CN108169468B - Diluent suitable for various blood analyzers and preparation method thereof - Google Patents
Diluent suitable for various blood analyzers and preparation method thereof Download PDFInfo
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- CN108169468B CN108169468B CN201711312375.1A CN201711312375A CN108169468B CN 108169468 B CN108169468 B CN 108169468B CN 201711312375 A CN201711312375 A CN 201711312375A CN 108169468 B CN108169468 B CN 108169468B
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- 239000003085 diluting agent Substances 0.000 title claims abstract description 63
- 210000004369 blood Anatomy 0.000 title claims abstract description 44
- 239000008280 blood Substances 0.000 title claims abstract description 44
- 238000002360 preparation method Methods 0.000 title abstract description 6
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims abstract description 27
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 claims abstract description 22
- 230000003204 osmotic effect Effects 0.000 claims abstract description 16
- 239000003381 stabilizer Substances 0.000 claims abstract description 14
- 229960004063 propylene glycol Drugs 0.000 claims abstract description 12
- 239000003146 anticoagulant agent Substances 0.000 claims abstract description 11
- 229940127219 anticoagulant drug Drugs 0.000 claims abstract description 11
- 239000000872 buffer Substances 0.000 claims abstract description 10
- 239000003755 preservative agent Substances 0.000 claims abstract description 10
- 230000002335 preservative effect Effects 0.000 claims abstract description 10
- 239000006172 buffering agent Substances 0.000 claims abstract description 4
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 22
- 239000011780 sodium chloride Substances 0.000 claims description 11
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims description 6
- IWZKICVEHNUQTL-UHFFFAOYSA-M potassium hydrogen phthalate Chemical compound [K+].OC(=O)C1=CC=CC=C1C([O-])=O IWZKICVEHNUQTL-UHFFFAOYSA-M 0.000 claims description 6
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 4
- CHHHXKFHOYLYRE-UHFFFAOYSA-M 2,4-Hexadienoic acid, potassium salt (1:1), (2E,4E)- Chemical compound [K+].CC=CC=CC([O-])=O CHHHXKFHOYLYRE-UHFFFAOYSA-M 0.000 claims description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical group OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims description 2
- 239000006173 Good's buffer Substances 0.000 claims description 2
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims description 2
- 229940071106 ethylenediaminetetraacetate Drugs 0.000 claims description 2
- 239000008363 phosphate buffer Substances 0.000 claims description 2
- 239000001103 potassium chloride Substances 0.000 claims description 2
- 235000011164 potassium chloride Nutrition 0.000 claims description 2
- 235000010241 potassium sorbate Nutrition 0.000 claims description 2
- 239000004302 potassium sorbate Substances 0.000 claims description 2
- 229940069338 potassium sorbate Drugs 0.000 claims description 2
- OTYBMLCTZGSZBG-UHFFFAOYSA-L potassium sulfate Chemical compound [K+].[K+].[O-]S([O-])(=O)=O OTYBMLCTZGSZBG-UHFFFAOYSA-L 0.000 claims description 2
- 229910052939 potassium sulfate Inorganic materials 0.000 claims description 2
- 235000011151 potassium sulphates Nutrition 0.000 claims description 2
- WXMKPNITSTVMEF-UHFFFAOYSA-M sodium benzoate Chemical compound [Na+].[O-]C(=O)C1=CC=CC=C1 WXMKPNITSTVMEF-UHFFFAOYSA-M 0.000 claims description 2
- 235000010234 sodium benzoate Nutrition 0.000 claims description 2
- 239000004299 sodium benzoate Substances 0.000 claims description 2
- 235000002639 sodium chloride Nutrition 0.000 claims description 2
- 229910052938 sodium sulfate Inorganic materials 0.000 claims description 2
- 235000011152 sodium sulphate Nutrition 0.000 claims description 2
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 29
- 238000013329 compounding Methods 0.000 abstract description 4
- 238000000338 in vitro Methods 0.000 abstract description 4
- 238000011161 development Methods 0.000 abstract description 3
- 238000005259 measurement Methods 0.000 abstract description 3
- 239000000126 substance Substances 0.000 abstract description 3
- 238000012827 research and development Methods 0.000 abstract description 2
- 210000000601 blood cell Anatomy 0.000 description 16
- 239000000243 solution Substances 0.000 description 13
- 238000001514 detection method Methods 0.000 description 11
- 238000004458 analytical method Methods 0.000 description 8
- 238000012360 testing method Methods 0.000 description 5
- 210000003743 erythrocyte Anatomy 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 3
- DNIAPMSPPWPWGF-GSVOUGTGSA-N (R)-(-)-Propylene glycol Chemical compound C[C@@H](O)CO DNIAPMSPPWPWGF-GSVOUGTGSA-N 0.000 description 2
- 102000001554 Hemoglobins Human genes 0.000 description 2
- 108010054147 Hemoglobins Proteins 0.000 description 2
- 239000013522 chelant Substances 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 238000005534 hematocrit Methods 0.000 description 2
- -1 iron ions Chemical class 0.000 description 2
- 210000000265 leukocyte Anatomy 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 235000013772 propylene glycol Nutrition 0.000 description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 230000004931 aggregating effect Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000023555 blood coagulation Effects 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000003864 humus Substances 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Substances [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/483—Physical analysis of biological material
- G01N33/487—Physical analysis of biological material of liquid biological material
- G01N33/49—Blood
- G01N33/491—Blood by separating the blood components
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Physics & Mathematics (AREA)
- Food Science & Technology (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- Ecology (AREA)
- Biophysics (AREA)
- Medicinal Chemistry (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention relates to the technical field of in-vitro diagnostic reagents, in particular to a diluent suitable for various blood analyzers and a preparation method thereof. The diluent comprises a diluent basic formula, 1.0-15.0 g/L of osmotic pressure compensation agent and 0.6-1.5 g/L of buffering agent; the basic formula of the diluent comprises 0.2-1.0 g/L of anticoagulant, 0.3-0.5 g/L of preservative and 0.2-1.5 g/L of stabilizer. The diluent for the blood analyzer improves the stability and the measurement reliability of a blood sample substance by compounding the stabilizer isopropanol and the 1, 2-propylene glycol in the formula, and can be suitable for the blood analyzers of different manufacturers by fixing the basic formula of the diluent and only adjusting the contents of the buffer and the osmotic pressure compensator, thereby simplifying the development steps of in-vitro diagnostic reagents and reducing the research and development cost. And breaks monopoly of reagents of original plants, and reduces the use cost of reagents of all levels of medical institutions.
