CN111381017A

CN111381017A - Five-classification hemolytic agent for white blood cells, preparation method, blood analyzer and detection method

Info

Publication number: CN111381017A
Application number: CN201811613080.2A
Authority: CN
Inventors: 郑剑通; 其他发明人请求不公开姓名
Original assignee: Shenzhen Dymind Biotechnology Co Ltd
Current assignee: Shenzhen Dymind Biotechnology Co Ltd
Priority date: 2018-12-27
Filing date: 2018-12-27
Publication date: 2020-07-07
Anticipated expiration: 2038-12-27
Also published as: CN111381017B

Abstract

The application discloses five-classification hemolytic agent for white blood cells, a preparation method, a blood analyzer and a detection method, wherein the five-classification hemolytic agent for white blood cells comprises the following components: surfactants, buffers and cosolvents; wherein the surfactant comprises an amino acid type surfactant and at least one of a quaternary ammonium salt cationic surfactant or a polyethylene glycol type nonionic surfactant. Through the mode, the foaming of the hemolytic agent can be reduced, interference factors in the detection process are further reduced, the use cost is reduced, and the detection steps are reduced.

Description

Five-classification hemolytic agent for white blood cells, preparation method, blood analyzer and detection method

Technical Field

The application relates to the technical field of blood cell analysis, in particular to a five-classification hemolytic agent for white blood cells, a preparation method, a blood analyzer and a detection method.

Background

In clinical hematology tests, the counting and classification of peripheral blood leukocytes in patients are of great importance for the diagnosis and treatment of many diseases. In recent years, hemolytic agents generally consist of hemolytic agent a and hemolytic agent B, wherein hemolytic agent a is used for dissolving red blood cells, and hemolytic agent B plays a stabilizing role.

In a long-term research and development process, the inventor of the present application finds that when the hemolytic agent is actually applied and a blood analyzer runs, the hemolytic agent B needs to be added after the hemolytic agent a is added for a certain time, which not only increases the use cost, but also increases the detection steps, thereby causing a problem of low detection efficiency. Meanwhile, most of the existing hemolytic agents contain carbonate ions and bicarbonate ions, so that bubbles are easy to generate, interference factors are more in the detection process, particle counting is affected, and the problems of inaccurate test signals and large errors are solved.

Disclosure of Invention

The technical problem mainly solved by the application is to provide a five-classification leukocyte hemolytic agent, a preparation method, a blood analyzer and a detection method, and the problems that interference factors are more, errors are large and the detection efficiency is low in the detection process can be solved.

In order to solve the technical problem, the application adopts a technical scheme that: provided is a hemolyzing agent for five-classification of white blood cells, which comprises: surfactants, buffers and cosolvents; wherein the surfactant comprises an amino acid type surfactant and at least one of a quaternary ammonium salt cationic surfactant or a polyethylene glycol type nonionic surfactant.

Wherein, the five-classification hemolytic agent for the white blood cells further comprises: and (4) a preservative.

Wherein, the concentration of the surface active agent is 0.1 to 7.0g/L, the concentration of the buffering agent is 2.0 to 10.0g/L, the concentration of the cosolvent is 3.0 to 15.0g/L, and the concentration of the preservative is 0.1 to 3.0 parts g/L.

Wherein the amino acid type surfactant comprises at least one of sodium lauroyl glutamate, sodium lauroyl sarcosinate, sodium cocoyl glycinate, cocoyl triethanolamine glutamate, sodium palmitoyl glutamate, cocoyl propyl glutamate, disodium cocoyl glutamate, sodium cocoyl glutamate, N-myristyl-L-glutamic acid dibutylamide, or sodium octadecyl dimethyleneaminodicarboxylate; the quaternary ammonium salt cationic surfactant comprises at least one of cocamidopropyl dimethyl amine lactate, hydroxyethyl lauryl dimethyl ammonium chloride, dodecyl dimethyl benzyl ammonium chloride, tetradecyl trimethyl ammonium chloride, hexadecyl trimethyl ammonium chloride, dodecyl trimethyl ammonium chloride, hexadecyl trimethyl ammonium bromide, dodecyl trimethyl ammonium bromide or tetradecyl trimethyl ammonium bromide; the polyethylene glycol nonionic surfactant comprises at least one of lauryl polyoxyethylene (7) ether, polyethylene glycol p-isooctyl phenyl ether, polyoxyethylene fatty amide or stearic acid amide polyoxyethylene (6) ether.

Wherein the buffer comprises at least one of acetic acid/potassium acetate buffer solution, tris/hydrochloric acid buffer solution, 3- (N-morpholinyl) propanesulfonic acid sodium salt/hydrochloric acid buffer solution, or disodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt; the cosolvent comprises at least one of ethylene glycol, methanol, ethanol, isopropanol or glycerol.

Wherein the antiseptic is at least one of Kathon, potassium sorbate or benzoic acid.

In order to solve the technical problem, the application adopts a technical scheme that: provides the application of the five-classification hemolytic agent for the white blood cells in the five-classification counting of the white blood cells.

In order to solve the technical problem, the application adopts a technical scheme that: a preparation method of a leukocyte five-classification hemolytic agent is provided, and the preparation method comprises the following steps: weighing a surfactant, a buffering agent, a cosolvent and a preservative according to a preset formula, wherein the surfactant comprises an amino acid type surfactant and at least one of a quaternary ammonium salt cationic surfactant or a polyethylene glycol type nonionic surfactant; pouring a surfactant, a buffering agent, a cosolvent and a preservative into a dosing barrel filled with water; stirring to completely dissolve the surfactant, the buffer, the cosolvent and the preservative; filtering to obtain leukocyte five-classification hemolytic agent; wherein, in the leukocyte five-classification hemolytic agent, the concentration of the surfactant is 0.1-7.0g/L, the concentration of the buffer is 2.0-10.0g/L, the concentration of the cosolvent is 3.0-15.0g/L, and the concentration of the preservative is 0.1-3.0 g/L.

In order to solve the technical problem, the application adopts a technical scheme that: provided is a blood analyzer including: a hemolytic agent bottle and a hemolytic agent injector communicated with the hemolytic agent bottle; the hemolytic agent bottle is used for containing the leukocyte five-classification hemolytic agent, and the hemolytic agent injector is used for sucking the hemolytic agent from the hemolytic agent bottle and pushing the leukocyte five-classification hemolytic agent into the reaction tank so as to mix the leukocyte five-classification hemolytic agent with the blood sample to be analyzed.

In order to solve the technical problem, the application adopts a technical scheme that: the detection method based on the blood analyzer comprises the following steps: sucking a blood sample to be detected into a reaction pool; sucking the leukocyte five-classification hemolytic agent from the hemolytic agent bottle through a hemolytic agent injector, and pushing the leukocyte five-classification hemolytic agent into the reaction tank; mixing a leukocyte five-classification hemolytic agent with a blood sample to be detected; and detecting the size and morphological information of the white blood cells through a white blood cell channel measuring module.

