CN108303363B - Blood cell analysis method and blood cell analyzer using same - Google Patents
Blood cell analysis method and blood cell analyzer using same Download PDFInfo
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- CN108303363B CN108303363B CN201710021197.0A CN201710021197A CN108303363B CN 108303363 B CN108303363 B CN 108303363B CN 201710021197 A CN201710021197 A CN 201710021197A CN 108303363 B CN108303363 B CN 108303363B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/1031—Investigating individual particles by measuring electrical or magnetic effects thereof, e.g. conductivity or capacity
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Electro-optical investigation, e.g. flow cytometers
- G01N15/1434—Electro-optical investigation, e.g. flow cytometers using an analyser being characterised by its optical arrangement
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N2015/1006—Investigating individual particles for cytology
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Electro-optical investigation, e.g. flow cytometers
- G01N2015/1477—Multiparameters
Abstract
The invention discloses a blood cell analysis method and a blood cell analyzer using the same. The method utilizes a more reasonable flow, so that a reliable five-class white blood cell result can be obtained in one reaction tank by utilizing a time-sharing multiplexing technology, and meanwhile, the measurement time is saved, the instrument complexity is simplified, and the cost is reduced.
Description
Technical Field
The present invention relates to a blood cell analysis method and a blood cell analyzer.
Background
The blood cell analyzer is mainly used for detecting the content of each blood cell in the peripheral blood of a human body and mainly comprises the total number of red blood cells, the total number of platelets, the total number of white blood cells, five sub-populations of white blood cells and the like. Wherein the leukocyte subpopulations are lymphocytes, monocytes, neutrophils, basophils and eosinophils, respectively. In addition, the blood cell analyzer can also detect the hemoglobin concentration (HGB) of red blood cells in blood. And derived parameters based on the above parameters.
Most of the blood cell analyzers in the current market adopt a two-channel scheme to realize five classification of white blood cells: two reaction tanks are adopted to respectively carry out a leukocyte tetra-classification (DIFF) reaction and a BASO reaction, and then the two reaction tanks are respectively sent to the same photoelectric detection unit or respective independent corresponding detection units. In the DIFF channel, the reagent will lyse erythrocytes in the blood and process leukocytes in the blood, such that the four classes of leukocytes (lymphocytes, monocytes, neutrophils and eosinophils) aggregate and classify in both dimensions of size and internal complexity. After the sample to be tested of the DIFF channel is sent to the laser scattering detection unit, the unit detects the information of the two dimensions, thereby four-classifying the white blood cells. In the BASO channel, the reagent will dissolve erythrocytes in blood and shrink all other cells except basophils in leukocytes, and the count value of basophils can be obtained after the cells are sent to a detection unit (the detection unit may be a laser scattering detection unit as shown in fig. 1, or may be an electrical impedance detection unit as shown in fig. 2). After the results of the two channels are combined, complete five-class information of the white blood cells can be calculated and obtained.
In such a blood cell analyzer, a separate red blood cell detection channel is generally provided, and the count values of red blood cells and platelets are detected by an electrical impedance method, and the hemoglobin content in blood is generally detected in a BASO channel, or a separate dedicated channel. The checking flow is shown in fig. 1 and 2.
For differential detection of leukocytes, two separate detection channels are required. The method has the advantages that five kinds of white blood cells can be accurately and reliably classified, and the detection result of HGB can be obtained in the BASO detection channel. But the disadvantages are more obvious: two reactions are needed, the blood consumption is high, and the reagent types are various; two reaction tanks and two sets of sample liquid conveying pipelines are needed, the volume of the instrument is large, the structure is complex, and the cost is high.
Disclosure of Invention
The invention aims to provide a blood cell analysis method and a blood cell analyzer using the same, which not only can obtain reliable five-class results of white blood cells, but also can reduce the dosage of blood and reagents, simplify the structure of an instrument, reduce the volume of the instrument and reduce the cost of the instrument.
