阅读说明：本技术 一种血液检测方法及血液分析系统 (Blood detection method and blood analysis system ) 是由 陈庚文 张子千 叶燚 李朝阳 于 2019-04-26 设计创作，主要内容包括：本发明公开了一种血液检测方法,包括：用第一试剂处理血液样本以获得待测试样,所述第一试剂包括溶血剂,所述溶血剂将所述血液样本中的红细胞裂解为其光散射特性显著不同于血小板的碎片；使待测试样中的粒子逐个通过光学检测系统的检测区,获取所述待测试样的光学信息；和根据所述光学信息中的至少两种获得血小板的光学信息。本发明的方法通过裂解血液样本中的红细胞获取血小板计数,并可同时区分白细胞亚群。(The invention discloses a blood detection method, which comprises the following steps: treating a blood sample with a first reagent to obtain a test sample, the first reagent comprising a hemolytic agent that lyses red blood cells in the blood sample into fragments whose light scattering properties are significantly different from platelets; enabling particles in a sample to be detected to pass through a detection area of an optical detection system one by one, and acquiring optical information of the sample to be detected; and obtaining optical information of the platelets from at least two of the optical information. The method of the invention obtains the platelet count by lysing the red blood cells in the blood sample and can simultaneously distinguish the leukocyte subsets.)

A method of blood testing, the method comprising:

treating a blood sample with a first reagent to obtain a test sample, the first reagent comprising a hemolytic agent that lyses red blood cells in the blood sample into fragments whose light scattering properties are significantly different from platelets;

enabling particles in a sample to be detected to pass through a detection area of an optical detection system one by one, and acquiring optical information of the sample to be detected; and

optical information of the platelets is obtained from at least two of the optical information.

The blood detection method of claim 1, wherein the at least two optical information are a forward scattered light intensity and a side scattered light intensity to distinguish platelets from lysed red blood cell debris.

A blood detection method according to claim 2, the method further comprising obtaining optical information of the leukocytes from the optical information of the forward scattered light intensity and the side scattered light intensity, preferably differentiating the subpopulations of leukocytes according to the obtained optical information of the leukocytes to obtain subpopulations of leukocytes comprising at least monocytes, lymphocytes and neutrophils.

The method for detecting blood according to claim 1, wherein the hemolytic agent is at least one selected from the group consisting of alkylglycoside, triterpenoid saponin, and steroid saponin.

The method for detecting blood according to claim 4, wherein the alkyl glycoside is selected from the group consisting of glycosides having the general formula I:

R-(CH₂)_n-CH₃(I)

wherein R is selected from the group consisting of monosaccharides, deoxymonosaccharides, and polysaccharides, and n is an integer of 5-17.

The blood detection method of claim 1, wherein the first reagent further comprises:

a nonionic surfactant having the general formula II:

R₁-R₂-(CH₂CH₂O)_m-H (II)

wherein R is₁Is C8-C23 alkyl, R₂is-O-),

optionally, at least one organic acid or a salt thereof, wherein the organic acid or salt thereof is selected from the group consisting of organic acids having at least one carboxyl group or sulfonic acid group and alkali metal salts thereof.

The blood detection method of claim 1, further comprising treating the blood sample with a second reagent, the second reagent comprising a fluorescent dye.

The blood detection method according to claim 7, wherein the particles in the test sample are individually passed through a detection region of an optical detection system, the optical information of the test sample is acquired further comprising fluorescence information, and the subpopulation of leukocytes is distinguished according to the side scattered light intensity and the fluorescence intensity information to obtain a subpopulation of leukocytes at least comprising monocytes, lymphocytes and neutrophils and/or to identify immature granulocytes.

The blood detection method of claim 7 or 8, wherein the fluorescent dye comprises a first fluorescent dye selected from a membrane-specific dye and a mitochondrial-specific dye.

The method for detecting blood according to claim 9, wherein the membrane-specific dye is selected from the group consisting of DiA, DiD, DiI, DiO, DiR, DiS, FDA, Alexa Fluor488, Super Fluor488 and modified structures thereof as precursors, and the mitochondrial-specific dye is selected from the group consisting of Janus Green B, Mitolite Red, rhodamine 123 and Mitotricker series and precursors thereof.

The blood detection method according to claim 9 or 10, wherein the platelets are identified based on fluorescence intensity and forward scattered light intensity information.