The beneficial effect of this application is: different from the prior art, the leukocyte five-classification hemolytic agent can be used for determining parameters such as total number of leukocytes, leukocyte five-classification counting and the like after being mixed with a blood sample. Because the amino acid type surfactant selected by the application is used as the surfactant and is matched with a proper amount of quaternary ammonium salt cationic surfactant or polyethylene glycol nonionic surfactant, the rapid dissolution of erythrocytes can be realized through the surfactant, the foaming of the hemolytic agent can be reduced by using the amino acid type surfactant, and then interference factors in the detection process are reduced, and the buffer and the cosolvent added in the application can ensure that a diluent system has a stable pH value environment, therefore, when a blood analyzer operates, only the leukocyte five-classification hemolytic agent in the application needs to be added, the hemolytic agent B or hemolytic agent C does not need to be additionally added, the erythrocytes can be dissolved and the leukocytes can be stabilized, the use cost is reduced, and the detection steps are reduced. Meanwhile, the leukocyte five-classification hemolytic agent does not contain carbonate ions, bicarbonate ions and anionic surfactants, so that foaming is not easy to occur, interference factors in the detection process are reduced, accurate leukocyte five-classification counting is obtained, the correlation of the determination result is good, and the accuracy is high.

Drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings needed to be used in the description of the embodiments are briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present application, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without creative efforts. Wherein:

FIG. 1 is a schematic flow chart of one embodiment of a process for preparing a five-classification hemolytic agent for leukocyte of the present application;

fig. 2 is a schematic flow chart of an embodiment of the detection method based on the blood analyzer.

Detailed Description

The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application, and it is obvious that the described embodiments are only a part of the embodiments of the present application, and not all the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.

The application provides a five-classification hemolytic agent for white blood cells, which comprises the following components: surfactants, buffers, and cosolvents. Wherein the surfactant comprises an amino acid type surfactant and at least one of a quaternary ammonium salt cationic surfactant or a polyethylene glycol type nonionic surfactant.

Specifically, five-classification hemolytic agents for leukocytes can divide leukocytes into five cell populations, which are neutrophils, lymphocytes, monocytes, eosinophils, and basophils, respectively. The five-classification hemolytic agent for white blood cells contains a surfactant, and because red blood cells are anucleate, contain more cholesterol than white blood cells, and have easily broken and deformed cell membranes, the five-classification hemolytic agent for white blood cells can penetrate into red blood cell cytoplasm more quickly to cause swelling and rupture of red blood cells, so that the red blood cells are dissolved before the white blood cells.

The amino acid type surfactant has a certain immobilization effect on cytoplasm and cell membrane of leucocyte while dissolving erythrocyte. The polyethylene glycol nonionic surfactant, especially the polyethylene glycol nonionic surfactant with the molecular weight of 200-1500 can efficiently and quickly hemolysis and ensure the smooth and quick proceeding of the subsequent five-classification counting of the leucocytes. The quaternary ammonium salt cationic surfactant has the capacity of cracking erythrocytes, but has certain damage to leukocytes when the concentration of the quaternary ammonium salt cationic surfactant is more than or equal to 10 g/L.

Preferably, the surfactant in this embodiment may be a combination of an amino acid type surfactant and a polyethylene glycol type nonionic surfactant, instead of the quaternary ammonium salt cationic surfactant, because the quaternary ammonium salt cationic surfactant may damage the structure of the leukocytes, so that the leukocytes cannot be restored to a state close to the original state during the detection by the instrument, and the result of classifying the leukocytes into five categories is also affected. In other embodiments, the surfactant may also be an amino acid type surfactant and a quaternary ammonium salt cationic surfactant in combination, but the amount of the quaternary ammonium salt cationic surfactant needs to be strictly controlled (for example, the concentration of the quaternary ammonium salt cationic surfactant may be 0.1-5.0g/L, such as 0.1g/L, 1.0g/L, or 5.0g/L) to avoid structural damage to leukocytes as much as possible.

The cosolvent can dissolve cell debris generated by lysing erythrocytes, and enhance the solubility of the surfactant in the hemolytic agent, thereby preventing crystallization or precipitation in the agent. The cosolvent can be alcohol substances, the alcohol substances have the effect of helping dissolution, can promote the dissolution of key substances, and has the effect of accelerating the reaction process. The concentration of the alcohol is usually 3.0 to 15.0g/L, preferably 2 to 8 g/L.

In order to maintain the lysing ability of the quaternary ammonium salt in the hemolytic agent to erythrocytes, the pH of the hemolytic agent must be maintained in a suitable range. In order to stabilize the pH of the hemolytic agent in a suitable range, the hemolytic agent component described in this embodiment includes a buffer, and the effective buffering range of the buffer used in this embodiment may be between pH 3.5-9.2.

Different from the prior art, the hemolyzing agent for five-classification of white blood cells of the present embodiment can be used to determine the total number of white blood cells, the five-classification count of white blood cells, and other parameters after being mixed with a blood sample. Because the amino acid type surfactant selected by the embodiment is used as the surfactant and is matched with a proper amount of quaternary ammonium salt cationic surfactant or polyethylene glycol nonionic surfactant, rapid dissolution of erythrocytes can be realized through the surfactant, and the foaming of the hemolytic agent can be reduced by using the amino acid type surfactant, so that interference factors in the detection process are reduced, and the buffer and the cosolvent added in the application can ensure that a diluent system has a stable pH value environment. Meanwhile, the leukocyte five-classification hemolytic agent does not contain carbonate ions, bicarbonate ions and anionic surfactants, so that foaming is not easy to occur, interference factors in the detection process are reduced, accurate leukocyte five-classification counting is obtained, the correlation of the determination result is good, and the accuracy is high.

In one embodiment, the five leukocyte classification hemolytic agent further comprises: and (4) a preservative.

Specifically, the hemolytic agent for leukocyte five classification contains a preservative. The preservative can inhibit the breeding of microorganisms in the long-term storage process of the reagent, prevent the reagent from deteriorating, prolong the storage and use time of the reagent and ensure the stable performance of the reagent in the period of validity. The preservative which has no influence on the performance of the reagent can be selected, and common preservatives comprise benzoic acid and salts thereof, sorbic acid and salts thereof, hydantoin and isothiazolinone compounds (Proclin300), phenoxyethanol and formaldehyde solution which have certain preservative efficacy. The dosage of the preservative is 0.1-3.0g/L, and different concentrations of different preservatives can be selected to ensure that the function of the reagent is normal in the effective period. In the application, after the preservative is added into the five-classification hemolytic agent for leucocytes, the effective period of the five-classification hemolytic agent for leucocytes can be prolonged.

In one embodiment, the concentration of the surfactant is 0.1-7.0g/L, the concentration of the buffer is 2.0-10.0g/L, the concentration of the cosolvent is 3.0-15.0g/L, and the concentration of the preservative is 0.1-3.0 g/L.

Specifically, in this embodiment, the mass-volume concentration is 1 ‰ 1g/L, which is the mass number (g) of solute/volume (L) of the solution, and water may be used as the solvent for the hemolytic agent. The concentration of surfactant can be 0.1-7.0g/L (e.g., 0.1g/L, 1.0g/L, 3.0g/L, or 7.0g/L), the concentration of buffer can be 2.0-10.0g/L (e.g., 2.0g/L, 4.0g/L, 8.0g/L, or 10.0g/L), the concentration of co-solvent can be 3.0-15.0g/L (e.g., 3.0g/L, 5.0g/L, 10.0g/L, or 15.0g/L), and the concentration of preservative can be 0.1-3.0 parts g/L (e.g., 0.1g/L, 0.5g/L, 1.0g/L, or 3.0 g/L).

The surfactant can be a combination of an amino acid type surfactant and a polyethylene glycol type nonionic surfactant, wherein the concentration of the amino acid type surfactant can be 0.1-1.0g/L (such as 0.1g/L, 0.5g/L or 1.0g/L), and the concentration of the polyethylene glycol type nonionic surfactant can be 0.3-5.0g/L (such as 0.3g/L, 1.0g/L or 5.0 g/L).