According to a first aspect of the present invention, there is provided a blood cell analysis method comprising treating a reaction cell of a blood sample with reagents, filling various reagents and a liquid path system of the sample, and a laser light scattering detection system for detecting blood cells in the treated sample, the flow of the analysis method being as follows:
injecting a blood sample and a first group of reagents into a reaction tank, wherein the first group of reagents are used for diluting the blood sample, and diluting to obtain a diluted sample liquid;
pumping a proper amount of diluted sample liquid out of the reaction tank and temporarily storing the diluted sample liquid;
injecting a second group of reagents into the reaction tank, and reacting with the rest sample diluent in the reaction tank to obtain a second sample liquid;
delivering a proper amount of second sample liquid to the laser scattering detection system for classifying and detecting the white blood cells;
evacuating the residual second sample liquid in the reaction tank, and cleaning a relevant liquid path and a laser scattering detection system through which the second sample liquid passes in the conveying process, but not cleaning the reaction tank;
and injecting a proper amount of the temporarily stored diluted sample liquid and a third group of reagents into a reaction tank, and reacting to obtain a third sample liquid.
The reaction tank is only one, the second sample liquid is used for four-class detection of leucocytes, a proper amount of the third sample liquid is conveyed to the laser scattering detection system for class detection of basophils, and the third sample liquid is also used for hemoglobin detection.
The method uses laser scattering detection data of the second sample liquid and/or the third sample liquid to count the total number of white blood cells.
The four actions of detecting the second sample liquid in the laser scattering detection system, evacuating the second sample liquid in the reaction tank, cleaning the second sample liquid flowing through the pipeline and the laser scattering detection system, filling the third group of reagents and diluting the sample liquid into the reaction tank can be partially performed simultaneously or in a time-sharing manner.
The second sample fluid may also be used directly for five-class detection of leukocytes, in which case the third sample fluid is used only for hemoglobin detection.
According to a second aspect of the present invention, there is provided a blood cell analyzer comprising:
only one reaction cell for mixing a blood sample with a reagent to prepare a sample liquid; only one laser scattering detection device for detecting white blood cells; the device is provided with a corresponding liquid path device for filling and conveying sample liquid and/or reagent; the device is provided with a corresponding control device which is combined with a liquid path device, a laser scattering detection device and the like and is used for preparing a sample liquid, detecting the sample liquid and cleaning and evacuating pipelines according to the modes of claims 1-9.
The liquid path device comprises a liquid-gas path power source such as an injection pump, a positive and negative air pressure source, a DP pump and the like, related controllable valves, pipelines and the like.
The laser scattering detector includes at least a semiconductor laser, an aspherical lens for laser shaping, and the like.
Drawings
FIG. 1 is a flow chart of a prior art blood cell analysis method;
FIG. 2 is a flow chart of another prior art blood cell analysis method;
FIG. 3 is a flow chart of a single channel blood cell analysis method 1;
FIG. 4 is a flow chart of a single channel blood cell analysis method 2;
fig. 5 is a flow chart of a blood analysis method provided by the invention.
Detailed Description
The invention is described in further detail below with reference to the drawings and examples.
In order to overcome the defects of complex structure, large volume and high cost of a double-channel scheme instrument, the scheme which is easiest to think is a single-channel method. The method comprises the steps of adopting a leukocyte reaction tank, utilizing more optimized reagent components, carrying out five classifications on leukocytes after the obtained sample liquid passes through a laser scattering detection system only through one reaction, and then detecting the HGB content in blood in the same reaction tank by utilizing a time-sharing multiplexing technology. The flow is shown in fig. 3.
Or a leukocyte reaction tank is adopted, a DIFF reagent is added first, laser scattering detection is carried out after the reaction is finished, the pipeline is emptied and cleaned after the detection is finished, and then a BASO reagent and a corresponding sample are added for HGB detection and laser scattering detection. The flow is shown in fig. 4.
The two schemes use time division multiplexing technology to serially carry out two groups of different reagent reactions, and have the advantages that one set of liquid path system and one set of laser scattering detection system are needed, and the disadvantage that the serial operation is needed and the single test time of the instrument is longer. More importantly, for the first protocol, due to the specificity of basophils, it is difficult to classify them from other leukocytes in a single reaction, and it is difficult to distinguish them thoroughly, which results in inaccurate measurement results.
The analysis method provided by the invention is an optimization of the second scheme of the time division multiplexing single channel method on the basis of researching the characteristics of the DIFF reagent and the BASO reagent.