The blood detection method according to claim 9 or 10, wherein the method further comprises: the reticulocytes are alarmed when the number of particles in a preset area of a scatter diagram formed from the forward scattered light intensity and the fluorescence intensity exceeds a predetermined threshold.

The blood detection method according to claim 7 or 8, wherein the fluorescent dye comprises a second fluorescent dye selected from one of nucleic acid-specific dyes, preferably the nucleic acid-specific dye is a nucleic acid-specific dye for reticulocytes.

The blood detection method of claim 13, wherein the method further comprises: the reticulocytes are identified based on the fluorescence intensity and scattered light intensity information, and preferably, the platelets and reticulocytes are distinguished based on the fluorescence intensity and forward scattered light intensity information.

The blood detection method of claim 14, wherein the method further comprises: reticulocytes were counted according to fluorescence intensity and forward scattered light intensity information.

The blood detection method according to claim 7 or 8, wherein the fluorescent dye includes a first fluorescent dye selected from one of membrane-specific dyes and mitochondrial-specific dyes and a second fluorescent dye selected from one of nucleic acid-specific dyes.

The blood detection method of claim 16, wherein the method further comprises: platelets and reticulocytes are distinguished based on fluorescence intensity and side scatter intensity information.

The blood detection method of any one of claims 1-17, wherein the method further comprises: the platelets are counted based on the optical information of the platelets obtained.

The blood detection method of any one of claims 1-18, wherein the optical detection system comprises: an optical subsystem, a flow cell, a first detector;

the optical subsystem includes: the laser comprises a laser, a front optical component and a rear optical component, wherein the front optical component comprises an optical isolator; wherein the content of the first and second substances,

the laser configured to emit a laser beam;

the front light assembly is configured to perform front light treatment on the laser beam, and the laser beam subjected to the front light treatment is converged at the blood cell measured sample of the flow chamber in a first direction and generates scattered light;

the rear light assembly is arranged behind the flow chamber along the propagation direction of the laser beam and is configured to perform rear light treatment on the scattered light, so that the scattered light subjected to the rear light treatment enters the first detector to be subjected to light intensity detection;

the optical isolator configured to isolate reflected light from the laser; the reflected light is generated by the laser beam passing through the flow chamber.

The method of claim 19, wherein the optical isolator comprises a beam splitter prism and a polarization conversion module that are adhesively connected to each other;

the beam splitting prism is configured to reflect an S-polarized component of the incident laser beam and transmit a P-polarized component of the incident laser beam;

the polarization state conversion assembly is configured to change the polarization state of the P polarization component transmitted by the light splitting prism, so that the P polarization component is changed from linearly polarized light to circularly polarized light, and the polarization state after the circularly polarized light is reflected is changed into S polarized light and is reflected by the light splitting prism.

The blood detection method of claim 19, wherein the optical isolator is composed of an analyzer and a polarization state conversion element that are adhesively connected to each other;

the analyzer is configured to allow only a P-polarized component of the laser beam to pass through;

the polarization state conversion component is configured to change the polarization state of the P polarization component passing through the analyzer, so that the P polarization component is changed from linearly polarized light to circularly polarized light, and the polarization state of reflected light of the circularly polarized light is changed into S polarized light and is isolated by the analyzer.

The method according to claim 19, wherein the optical isolator comprises a band-pass filter and a frequency doubling crystal which are connected with each other by adhesion;

the band-pass filter is configured to pass the laser beam with the wavelength lambda;

and the frequency doubling crystal is configured to double the frequency of the laser beam passing through the band-pass filter, and double the frequency of the reflected light of the laser beam after frequency doubling again, so that the reflected light is filtered by the band-pass filter.

The blood testing method of claim 19, wherein the front light assembly further comprises a collimating lens;

the collimating lens is arranged between the laser and the optical isolator along the propagation direction of the laser beam and is configured to collimate the laser beam so that the laser beam becomes a parallel beam.

The blood testing method of claim 19, wherein the backlight assembly further comprises a stop;

the front light assembly is further configured to perform front light processing on the laser beam, so that the front light processed laser beam is converged at the straight stop diaphragm in a second direction.