The surfactant may also be a combination of an amino acid type surfactant and a quaternary ammonium salt cationic surfactant, wherein the concentration of the amino acid type surfactant may be 0.1-1.0g/L (e.g., 0.1g/L, 0.5g/L, or 1.0g/L) and the concentration of the quaternary ammonium salt cationic surfactant may be 0.1-5.0g/L (e.g., 0.1g/L, 0.5g/L, or 1.0 g/L).

The surfactant may also be a combination of an amino acid type surfactant, a quaternary ammonium salt cationic surfactant, and a polyethylene glycol type nonionic surfactant, wherein the concentration of the amino acid type surfactant may be 0.1 to 1.0g/L (e.g., 0.1g/L, 0.5g/L, or 1.0g/L), the concentration of the quaternary ammonium salt cationic surfactant may be 0.1 to 5.0g/L (e.g., 0.1g/L, 0.5g/L, or 1.0g/L), and the concentration of the polyethylene glycol type nonionic surfactant may be 0.3 to 5.0g/L (e.g., 0.3g/L, 1.0g/L, or 5.0 g/L).

In one embodiment, the amino acid type surfactant may include at least one of sodium lauroyl glutamate, sodium lauroyl sarcosinate, sodium cocoyl glycinate, cocoyl triethanolamine glutamate, sodium palmitoyl glutamate, cocoyl propyl glutamate, disodium cocoyl glutamate, sodium cocoyl glutamate, N-myristyl-L-glutamic acid dibutylamide, or sodium octadecyl dimethyleneamino diformate. The quaternary ammonium salt cationic surfactant may include at least one of cocamidopropyl dimethyl amine lactate, hydroxyethyl lauryl dimethyl ammonium chloride, dodecyl dimethyl benzyl ammonium chloride, tetradecyl trimethyl ammonium chloride, hexadecyl trimethyl ammonium chloride, dodecyl trimethyl ammonium chloride, hexadecyl trimethyl ammonium bromide, dodecyl trimethyl ammonium bromide, or tetradecyl trimethyl ammonium bromide. The polyethylene glycol-based nonionic surfactant may include at least one of laureth-7, p-isooctylphenyl ether, polyoxyethylene fatty amide, or polyoxyethylene-6 ether octadecanoic acid amide.

In one embodiment, the buffer comprises at least one of acetic acid and potassium acetate buffer, tris and hydrochloric acid buffer, 3- (N-morpholino) propanesulfonic acid sodium salt and hydrochloric acid buffer, or disodium hydrogen phosphate and sodium dihydrogen phosphate buffer salt. The cosolvent comprises at least one of ethylene glycol, methanol, ethanol, isopropanol or glycerol.

Specifically, in order to maintain the lysing ability of the surfactant in the hemolytic agent to erythrocytes, it is necessary that the pH value of the hemolytic agent is maintained in an appropriate range. In this embodiment, the suitable pH range of the hemolytic agent may be at least one of acetic acid and potassium acetate buffer, tris and hydrochloric acid buffer, 3- (N-morpholino) propanesulfonic acid sodium salt and hydrochloric acid buffer, or disodium hydrogen phosphate and sodium dihydrogen phosphate buffer salt for different surfactants. The buffer may be used at a concentration of 2.0 to 10.0 g/L.

Unlike the prior art that a nonionic surfactant or a quaternary ammonium cationic surfactant is used as a solubilizer, in this embodiment, at least one solvent of ethylene glycol, methanol, ethanol, isopropanol or glycerol is used as a cosolvent, so that damage of the nonionic surfactant or the quaternary ammonium cationic surfactant to leukocytes can be reduced.

In one embodiment, the preservative is at least one of kathon, potassium sorbate, or benzoic acid.

The application also provides application of the leukocyte five-classification hemolytic agent in blood leukocyte five-classification counting.

The application provides a preparation method of a leukocyte penta-classification hemolytic agent, which comprises the following steps:

s101: and weighing the surfactant, the buffer, the cosolvent and the preservative according to a preset formula.

Wherein the surfactant comprises an amino acid type surfactant and at least one of a quaternary ammonium salt cationic surfactant or a polyethylene glycol type nonionic surfactant.

Specifically, the surfactant can be at a concentration of 0.1-7.0g/L (e.g., 0.1g/L, 1.0g/L, 3.0g/L, or 7.0g/L), the buffer can be at a concentration of 2.0-10.0g/L (e.g., 2.0g/L, 4.0g/L, 8.0g/L, or 10.0g/L), the cosolvent can be at a concentration of 3.0-15.0g/L (e.g., 3.0g/L, 5.0g/L, 10.0g/L, or 15.0g/L), and the preservative can be at a concentration of 0.1-3.0g/L (e.g., 0.1g/L, 0.5g/L, 1.0g/L, or 3.0 g/L).

The surfactant can be a combination of an amino acid type surfactant and a polyethylene glycol type nonionic surfactant, wherein the concentration of the amino acid type surfactant can be 0.1-1.0g/L (such as 0.1g/L, 0.5g/L or 1.0g/L), and the concentration of the polyethylene glycol type nonionic surfactant can be 0.3-5.0g/L (such as 0.3g/L, 1.5g/L or 5.0 g/L).

The surfactant may also be a combination of an amino acid type surfactant and a quaternary ammonium salt cationic surfactant, wherein the concentration of the amino acid type surfactant may be 0.1-1.0g/L (e.g., 0.1g/L, 0.5g/L, or 1.0g/L) and the concentration of the quaternary ammonium salt cationic surfactant may be 0.1-5.0g/L (e.g., 0.1g/L, 0.5g/L, or 5.0 g/L).

The surfactant may also be a combination of an amino acid type surfactant, a quaternary ammonium salt cationic surfactant, and a polyethylene glycol type nonionic surfactant, wherein the concentration of the amino acid type surfactant may be 0.1 to 1.0g/L (e.g., 0.1g/L, 0.5g/L, or 1.0g/L), the concentration of the quaternary ammonium salt cationic surfactant may be 0.01 to 5.0g/L (e.g., 0.3g/L, 0.5g/L, or 1.0g/L), and the concentration of the polyethylene glycol type nonionic surfactant may be 0.3 to 1.0g/L (e.g., 0.3g/L, 0.5g/L, or 1.0 g/L).

S102: the surfactant, buffer, cosolvent and preservative are poured into a dosing barrel filled with water.

S103: stirring to completely dissolve the surfactant, buffer, cosolvent, and preservative.

S104: filtering to obtain five-classification hemolytic agent for leukocyte.

Wherein, the concentration of the preservative in the leukocyte five-classification hemolytic agent is 0.1-3.0 g/L. The concentration of the surface active agent is 0.1-5.0g/L, the concentration of the buffering agent is 2.0-10.0g/L, and the concentration of the cosolvent is 3.0-15.0 g/L.

Specifically, 0.1-7.0g of surfactant, 2.0-10.0g of buffer, 3.0-15.0g of cosolvent and 0.1-3.0g of preservative can be weighed, and the surfactant, the buffer, the cosolvent and the preservative are poured into a mixing barrel filled with water to prepare 1L of leukocyte five-classification hemolytic agent solution.