For the above blood cell analysis method, the main function of the DIFF reagent is to lyse red blood cells in a blood sample and to process white blood cells in the blood sample so that the four classes of white blood cells (lymphocytes, monocytes, neutrophils and eosinophils) appear to be aggregated differently in both dimensions of size and internal complexity and to be classified into distinct four classes, and in fact to produce a distinct shrinkage effect on all white blood cells. The BASO reagent has the main function of obviously shrinking all the white blood cells except the BASO granulocytes by utilizing the characteristics of the BASO granulocytes, so that the volume of the white blood cells is reduced. Therefore, in principle, even if a small amount of DIFF reagent remains in the sample when the BASO reagent acts, the detection result is not affected.
Based on the above principle, the present invention proposes a new optimized blood cell analysis method, the flow chart of which is shown in fig. 5, and the method will be described in detail below with reference to the drawings.
The first step is to suck the sample, then add the sample into the leukocyte reaction cell, and add the first group of reagents, which have the main function of diluting the blood sample so that the sample liquid after the subsequent treatment has a proper dilution ratio. The diluted sample liquid after the treatment is divided into three parts, wherein one part of the diluted sample liquid is reserved in a leucocyte reaction tank, one part of the diluted sample liquid is injected into a red blood cell reaction tank, and the other part of the diluted sample liquid is temporarily stored in a sample liquid suction pipeline.
In the leucocyte reaction cell, a diluted sample liquid with a fixed volume is reserved, and a second group of reagents, which can be one or more reagents, is added into the leucocyte reaction cell. And the second group of reagents are fully and uniformly mixed with the diluted sample liquid, and after corresponding chemical reaction, a second sample liquid to be detected is formed. And conveying a proper amount of second sample liquid to a laser scattering detection unit for detection, and performing DIFF four-classification on the white blood cells after measurement. After the second sample liquid is conveyed, the leucocyte reaction tank is emptied, and the action can be performed simultaneously or in a time-sharing manner with the action of detecting the second sample liquid by the laser scattering unit.
After the blank cell reaction tank is discharged, according to the principle, the diluted sample temporarily stored in the sample liquid suction pipeline can be injected into the emptied cell reaction tank without cleaning the reaction tank, and meanwhile or in a time-sharing manner, a third group of reagents can be one or more reagents. And after the third group of reagents and the diluted sample liquid are fully and uniformly mixed and undergo corresponding chemical reactions, forming a third sample liquid in the leucocyte reaction cell.
After the detection of the second sample liquid is completed, the conveying pipeline of the second sample liquid and the laser scattering detection unit are cleaned (the unit can be cleaned) so as to avoid cross contamination, and the cleaning pipeline action and the preparation action of the third sample liquid can be performed simultaneously or in a time-sharing manner.
After the preparation of the third sample liquid is finished, the second sample liquid conveying pipeline is cleaned, a proper amount of the third sample liquid is conveyed to the laser scattering detection unit, if the laser scattering detection unit is not cleaned before, the detection unit is cleaned by using a reagent in the pipeline and a conveyed power source while conveying, and if the laser scattering detection unit is cleaned before, the third sample liquid is directly conveyed to the laser scattering detection unit.
After the third sample liquid enters a laser scattering detection unit, the unit detects the content of basophils in the white blood cells, and the five classification results of the white blood cells in the blood can be obtained by combining the detection result of the DIFF obtained before.
In the preparation process of the third sample liquid in the leukocyte reaction tank, or after the preparation is finished, and the HGB detection device arranged on the leukocyte reaction tank can be started to detect the HGB content in blood, the action can be performed simultaneously with or in a time-sharing way with the action of conveying the third sample liquid to the laser scattering detection unit.
The relevant flow actions in the red blood cell reaction cell are completely independent of the reactions or actions in the white blood cell reaction cell. After the diluted sample solution is injected into the erythrocyte reaction cell, the erythrocyte reagent is injected simultaneously or in a time-sharing manner, and generally, the reagent mainly aims at diluting blood. After the mixture is fully and evenly mixed and reacts, the preparation of the erythrocyte sample liquid in the erythrocyte reaction tank is completed, and the sample liquid has a proper dilution ratio. Then, an electrical impedance method detection unit arranged on the erythrocyte reaction tank is started, and meanwhile, relevant liquid and gas paths are matched, so that the detection of the erythrocyte sample liquid is completed. The measurement will yield parameters related to the red blood cells and platelets in the blood.