The blood detection method of claim 19, wherein the front light assembly further comprises a first light collection assembly and a second light collection assembly;

the first light converging component is configured to perform first focusing on the laser beam, so that the laser beam is converged at the blood cell measured sample of the flow chamber in a first direction and generates scattered light;

the second light converging component is configured to perform second focusing on the laser beam, so that the laser beam is converged at the straight stop included in the rear light component in a second direction.

The blood testing method of claim 19, wherein the rear light assembly further comprises a third converging assembly and an aperture stop;

the third converging component is configured to perform third focusing on the scattered light, so that the scattered light is converged at the aperture stop and enters the first detector through the aperture of the aperture stop.

A method of blood testing, the method comprising:

treating a blood sample with a first reagent to obtain a test sample, wherein the first reagent comprises a hemolytic agent which lyses red blood cells in the blood sample;

enabling particles in a sample to be detected to pass through a detection area of an optical detection system one by one, and acquiring optical information of the sample to be detected; and

optical information of the platelets is obtained from at least two of the optical information.

The method of claim 27, wherein the hemolytic agent lyses red blood cells in the blood sample into fragments having light scattering properties significantly different from platelets.

The blood detection method of claim 28, wherein the method counts platelets based on forward scattered light intensity and side scattered light intensity.

The blood detection method of claim 28 or 29, wherein the method further distinguishes leukocytes into subpopulations including at least monocytes, lymphocytes, and neutrophils based on forward scattered light intensity and side scattered light intensity.

The blood detection method of claim 27, the method further comprising: processing the blood sample with a second reagent, wherein the second reagent comprises a fluorescent dye selected from the group consisting of membrane-specific dyes and mitochondrial-specific dyes, or comprises a fluorescent dye selected from the group consisting of nucleic acid-specific dyes, to further obtain a fluorescent signal to distinguish between platelets and reticulocytes based on the scattered light intensity and fluorescence intensity information.

The blood detection method of claim 31, wherein the method further comprises: differentiating the subpopulation of leukocytes based on the fluorescence intensity and the side scattered light intensity information to obtain subpopulations of leukocytes including at least monocytes, lymphocytes and neutrophils and/or identify naive granulocytes.

The method for detecting blood according to claim 31, wherein the platelets are totally distinguished based on fluorescence intensity and forward scattered light intensity information.

The blood detection method of claim 31, wherein the method further comprises: the reticulocytes are alarmed when the number of particles in a preset area of a scatter diagram formed from the forward scattered light intensity and the fluorescence intensity exceeds a predetermined threshold.

The blood detection method of claim 27, wherein the method further comprises: processing the blood sample with a second reagent, wherein the second reagent comprises a fluorescent dye selected from the group consisting of membrane-specific dyes and mitochondrial-specific dyes, and a fluorescent dye selected from the group consisting of nucleic acid-specific dyes, obtaining a fluorescent signal, and distinguishing platelets from reticulocytes based on scattered light intensity and fluorescence intensity information.

The blood detection method of claim 35, wherein the method further comprises: platelets and reticulocytes are distinguished based on fluorescence intensity and forward scattered light intensity information, and preferably further counted.

The blood detection method of any one of claims 27, wherein the optical detection system comprises: an optical subsystem, a flow cell, a first detector;

the optical subsystem includes: the laser comprises a laser, a front optical component and a rear optical component, wherein the front optical component comprises an optical isolator; wherein the content of the first and second substances,

the laser configured to emit a laser beam;

the front light assembly is configured to perform front light treatment on the laser beam, and the laser beam subjected to the front light treatment is converged at the blood cell measured sample of the flow chamber in a first direction and generates scattered light;

the rear light assembly is arranged behind the flow chamber along the propagation direction of the laser beam and is configured to perform rear light treatment on the scattered light, so that the scattered light subjected to the rear light treatment enters the first detector to be subjected to light intensity detection;

the optical isolator configured to isolate reflected light from the laser; the reflected light is generated by the laser beam passing through the flow chamber.