The application provides a blood analyzer, this blood analyzer includes: a hemolytic agent bottle and a hemolytic agent injector communicated with the hemolytic agent bottle. The hemolytic agent bottle is used for containing the leukocyte five-classification hemolytic agent, and the hemolytic agent injector is used for sucking the hemolytic agent from the hemolytic agent bottle and pushing the leukocyte five-classification hemolytic agent into the reaction tank so as to mix the leukocyte five-classification hemolytic agent with the blood sample to be analyzed.

Specifically, the blood analyzer may include: the sample injector is respectively communicated with the sampling needle and the reaction tank. The sample injector is used for taking a blood sample to be detected through the sampling needle and adding the blood sample to be detected into the reaction pool.

The blood analyzer further includes: a hemolytic agent bottle and a hemolytic agent injector communicated with the hemolytic agent bottle. The hemolytic agent injector is communicated with the hemolytic agent bottle and used for sucking the leukocyte five-classification hemolytic agent from the hemolytic agent bottle and pushing the leukocyte five-classification hemolytic agent into the reaction pool to react with the blood sample to be detected. Furthermore, the sample injector is also used for pushing a sample flow formed after the blood sample to be detected enters the reaction pool and is reacted into the flow chamber for detection.

For the details of the leukocyte five-classification hemolytic agent in this embodiment, please refer to the leukocyte five-classification hemolytic agent in the above embodiment, which is not described herein again.

The application provides a detection method based on the blood analyzer, which comprises the following steps:

s201: and sucking the blood sample to be detected into the reaction pool.

Specifically, a blood sample to be detected is taken through a sampling needle and added into the reaction cell.

S202: and sucking the leukocyte five-classification hemolytic agent from the hemolytic agent bottle through a hemolytic agent syringe, and pushing the leukocyte five-classification hemolytic agent into the reaction tank.

Specifically, the hemolytic agent injector is communicated with the hemolytic agent bottle, and the hemolytic agent injector sucks the leukocyte five-classification hemolytic agent from the hemolytic agent bottle and pushes the leukocyte five-classification hemolytic agent into the reaction tank, so that the leukocyte five-classification hemolytic agent reacts with the blood sample to be detected.

S203: mixing the leukocyte penta-classification hemolytic agent with the blood sample to be detected.

S204: and detecting the size and morphological information of the white blood cells through a white blood cell channel measuring module.

Specifically, the leukocyte channel measurement module divides into: the four-classification measurement and BASO (basophilic granulocyte) measurement of the white blood cells, the five-classification hemolytic agent of the white blood cells is distributed into the reaction tank through the distribution system, and the blood sample to be detected is pushed into the reaction tank through the sampling needle to react. The white blood cell four-classification channel measuring module and the white blood cell BASO measuring module are carried out in parallel.

The present application is further described below with reference to examples:

example 1

The preparation method of the leukocyte five-classification hemolytic agent 1 comprises the following steps:

step (1): by weight, 0.2g of sodium lauroyl glutamate, 0.3g of cocamidopropyl dimethylamine lactate, 8.0g of acetic acid/potassium acetate buffer and 3.0g of ethylene glycol are weighed.

Step (2): sodium lauroyl glutamate, cocamidopropyl dimethylamine lactate, acetic acid/potassium acetate buffer and ethylene glycol were completely dissolved in water, and water was added to a constant volume of 1L.

And (3): filtering to obtain the five-classification hemolytic agent 1 for leucocyte.

In the leukocyte five-class hemolytic agent 1, the concentration of sodium lauroyl glutamate was 0.2g/L, the concentration of cocamidopropyl dimethylamine lactate was 0.3g/L, the concentration of acetic acid/potassium acetate was 8.0g/L, and the concentration of ethylene glycol was 3.0 g/L.

Example 2

The preparation method of the leukocyte penta-classification hemolytic agent 2 comprises the following steps:

step (1): weighing 0.1g of sodium cocoyl sarcosinate, 5.0g of hydroxypropyl lauryl dimethyl ammonium chloride, 10.0g of tris/hydrochloric acid buffer solution and 3.0g of methanol.

Step (2): completely dissolving sodium cocoyl sarcosinate, hydroxypropyl lauryl dimethyl ammonium chloride, tris/hydrochloric acid buffer solution and methanol in water, and adding water to a constant volume of 1L.

And (3): filtering to obtain leukocyte five-classification hemolytic agent 2.

In the leukocyte five-class hemolytic agent 2, the concentration of sodium cocoyl sarcosinate was 0.1g/L, the concentration of hydroxypropyl lauryl dimethyl ammonium chloride was 5.0g/L, the concentration of tris/hcl buffer solution was 10.0g/L, and the concentration of methanol was 10 g/L.

Example 3

The preparation method of the leukocyte five-classification hemolytic agent 3 comprises the following steps:

step (1): by weight, 0.1g sodium cocoyl sarcosinate, 5.0g lauryl polyoxyethylene (7) ether, 10.0g tris/hcl buffer, 3.0g methanol, and 3.0g glycerol were weighed.

Step (2): sodium cocoyl sarcosinate, lauryl polyoxyethylene (7) ether, tris/hcl buffer and methanol were completely dissolved in water, and water was added to a constant volume of 1L.

And (3): filtering to obtain the leukocyte five-classification hemolytic agent 3.

In the hemolyzing agent 3, the concentration of sodium cocoyl sarcosinate was 0.1g/L, the concentration of laureth-7 was 5.0g/L, the concentration of tris/hcl buffer was 10.0g/L, the concentration of methanol was 3.0g/L, and the concentration of glycerol was 3.0 g/L.

Example 4

The preparation method of the leukocyte penta-classification hemolytic agent 4 comprises the following steps:

step (1): 0.5g of sodium lauroyl sarcosinate, 1.0g of lauryl polyoxyethylene (7) ether, 2.0g of 3- (N-morpholinyl) propanesulfonic acid sodium salt/hydrochloric acid buffer solution, 8.0g of methanol and 3.0g of glycerol are weighed according to weight.

Step (2): completely dissolving sodium lauroyl sarcosinate, lauryl polyoxyethylene (7) ether, 3- (N-morpholinyl) propanesulfonic acid sodium salt/hydrochloric acid buffer solution, methanol and glycerol in water, and adding water to constant volume to 1L.

And (3): filtering to obtain leukocyte five-classification hemolytic agent 4.

In the leukocyte five-class hemolytic agent 4, the concentration of sodium lauroyl sarcosinate was 0.5g/L, the concentration of laureth-7 was 1.0g/L, the concentration of 3- (N-morpholino) propanesulfonic acid sodium salt/hydrochloric acid buffer solution was 2.0g/L, the concentration of methanol was 8.0g/L, and the concentration of glycerol was 3.0 g/L.

Example 5

The preparation method of the leukocyte five-classification hemolytic agent 5 comprises the following steps:

step (1): weighing 0.5g of sodium lauroyl sarcosinate, 4.0g of hydroxyethyl lauryl dimethyl ammonium chloride, 1.0g of lauryl polyoxyethylene (7) ether, 2g of 3- (N-morpholinyl) propanesulfonic acid sodium salt/hydrochloric acid buffer solution, 8g of methanol and 3.0g of glycerol according to weight.

Step (2): completely dissolving sodium lauroyl sarcosinate, hydroxyethyl lauryl dimethyl ammonium chloride, lauryl polyoxyethylene (7) ether, 3- (N-morpholinyl) propanesulfonic acid sodium salt/hydrochloric acid buffer solution, methanol and glycerol in water, and adding water to a constant volume of 1L.

And (3): filtering to obtain the five-classification hemolytic agent 5 for the white blood cells.

