comprising at least one scatter light signal, a first fluorescent signal corresponding to the first fluorescent dye, and a third fluorescent signal corresponding to the third fluorescent dye detected in the single test; and a processor configured to derive infected red blood cell information and nucleated red blood cell information of the assay sample based on at least one of the scatter light signal, the first fluorescent signal, and the third fluorescent signal.

16. The sample analyzer of claim 15, wherein, The light emitting device comprises a first light source for emitting light of a first wavelength capable of exciting the first fluorescent dye, and a second light source for emitting light of a second wavelength capable of exciting the third fluorescent dye, wherein the second wavelength is greater than the first wavelength, preferably the first wavelength is between 315 nm and 490 nm and the second wavelength is between 610 nm and 750 nm; wherein the processor derives infected red blood cell information and nucleated red blood cell information of the assay sample based on at least one of the scatter light signal, the first fluorescent signal, and the third fluorescent signal, comprises: deriving a count result of a nucleated red blood cell population in the assay sample based on at least one of the scatter light signal and the third fluorescent signal; and deriving a count result of infected red blood cells in the assay sample and / or a type of Plasmodium infecting the infected red blood cells based on at least one of the scatter light signal and the first fluorescent signal.

17. The sample analyzer of claim 15 or 16, wherein, The processor is further configured to: derive a count result of white blood cells in the assay sample based on at least one of the scatter light signal and the third fluorescent signal.

18. A sample analysis method comprising: aspirating a blood sample to be tested; mixing at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye, and a second fluorescent dye to prepare an assay sample; passing individual particles in the assay sample through an optical detection region illuminated by excitation light in a single test to obtain optical information generated by individual particles in the assay sample when illuminated by the excitation light, the excitation light comprising light of a first wavelength capable of exciting the first fluorescent dye and light of a second wavelength capable of exciting the second fluorescent dye, wherein the second wavelength is greater than the first wavelength, the optical information comprising at least one scatter light signal, a first fluorescent signal corresponding to the first fluorescent dye, and a second fluorescent signal corresponding to the second fluorescent dye obtained in the single test; deriving infected red blood cell information of the assay sample based on at least one of the scatter light signal and the first fluorescent signal; and obtaining the white blood cell four-classification result of the test sample based on at least one of the scattering light signals and the second fluorescence signal, wherein the white blood cell four-classification result comprises a count result of a lymphocyte group, a count result of a monocyte group, a count result of a neutrophil group, and a count result of an eosinophil group in the test sample. 19.A method of analyzing a sample, comprising: aspirating a blood sample to be tested; mixing at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye, a second fluorescent dye, and a third fluorescent dye to prepare a test sample; passing each particle in the test sample through an optical detection region irradiated by excitation light in a single test to obtain optical information generated after each particle in the test sample is irradiated by the excitation light, the excitation light comprising light of a first wavelength capable of exciting the first fluorescent dye and light of a second wavelength capable of exciting the third fluorescent dye, the light of the first wavelength or the light of the second wavelength also being capable of exciting the second fluorescent dye, wherein the second wavelength is greater than the first wavelength, and the optical information comprises at least one scattering light signal, a first fluorescence signal corresponding to the first fluorescent dye, a second fluorescence signal corresponding to the second fluorescent dye, and a third fluorescence signal corresponding to the third fluorescent dye obtained in the single test; and obtaining infected red blood cell information and a white blood cell five-classification result of the test sample based on at least one of the scattering light signals, the first fluorescence signal, the second fluorescence signal, and the third fluorescence signal, wherein the white blood cell five-classification result comprises a count result of a lymphocyte group, a count result of a monocyte group, a count result of a neutrophil group, a count result of an eosinophil group, and a count result of a basophil group in the test sample. 20.A method of analyzing a sample, comprising: aspirating a blood sample to be tested; mixing at least a portion of the blood sample to be tested, a hemolytic agent, a first fluorescent dye, and a third fluorescent dye to prepare a test sample; passing each particle in the test sample through an optical detection region irradiated by excitation light in a single test to obtain optical information generated after each particle in the test sample is irradiated by the excitation light, the optical information comprising at least one scattering light signal, a first fluorescence signal corresponding to the first fluorescent dye, and a third fluorescence signal corresponding to the third fluorescent dye obtained in the single test; and obtaining infected red blood cell information and nucleated red blood cell information of the test sample based on at least one of the scattering light signals, the first fluorescence signal, and the third fluorescence signal. ​

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