The method of claim 37, wherein the optical isolator comprises a beam splitter prism and a polarization conversion module adhesively connected to each other;

the beam splitting prism is configured to reflect an S-polarized component of the incident laser beam and transmit a P-polarized component of the incident laser beam;

the polarization state conversion component is configured to change the polarization state of the P polarization component transmitted by the light splitting prism, so that the P polarization component is changed from linearly polarized light to circularly polarized light, and the polarization state after the circularly polarized light is reflected is changed into S polarized light and is reflected by the light splitting prism; or

The optical isolator consists of an analyzer and a polarization state conversion component which are connected with each other in a bonding mode;

the analyzer is configured to allow only a P-polarized component of the laser beam to pass through;

the polarization state conversion component is configured to change the polarization state of the P polarization component passing through the analyzer, so that the P polarization component is changed from linearly polarized light to circularly polarized light, and the polarization state of reflected light of the circularly polarized light is changed into S polarized light and is isolated by the analyzer.

The method according to claim 27, wherein the two optical information are forward scattered light intensity information and side scattered light intensity information, and the platelets are counted and/or the leukocytes are differentiated into at least monocyte, lymphocyte and neutrophil subpopulations according to the forward scattered light intensity information and the side scattered light intensity information.

A blood analysis system comprising:

a sampling part for obtaining a blood sample and transferring the blood sample to the reaction part;

a reagent supply section for storing at least a first reagent and supplying it to the reaction section as required;

a reaction part including a mixing chamber for mixing the blood sample with a first reagent to form a test sample, wherein the first reagent includes a hemolytic agent which lyses red blood cells in the blood sample;

the optical detection system comprises a flow chamber and at least a first detector, and the first detector is used for detecting particles in the sample to be detected to obtain optical information in the sample to be detected when the sample to be detected is conveyed to the optical system from the mixing chamber and the particles in the sample to be detected pass through the flow chamber one by one to reach a detection area; and

a data processing module operatively connected with the optical system and comprising a processor and a non-transitory computer readable storage medium storing a computer program, wherein the computer program when executed by the processor performs the steps of: optical information of the platelets is obtained from at least two of the optical information.

The blood analysis system of claim 40, wherein the hemolytic agent lyses red blood cells in the blood sample into debris having light scattering properties significantly different from platelets, and wherein in the processing module, when the computer program is executed by the processor, further performs the steps of:

counting platelets based on the intensity of forward scattered light and the intensity of side scattered light in the obtained optical information; and/or

The leukocytes are differentiated into at least monocyte, lymphocyte and neutrophil subpopulations based on the intensity of the forward scattered light and the intensity of the side scattered light.

The blood analysis system of claim 40,

the mixing chamber is used for mixing the blood sample with a second reagent to form a test sample, wherein the second reagent comprises a fluorescent dye selected from a membrane-specific dye and a mitochondria-specific dye;

the optical detection system comprises a second detector which is a fluorescence detector so as to further obtain fluorescence signals when particles in the sample to be detected pass through a detection area one by one; and

in the data processing module, when the computer program is executed by the processor, the following steps are further performed:

differentiating the subpopulation of leukocytes according to the fluorescence intensity and the side scattered light intensity information to obtain subpopulations of leukocytes including at least monocytes, lymphocytes and neutrophils and/or identify naive granulocytes;

fully distinguishing the platelets according to the fluorescence intensity and the forward scattered light intensity information; and/or

The reticulocytes are alarmed when the number of particles in a preset area of a scatter diagram formed according to the forward scattered light intensity and fluorescence intensity information exceeds a predetermined threshold value.

The blood analysis system of claim 42, wherein the second reagent further comprises a fluorescent dye selected from nucleic acid specific dyes, and when the computer program is executed by the processor further performs the steps of: platelets and reticulocytes are distinguished based on fluorescence intensity and scattered light intensity information, and preferably further counted.

The blood analysis system of claim 40, wherein the optical detection system further comprises: an optical sub-system is provided which comprises,

the optical subsystem includes: the laser comprises a laser, a front optical component and a rear optical component, wherein the front optical component comprises an optical isolator; wherein the content of the first and second substances,

the laser configured to emit a laser beam;

the front light assembly is configured to perform front light treatment on the laser beam, and the laser beam subjected to the front light treatment is converged at the blood cell measured sample of the flow chamber in a first direction and generates scattered light;

the rear light assembly is arranged behind the flow chamber along the propagation direction of the laser beam and is configured to perform rear light treatment on the scattered light, so that the scattered light subjected to the rear light treatment enters the first detector to be subjected to light intensity detection;

the optical isolator configured to isolate reflected light from the laser; the reflected light is generated by the laser beam passing through the flow chamber.