Wherein, in the leukocyte five-class hemolytic agent 5, the concentration of sodium lauroyl sarcosinate is 0.5g/L, the concentration of hydroxyethyl lauryl dimethyl ammonium chloride is 4.0g/L, the concentration of lauryl polyoxyethylene (7) ether is 1.0g/L, the concentration of 3- (N-morpholinyl) propanesulfonic acid sodium salt/hydrochloric acid buffer solution is 2g/L, the concentration of methanol is 8g/L, and the concentration of glycerol is 3.0 g/L.

Example 6

The preparation method of the leukocyte five-classification hemolytic agent 6 comprises the following steps:

step (1): weighing 0.1g of sodium cocoyl sarcosinate, 4.5g of dodecyl dimethyl benzyl ammonium chloride, 2.0g of polyethylene glycol p-isooctyl phenyl ether, 7.0g of phosphoric acid buffer and 12g of isopropanol.

Step (2): fully dissolving sodium cocoyl sarcosinate, dodecyl dimethyl benzyl ammonium chloride, polyethylene glycol p-isooctyl phenyl ether, phosphoric acid buffer pair and isopropanol in water, and adding water to a constant volume of 1L.

And (3): filtering to obtain the five-classification hemolytic agent 6 for leucocyte.

Wherein, in the leukocyte five-classification hemolytic agent 6, the concentration of sodium cocoyl sarcosinate is 0.1g/L, the concentration of dodecyl dimethyl benzyl ammonium chloride is 4.5g/L, the concentration of polyethylene glycol p-isooctyl phenyl ether is 2.0g/L, the concentration of phosphoric acid buffer pair is 7.0g/L, and the concentration of isopropanol is 12 g/L.

Example 7

The preparation method of the leukocyte five-classification hemolytic agent 7 comprises the following steps:

step (1): by weight, 0.2g of sodium lauroyl glutamate, 0.3g of cocamidopropyl dimethylamine lactate, 8.0g of acetic acid/potassium acetate buffer, 3.0g of ethylene glycol and 0.3g of Kethon are weighed.

Step (2): the sodium lauroyl glutamate, the cocamidopropyl dimethylamine lactate, the acetic acid/potassium acetate buffer solution, the ethylene glycol and the Kethon are completely dissolved in water, and water is added to the mixture to be constant volume of 1L.

And (3): filtering to obtain the five-classification hemolytic agent 7 for leucocyte.

Wherein, in the leukocyte five-classification hemolytic agent 7, the concentration of sodium lauroyl glutamate is 0.2g/L, the concentration of cocamidopropyl dimethylamine lactate is 0.3g/L, the concentration of acetic acid/potassium acetate buffer solution is 8.0g/L, the concentration of ethylene glycol is 3.0g/L, and the concentration of Kethon is 0.3 g/L.

Example 8

The preparation method of the leukocyte five-classification hemolytic agent 8 comprises the following steps:

step (1): weighing 0.1g of sodium cocoyl sarcosinate, 5.0g of hydroxypropyl lauryl dimethyl ammonium chloride, 10.0g of tris/hydrochloric acid buffer solution, 10g of methanol, 2.0g of potassium sorbate and 0.1g of benzoic acid.

Step (2): completely dissolving sodium cocoyl sarcosinate, hydroxypropyl lauryl dimethyl ammonium chloride, tris/hydrochloric acid buffer solution, methanol, potassium sorbate and benzoic acid in water, and adding water to a constant volume of 1L.

And (3): filtering to obtain the five-classification hemolytic agent 8 for leucocyte.

In the leukocyte five-class hemolytic agent 8, the concentration of sodium cocoyl sarcosinate was 0.1g/L, the concentration of hydroxypropyl lauryl dimethyl ammonium chloride was 5.0g/L, the concentration of tris/hydrochloric acid buffer was 10.0g, the concentration of methanol was 10g/L, the concentration of potassium sorbate was 2.0g/L, and the concentration of benzoic acid was 0.1 g/L.

Example 9

The preparation method of the leukocyte five-classification hemolytic agent 9 comprises the following steps:

step (1): weighing 0.5g of sodium lauroyl sarcosinate, 4.0g of hydroxyethyl lauryl dimethyl ammonium chloride, 1.0g of lauryl polyoxyethylene (7) ether, 2.0g of 3- (N-morpholinyl) propanesulfonic acid sodium salt/hydrochloric acid buffer solution, 8g of methanol, 3.0g of glycerol, 1.0g of Kethon and 0.1g of benzoic acid.

Step (2): completely dissolving sodium lauroyl sarcosinate, hydroxyethyl lauryl dimethyl ammonium chloride, lauryl polyoxyethylene (7) ether, 3- (N-morpholinyl) propanesulfonic acid sodium salt/hydrochloric acid buffer solution, methanol, glycerol, kaempferol and benzoic acid in water, and adding water to a constant volume of 1L.

And (3): filtering to obtain the five-classification hemolytic agent 9 for leucocyte.

Wherein, in the leukocyte five-classification hemolytic agent 9, the concentration of sodium lauroyl sarcosinate is 0.5g/L, the concentration of hydroxyethyl lauryl dimethyl ammonium chloride is 4.0g/L, the concentration of lauryl polyoxyethylene (7) ether is 1.0g/L, the concentration of 3- (N-morpholinyl) propanesulfonic acid sodium salt/hydrochloric acid buffer solution is 2g/L, the concentration of methanol is 8g/L, the concentration of glycerol is 3.0g/L, the concentration of Kjeldahl is 1.0g/L, and the concentration of benzoic acid is 0.1 g/L.

Example 10

The preparation method of the five-classification hemolytic agent 10 for the white blood cells comprises the following steps:

step (1): weighing 0.1g of sodium cocoyl sarcosinate, 4.5g of dodecyl dimethyl benzyl ammonium chloride, 2.0g of polyethylene glycol p-isooctyl phenyl ether, 7.0g of phosphoric acid buffer, 12.0g of isopropanol, 0.2g of Kethon and 3.0g of potassium sorbate.

Step (2): fully dissolving sodium cocoyl sarcosinate, dodecyl dimethyl benzyl ammonium chloride, polyethylene glycol p-isooctyl phenyl ether, phosphoric acid buffer pair, isopropanol, Kethon and potassium sorbate in water, and adding water to a constant volume of 1L.

And (3): filtering to obtain the five-classification hemolytic agent 10 for leucocyte.

Wherein, in the leukocyte five-classification hemolytic agent 10, the concentration of sodium cocoyl sarcosinate is 0.1g/L, the concentration of dodecyl dimethyl benzyl ammonium chloride is 4.5g/L, the concentration of polyethylene glycol p-isooctyl phenyl ether is 2.0g/L, the concentration of phosphoric acid buffer pair is 7.0g/L, the concentration of isopropanol is 12.0g/L, Kjen is 0.2g/L, and the concentration of potassium sorbate is 3.0 g/L.

Example 11

On a hematology analyzer, the results of the detection of the blood sample to be tested of volunteer a in the hematology analyzer using the three batches of the five leukocyte classification hemolyzing agent 1 prepared in example 1 of the present application are shown in table 1, wherein WBC represents the leukocyte concentration, Neu% represents the neutrophil concentration, Lym% represents the lymphocyte concentration, Mon% represents the monocyte concentration, Eos represents the eosinophil concentration, and Bas% represents the basophil concentration. Please refer to steps S301 to S204 of the above embodiments, which are not described herein.