The invention also provides a blood cell analyzer using the blood cell analysis method: only one leukocyte reaction cell was used to detect five classes of leukocytes and HGB; only one laser scattering detection unit is used for detecting the internal complexity of the size of the cells in the sample liquid, namely the scattering condition of the cells on laser; only one erythrocyte reaction cell detects the content of erythrocytes and platelets;
which contains associated fluid path systems and control systems to enable the overall sample and reagent processing flow, and the workflow of each detection unit to operate in the manner provided by the present invention as described above.
The foregoing is a further detailed description of embodiments of the invention and is not intended to limit the invention to the precise form described. Modifications which can be easily obtained by a person skilled in the art by simple changes or substitutions, without departing from the spirit of the invention, shall also be considered as falling within the scope of the invention.
Claims (10)
1. A blood cell analysis method comprising a reaction cell for treating a blood sample with a reagent, a liquid path system for filling various reagents and samples, and a laser light scattering detection system for detecting blood cells in the treated sample, characterized in that:
injecting a blood sample and a first group of reagents into a reaction tank, wherein the first group of reagents are used for diluting the blood sample, and diluting to obtain a diluted sample liquid; the diluted sample liquid is divided into three parts, wherein one part of the diluted sample liquid is reserved in a leucocyte reaction tank, one part of the diluted sample liquid is injected into a erythrocyte reaction tank, and the other part of the diluted sample liquid is temporarily stored in a sample liquid suction pipeline;
pumping a proper amount of diluted sample liquid out of the reaction tank and temporarily storing the diluted sample liquid;
injecting a second group of reagents into the reaction tank, and reacting with the rest sample diluent in the reaction tank to obtain a second sample liquid; the second sample liquid is used for four-classification detection of white blood cells;
delivering a proper amount of second sample liquid to the laser scattering detection system for classifying and detecting the white blood cells;
evacuating the residual second sample liquid in the reaction tank, and cleaning a relevant liquid path and a laser scattering detection system through which the second sample liquid passes in the conveying process, but not cleaning the reaction tank;
injecting a proper amount of temporarily stored diluted sample liquid and a third group of reagents into a reaction tank, and reacting to obtain a third sample liquid; the third sample fluid is delivered to the laser light scattering detection system for differential detection of basophils.
2. The blood cell analysis method according to claim 1, wherein: only one reaction tank is provided.
3. The blood cell analysis method according to claim 1 or 2, wherein: the third sample liquid is also used for detecting hemoglobin.
4. The blood cell analysis method according to claim 1 or 2, wherein: and counting the total white blood cells by using the laser scattering detection data of the second sample liquid and/or the third sample liquid.
5. The blood cell analysis method according to claim 1, wherein: the second sample liquid enters a laser scattering detection system for detection, the second sample liquid in the reaction tank is emptied, the second sample liquid is cleaned, flows through a pipeline and the laser scattering detection system, and a third group of reagents and diluted sample liquid are filled into four action parts of the reaction tank and are performed simultaneously or in a time-sharing manner.
6. The blood cell analysis method according to claim 1 or 2, wherein: the second sample liquid is used for five-class detection of the white blood cells.
7. The method for analyzing blood cells according to claim 1, 2 or 4, wherein: the third sample liquid is used for detecting hemoglobin.
8. A blood cell analyzer, comprising:
only one reaction cell for mixing a blood sample with a reagent to prepare a sample liquid;
only one laser scattering detection device for detecting white blood cells;
the liquid path device is used for filling and conveying sample liquid and/or reagent;
control means, which are combined with the liquid path means, the laser scattering detection means, for preparing the sample liquid, detecting the sample liquid and cleaning and evacuating the pipe according to the method of any one of claims 1 to 7.
9. The blood cell analyzer of claim 8, wherein: the liquid path device comprises a plurality of injection pumps, positive and negative air pressure sources, a liquid-air path power source of the DP pump, related pipelines and controllable valves.
10. The blood cell analyzer of claim 8 or 9, wherein: the laser scattering detection device at least comprises a semiconductor laser and an aspheric lens for laser shaping.
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| CN111239027A (en) * | 2020-01-22 | 2020-06-05 | 深圳市锦瑞生物科技有限公司 | Blood particle detection method and blood analyzer |
| CN112481346B (en) * | 2020-12-03 | 2022-12-09 | 中国人民解放军陆军军医大学第二附属医院 | Automatic early warning system and method for detecting abnormal cells in peripheral blood cell morphology |
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