Wherein, the standard value of the detection item is: the method comprises the following steps of detecting a blood sample to be detected of a volunteer A in a blood analyzer by adopting an industrially universal leucocyte classification hemolytic agent, wherein the industrially universal leucocyte classification hemolytic agent comprises the following components:

reagent 1: an ethidine-acridine heterodimer, ethylene glycol;

reagent 2: phthalic acid, quaternary ammonium salt surfactant, and pyridine

Salt type surfactants, nonionic surfactants.

TABLE 1 leukocyte penta-Classification test results of three batches of leukocyte penta-Classification hemolytic agent 1

Example 12

The results of 10 tests performed on a blood sample to be tested of volunteer a in a hematology analyzer using the leukocyte five-classification hemolyzing agent 1 prepared in example 1 of the present application are shown in table 2, wherein WBC represents a leukocyte concentration, Neu% represents a neutrophil concentration, Lym% represents a lymphocyte concentration, Mon% represents a monocyte concentration, Eos% represents an eosinophil concentration, and Bas% represents a basophil concentration. Please refer to steps S301 to S204 of the above embodiments, which are not described herein.

TABLE 2 leukocyte five-Classification hemolytic agent 1 results of 10 leukocyte five-Classification detection

Example 13

On a hematology analyzer, the results of the blood sample to be tested of volunteer B in the hematology analyzer using the three batches of the leukocyte five-classification hemolytic agent 2 prepared in example 2 of the present application are shown in table 3, wherein WBC represents the leukocyte concentration, Neu% represents the neutrophil concentration, Lym% represents the lymphocyte concentration, Mon% represents the monocyte concentration, Eos% represents the eosinophil concentration, and Bas% represents the basophil concentration. Please refer to steps S301 to S204 of the above embodiments, which are not described herein.

Wherein, the standard value of the detection item is: the method comprises the following steps of detecting a blood sample to be detected of a volunteer B in a blood analyzer by adopting an industrially universal leucocyte classification hemolytic agent, wherein the industrially universal leucocyte classification hemolytic agent comprises the following components:

reagent 1: an ethidine-acridine heterodimer, ethylene glycol;

reagent 2: phthalic acid, quaternary ammonium salt surfactant, and pyridine

Salt type surfactants, nonionic surfactants.

TABLE 3 white blood cell five-classification test results of three batches of white blood cell five-classification hemolytic agent 2

Example 14

The results of 10 measurements on a blood sample to be tested of volunteer B on a hematology analyzer using the leukocyte five-classification hemolytic agent 2 prepared in example 2 of the present application are shown in table 4, wherein WBC represents a leukocyte concentration, Neu% represents a neutrophil concentration, Lym% represents a lymphocyte concentration, Mon% represents a monocyte concentration, Eos% represents an eosinophil concentration, and Bas% represents a basophil concentration. Please refer to steps S301 to S204 of the above embodiments, which are not described herein.

TABLE 4 leukocyte penta-Classification hemolytic agent 2 10 times leukocyte penta-Classification test results

Example 15

On a hematology analyzer, the results of the blood sample to be tested of volunteer B in the hematology analyzer using the three batches of the five leukocyte classification hemolyzing agent 3 prepared in example 3 of the present application are shown in table 5, wherein WBC represents the leukocyte concentration, Neu% represents the neutrophil concentration, Lym% represents the lymphocyte concentration, Mon% represents the monocyte concentration, Eos% represents the eosinophil concentration, and Bas% represents the basophil concentration. Please refer to steps S301 to S204 of the above embodiments, which are not described herein.

reagent 1: an ethidine-acridine heterodimer, ethylene glycol;

reagent 2: phthalic acid, quaternary ammonium salt surfactant, and pyridine

Salt type surfactants, nonionic surfactants.

TABLE 5 leukocyte five-Classification test results of three batches of leukocyte five-Classification hemolytic agent 3

Example 16

The results of 10 measurements on a blood sample to be tested of volunteer B on a hematology analyzer using the leukocyte five-classification hemolytic agent 3 prepared in example 3 of the present application are shown in table 6, wherein WBC represents a leukocyte concentration, Neu% represents a neutrophil concentration, Lym% represents a lymphocyte concentration, Mon% represents a monocyte concentration, Eos% represents an eosinophil concentration, and Bas% represents a basophil concentration. Please refer to steps S301 to S204 of the above embodiments, which are not described herein.

TABLE 6 leukocyte five-Classification hemolytic agent 3 results of 10 leukocyte five-Classification detection

Example 17

On a hematology analyzer, the results of the blood sample to be tested of volunteer B in the hematology analyzer using the three batches of leukocyte penta-sorting hemolytic agent 4 prepared in example 4 of the present application are shown in table 7, wherein WBC indicates leukocyte concentration, Neu% indicates neutrophil concentration, Lym% indicates lymphocyte concentration, Mon% indicates monocyte concentration, Eos% indicates eosinophil concentration, and Bas% indicates basophil concentration. Please refer to steps S301 to S204 of the above embodiments, which are not described herein.

reagent 1: an ethidine-acridine heterodimer, ethylene glycol;

reagent 2: phthalic acid, quaternary ammonium salt surfactant, and pyridine

Salt type surfactants, nonionic surfactants.

TABLE 7 leukocyte five-Classification test results of three batches of leukocyte five-Classification hemolytic agent 4

Example 18

The results of 10 measurements of the blood sample to be tested of volunteer B on a hematology analyzer using the leukocyte five-classification hemolyzing agent 4 prepared in example 4 of the present application are shown in table 8, wherein WBC represents the leukocyte concentration, Neu% represents the neutrophil concentration, Lym% represents the lymphocyte concentration, Mon% represents the monocyte concentration, Eos% represents the eosinophil concentration, and Bas% represents the basophil concentration. Please refer to steps S301 to S204 of the above embodiments, which are not described herein.

TABLE 8 leukocyte penta-classification hemolytic agent 4 10 times leukocyte penta-classification detection results

Example 19

On a hematology analyzer, the results of the blood samples to be tested of volunteer C in the hematology analyzer using the three batches of the five leukocyte classification hemolyzing agent 5 prepared in example 5 of the present application are shown in table 9, wherein WBC represents the leukocyte concentration, Neu% represents the neutrophil concentration, Lym% represents the lymphocyte concentration, Mon% represents the monocyte concentration, Eos% represents the eosinophil concentration, and Bas% represents the basophil concentration. Please refer to steps S301 to S204 of the above embodiments, which are not described herein.

Wherein, the standard value of the detection item is: detecting a detection result obtained after detecting a blood sample to be detected of a volunteer C in a blood analyzer by adopting a leukocyte classification hemolytic agent universal to the industry, wherein the leukocyte classification hemolytic agent universal to the industry comprises the following components:

reagent 1: an ethidine-acridine heterodimer, ethylene glycol;

reagent 2: phthalic acid, quaternary ammonium salt surfactant, and pyridine

Salt type surfactants, nonionic surfactants.

TABLE 9 leukocyte five-Classification test results of three batches of leukocyte five-Classification hemolytic agent 5

Example 20

The results of 10 tests on a blood sample to be tested of volunteer C in a hematology analyzer using the leukocyte five-classification hemolytic agent 5 prepared in example 5 of the present application are shown in table 10, wherein WBC represents a leukocyte concentration, Neu% represents a neutrophil concentration, Lym% represents a lymphocyte concentration, Mon% represents a monocyte concentration, Eos% represents an eosinophil concentration, and Bas% represents a basophil concentration. Please refer to steps S301 to S204 of the above embodiments, which are not described herein.

TABLE 10 leukocyte five-Classification hemolytic agent 5 results of 10 leukocyte five-Classification detection

Example 21

On a hematology analyzer, the results of the blood samples to be tested of volunteer D in the hematology analyzer using the three batches of the five leukocyte classification hemolyzing agents 6 prepared in example 6 of the present application are shown in table 11, wherein WBC represents the leukocyte concentration, Neu% represents the neutrophil concentration, Lym% represents the lymphocyte concentration, Mon% represents the monocyte concentration, Eos% represents the eosinophil concentration, and Bas% represents the basophil concentration. Please refer to steps S301 to S204 of the above embodiments, which are not described herein.

Wherein, the standard value of the detection item is: the method comprises the following steps of detecting a blood sample to be detected of a volunteer D in a blood analyzer by adopting an industrially universal leucocyte classification hemolytic agent, wherein the industrially universal leucocyte classification hemolytic agent comprises the following components:

reagent 1: an ethidine-acridine heterodimer, ethylene glycol;

reagent 2: phthalic acid, quaternary ammonium salt surfactant, and pyridine

Salt type surfactants, nonionic surfactants.

TABLE 11 leukocyte penta-Classification test results of three batches of leukocyte penta-Classification hemolytic agent 6

Example 22

The results of 10 tests on a blood sample to be tested of volunteer D in a hematology analyzer using the leukocyte five-classification hemolyzing agent 6 prepared in example 6 of the present application are shown in table 12, wherein WBC represents a leukocyte concentration, Neu% represents a neutrophil concentration, Lym% represents a lymphocyte concentration, Mon% represents a monocyte concentration, Eos% represents an eosinophil concentration, and Bas% represents a basophil concentration. Please refer to steps S301 to S204 of the above embodiments, which are not described herein.

TABLE 12 leukocyte five-Classification hemolytic agent 6 results of 10 leukocyte five-Classification test

Example 23

The results of the measurement of the blood sample to be tested of volunteer a on the hematology analyzer using three batches of the five-classification hemolyzing agent 7 prepared according to example 7 of the present application, which was shipped from 12 months, are shown in table 7, where WBC represents the leukocyte concentration, Neu% represents the neutrophil concentration, Lym% represents the lymphocyte concentration, Mon% represents the monocyte concentration, Eos represents the eosinophil concentration, and Bas% represents the basophil concentration. Please refer to steps S301 to S204 of the above embodiments, which are not described herein.

reagent 1: an ethidine-acridine heterodimer, ethylene glycol;

reagent 2: phthalic acid, quaternary ammonium salt surfactant, and pyridine

Salt type surfactants, nonionic surfactants.

TABLE 13 leukocyte five-Classification test results of three batches of leukocyte five-Classification hemolytic agent 7

Example 24

The results of 10 tests on a blood sample to be tested of volunteer a in a blood analyzer using the leukocyte five-classification hemolytic agent 7 prepared in example 7 of the present application shipped from 12 months on the blood analyzer are shown in table 14, in which WBC represents a leukocyte concentration, Neu% represents a neutrophil concentration, Lym% represents a lymphocyte concentration, Mon% represents a monocyte concentration, Eos represents an eosinophil concentration, and Bas% represents a basophil concentration.

TABLE 14 leukocyte five-Classification hemolytic agent 7 10 times leukocyte five-Classification test results

Example 25

The results of the measurement of the blood sample to be tested of the volunteer B on the hematology analyzer using the three batches of the five-classification hemolyzing agent 8 prepared in example 8 of the present application, which was shipped from 12 months, are shown in table 15, where WBC represents the leukocyte concentration, Neu% represents the neutrophil concentration, Lym% represents the lymphocyte concentration, Mon% represents the monocyte concentration, Eos represents the eosinophil concentration, and Bas% represents the basophil concentration. Please refer to steps S301 to S204 of the above embodiments, which are not described herein.

reagent 1: an ethidine-acridine heterodimer, ethylene glycol;

reagent 2: phthalic acid, quaternary ammonium salt surfactant, and pyridine

Salt type surfactants, nonionic surfactants.

TABLE 15 leukocyte five-Classification test results of three batches of leukocyte five-Classification hemolytic agent 8

Example 26

The results of 10 tests performed on a blood sample to be tested of volunteer B in a blood analyzer using the five-classification hemolyzing agent for leukocytes 8 prepared in example 8 of the present application, which was shipped from 12 months in the factory, are shown in table 16, in which WBC represents the leukocyte concentration, Neu% represents the neutrophil concentration, Lym% represents the lymphocyte concentration, Mon% represents the monocyte concentration, Eos% represents the eosinophil concentration, and Bas% represents the basophil concentration.

TABLE 16 leukocyte five-Classification hemolytic agent 8 results of 10 leukocyte five-Classification detection

Example 27

The results of the measurement of the blood sample to be tested of the volunteer C on the hematology analyzer using the three batches of the five-classification hemolyzing agent 9 prepared in example 9 of the present application, which was shipped from the factory for 12 months, are shown in table 17, where WBC represents the leukocyte concentration, Neu% represents the neutrophil concentration, Lym% represents the lymphocyte concentration, Mon% represents the monocyte concentration, Eos% represents the eosinophil concentration, and Bas% represents the basophil concentration. Please refer to steps S301 to S204 of the above embodiments, which are not described herein.

reagent 1: an ethidine-acridine heterodimer, ethylene glycol;

reagent 2: phthalic acid, quaternary ammonium salt surfactant, and pyridine

Salt type surfactants, nonionic surfactants.

TABLE 17 leukocyte five-Classification test results of three batches of leukocyte five-Classification hemolytic agent 9

Example 28

The results of 10 tests on a blood sample to be tested of volunteer C in a blood analyzer using the five-classification hemolyzing agent for leukocytes 9 prepared in example 9 of the present application, which was shipped from 12 months, are shown in table 18, where WBC represents a leukocyte concentration, Neu% represents a neutrophil concentration, Lym% represents a lymphocyte concentration, Mon% represents a monocyte concentration, Eos represents an eosinophil concentration, and Bas% represents a basophil concentration. Please refer to steps S301 to S204 of the above embodiments, which are not described herein.

TABLE 18 leukocyte five-Classification hemolytic agent 9 results of 10 leukocyte five-Classification detection

Example 29

The results of the measurement of the blood sample to be tested of volunteer D on the hematology analyzer using three batches of the five-classification hemolyzing agent 10 of leukocyte prepared according to example 10 of the present application, which was prepared at 12 months of factory, are shown in table 19, where WBC represents the leukocyte concentration, Neu% represents the neutrophil concentration, Lym% represents the lymphocyte concentration, Mon% represents the monocyte concentration, Eos% represents the eosinophil concentration, and Bas% represents the basophil concentration. Please refer to steps S301 to S204 of the above embodiments, which are not described herein.

reagent 1: an ethidine-acridine heterodimer, ethylene glycol;

reagent 2: phthalic acid, quaternary ammonium salt surfactant, and pyridine

Salt type surfactants, nonionic surfactants.

TABLE 19 leukocyte five-Classification test results of three batches of leukocyte five-Classification hemolytic agent 10

Example 30

The results of 10 tests performed on a blood sample to be tested of volunteer D in a blood analyzer using the five-classification hemolyzing agent for leukocytes 10 prepared in example 10 of the present application, which was shipped from 12 months in the factory, are shown in table 20, in which WBC represents the leukocyte concentration, Neu% represents the neutrophil concentration, Lym% represents the lymphocyte concentration, Mon% represents the monocyte concentration, Eos% represents the eosinophil concentration, and Bas% represents the basophil concentration. Please refer to steps S301 to S204 of the above embodiments, which are not described herein.

TABLE 20 five-Classification of leukocytes hemolytic agent 10 results of 10 five-Classification of leukocytes

As can be seen from tables 1-20 above:

(1) in examples 11 to 30, when the hematology analyzer is running, only a single addition of the leukocyte five-classification hemolytic agent of the present invention is needed, and no additional hemolytic agent B or hemolytic agent C is needed, so that the red blood cells can be lysed and the leukocytes can be stabilized, and compared with the detection result of the standard value, the absolute deviation and CV% of the detection result of the above examples are within the range of the standard absolute deviation and the standard CV%, so that the total number of leukocytes, the leukocyte five-classification count and other parameters can be determined after the leukocyte five-classification hemolytic agent 1 to 10 of the present invention is mixed with the blood sample to be detected, thereby reducing the use cost and reducing the detection steps.

(2) In examples 1-2 and 11-14, amino acid type surfactants and quaternary ammonium salt cationic surfactants were selected as the surfactants, and the damage of the quaternary ammonium salt cationic surfactants to the structure of leukocytes can be effectively avoided by strictly controlling the amount of the quaternary ammonium salt cationic surfactants (the concentration of the quaternary ammonium salt cationic surfactants is 7.0g/L or less).

(3) In examples 3-4 and 15-18, the combination of amino acid type surfactant and polyethylene glycol type nonionic surfactant was used as the surfactant, and the structure of leukocytes can be effectively prevented from being damaged by the quaternary ammonium salt cationic surfactant without using the quaternary ammonium salt cationic surfactant, and the detection purpose can be achieved.

(4) In examples 5 to 6 and 19 to 22, the amino acid type surfactant, the polyethylene glycol type nonionic surfactant, and the quaternary ammonium salt cationic surfactant were used as the surfactants, and in the case of using the three surfactants in combination, the detection object was achieved, and at the same time, the amount of the quaternary ammonium salt cationic surfactant (the concentration of the quaternary ammonium salt cationic surfactant was 5.0g/L or less) was strictly controlled, and the damage of the structure of leukocytes by the quaternary ammonium salt cationic surfactant was effectively prevented.

(5) In examples 7 to 9, after the preservative is added to the leukocyte five-classification hemolytic agent, the reagent can be prevented from deteriorating, the storage and use time of the reagent can be prolonged, the performance of the reagent can be still stable within 12 months after the reagent leaves a factory, and the effective period of the leukocyte five-classification hemolytic agent product can be prolonged.

The above description is only for the purpose of illustrating embodiments of the present application and is not intended to limit the scope of the present application, and all modifications of equivalent structures and equivalent processes, which are made by the contents of the specification and the drawings of the present application or are directly or indirectly applied to other related technical fields, are also included in the scope of the present application.

Claims

1. A leukocyte penta-sorting hemolytic agent, comprising: surfactants, buffers and cosolvents;

2. The hemolyzing agent for five white blood cell classification according to claim 1, further comprising: and (4) a preservative.

3. The five-classification hemolytic agent for leukocyte according to claim 2, wherein the concentration of the surfactant is 0.1-7.0g/L, the concentration of the buffer is 2.0-10.0g/L, the concentration of the cosolvent is 3.0-15.0g/L, and the concentration of the preservative is 0.1-3.0 g/L.

4. The hemolyzing agent for leukocyte pentaclassification according to claim 1,

the amino acid type surfactant comprises at least one of sodium lauroyl glutamate, sodium lauroyl sarcosinate, sodium cocoyl glycinate, cocoyl triethanolamine glutamate, sodium palmitoyl glutamate, cocoyl propyl glutamate, disodium cocoyl glutamate, sodium cocoyl glutamate, N-myristyl-L-glutamic acid dibutylamide, or sodium octadecyl dimethyleneaminodicarboxylate;

the quaternary ammonium salt cationic surfactant comprises at least one of cocamidopropyl dimethyl amine lactate, hydroxyethyl lauryl dimethyl ammonium chloride, dodecyl dimethyl benzyl ammonium chloride, tetradecyl trimethyl ammonium chloride, hexadecyl trimethyl ammonium chloride, dodecyl trimethyl ammonium chloride, hexadecyl trimethyl ammonium bromide, dodecyl trimethyl ammonium bromide or tetradecyl trimethyl ammonium bromide;

the polyethylene glycol nonionic surfactant comprises at least one of lauryl polyoxyethylene (7) ether, polyethylene glycol p-isooctyl phenyl ether, polyoxyethylene fatty amide or stearic acid amide polyoxyethylene (6) ether.

5. The hemolyzing agent for leukocyte pentaclassification according to claim 1,

the buffer comprises at least one of acetic acid/potassium acetate buffer solution, tris/hydrochloric acid buffer solution, 3- (N-morpholinyl) propanesulfonic acid sodium salt/hydrochloric acid buffer solution, or disodium hydrogen phosphate/sodium dihydrogen phosphate buffer salt;

the cosolvent comprises at least one of ethylene glycol, methanol, ethanol, isopropanol or glycerol.

6. The five-classification hemolytic agent for leukocyte according to claim 2,

the preservative is at least one of kaempferol, potassium sorbate or benzoic acid.

7. A preparation method of a five-classification hemolytic agent for leucocytes is characterized by comprising the following steps:

weighing a surfactant, a buffering agent, a cosolvent and a preservative according to a preset formula, wherein the surfactant comprises an amino acid type surfactant and at least one of a quaternary ammonium salt cationic surfactant or a polyethylene glycol type nonionic surfactant;

pouring the surfactant, the buffer, the cosolvent and the preservative into a dosing barrel filled with water;

stirring to completely dissolve the surfactant, the buffer, the cosolvent, and the preservative;

filtering to obtain the leukocyte five-classification hemolytic agent;

wherein, in the leukocyte five-classification hemolytic agent, the concentration of the preservative is 0.1-3.0 g/L. The concentration of the surfactant is 0.1-7.0g/L, the concentration of the buffering agent is 2.0-10.0g/L, and the concentration of the cosolvent is 3.0-15.0 g/L.

8. The use of a five white blood cell sorting hemolytic agent according to any one of claims 1-6 for five white blood cell sorting counting.

9. A blood analyzer, comprising: a hemolytic agent bottle and a hemolytic agent injector communicated with the hemolytic agent bottle;

the hemolytic agent bottle is used for containing the leukocyte penta-classification hemolytic agent according to any one of claims 1-6, and the hemolytic agent injector is used for sucking the hemolytic agent from the hemolytic agent bottle and pushing the leukocyte penta-classification hemolytic agent into the reaction tank so as to mix the leukocyte penta-classification hemolytic agent with the blood sample to be analyzed.

10. A detection method based on the blood analyzer of claim 9, wherein the detection method comprises:

sucking a blood sample to be detected into a reaction pool;

sucking the leukocyte penta-sorting hemolytic agent of any one of claims 1-6 from the hemolytic agent bottle by a hemolytic agent syringe and pushing the leukocyte penta-sorting hemolytic agent into a reaction tank;

mixing the five-classification hemolytic agent for the white blood cells with the blood sample to be detected;

and detecting the size and morphological information of the white blood cells through a white blood cell channel measuring module.