Description
Technical Field
The invention relates to the technical field of in-vitro diagnostic reagents, in particular to a diluent suitable for various blood analyzers and a preparation method thereof.
Background
Blood cell analysis is the most common item in clinical examination, and blood cells in a blood sample are analyzed and detected by a blood analyzer by extracting the blood sample from a human body, and the detected parameters comprise White Blood Cells (WBC), Red Blood Cells (RBC), Hemoglobin (HGB), Platelets (PLT), Hematocrit (HCT), Mean Corpuscular Volume (MCV) and the like.
The blood analyzer uses a plurality of reagents to assist the detection, wherein a diluent is an indispensable reagent. The diluent for the blood analyzer is mainly used for diluting blood cells and preventing the blood cells from aggregating and adhering; and a certain PH value and osmotic pressure are maintained, so that a proper environment is provided for blood cell detection.
At present, reagents of blood analyzers in the market are various in varieties and are special for special machines, reagents of original factories are often expensive, particularly diluents, the daily work requirement is large, the application cost is very high, and the blood analyzers bring troubles on the cost to vast medical institutions. Therefore, the development of a diluent which has a simple formula and low cost, is suitable for various machine types, can replace the diluent used on the machine of the original factory reagent, and reduces the use cost of the diluent of the hematology analyzer with various levels of medical structures is necessary.
Disclosure of Invention
In order to solve the technical problems, the invention provides a diluent suitable for various blood analyzers and a preparation method thereof.
The technical scheme of the invention is as follows:
the invention provides a diluent suitable for various blood analyzers, which comprises a diluent basic formula, an osmotic pressure compensation agent 1.0-15.0 g/L and a buffering agent 0.6-1.5 g/L;
the basic formula of the diluent comprises 0.2-1.0 g/L of anticoagulant, 0.3-0.5 g/L of preservative and 0.2-1.5 g/L of stabilizer.
Further, the anticoagulant is ethylenediamine tetraacetate, and the concentration of the anticoagulant is 0.2-1.0 g/L.
Further, the anticoagulant is EDTA-K2。
Further, the preservative is one or more of sodium benzoate, potassium sorbate and potassium hydrogen phthalate, and the concentration of the preservative is 0.3-0.5 g/L; a preferred preservative is potassium hydrogen phthalate.
Further, the stabilizer is one or more of ethanol, ethylene glycol, isopropanol and 1, 2-propylene glycol; the preferable stabilizer is isopropanol and 1, 2-propylene glycol, wherein the concentration of the isopropanol is 0.1-2.0 g/L, and the preferable concentration is 0.5-1.5 g/L; the concentration of 1, 2-propanediol is 0.1 to 1.5g/L, preferably 0.2 to 1.2 g/L.
Further, the osmotic pressure compensator is one or more of sodium chloride, potassium chloride, sodium sulfate and potassium sulfate, and preferably sodium chloride; the concentration is 5.0-10.0 g/L.
Further, the buffer is one or more of phosphate buffer, Tris-HCl buffer and Good's buffer; preferably Tris-HCl buffer; the concentration is 0.6-1.5 g/L.
The pH value of the diluent prepared by the invention is 7.0-9.0.
In a preferred embodiment of the present invention, the diluent is prepared by determining the contents of anticoagulant, stabilizer and preservative, adjusting the buffer to maintain the pH of the reagent consistent with the factory reagent, and adjusting the osmotic pressure compensating agent to maintain the average red blood cell volume (MCV) of the on-machine test consistent with the factory reagent based on the on-machine results of the blood analyzer. The preparation process includes determining the basic recipe of diluent, and regulating osmotic pressure compensator and buffering agent to make the diluent match with corresponding blood analyzer.
The anticoagulant can chelate with some positive ions (especially calcium and iron ions) in the blood sample after being added to the blood sample to form stable chelate, thereby preventing blood coagulation. The preservative ensures that the substances are not humus or rotten, and further ensures the stability of the diluent. The stabilizing agent is used for maintaining the osmotic pressure and potential balance of the blood sample, so that the stability of each cell and component in the blood sample is improved, and meanwhile, the solution after the blood sample is diluted presents a stable hydroelectric layer, and the uniformity and stability of the whole solution are ensured. The role of the osmolality compensator in the diluent is to further ensure the concentration of the whole blood sample diluent, thereby maintaining the osmolality of the diluent. The buffer is used to adjust the pH of the diluent.
The invention has the beneficial effects that: the diluent for the blood analyzer improves the stability and the measurement reliability of a blood sample substance by compounding the stabilizer isopropanol and the 1, 2-propylene glycol in the formula, and can be suitable for the blood analyzers of different manufacturers by fixing the basic formula of the diluent and only adjusting the contents of the buffer and the osmotic pressure compensator, thereby simplifying the development steps of in-vitro diagnostic reagents and reducing the research and development cost. And breaks monopoly of reagents of original plants, and reduces the use cost of reagents of all levels of medical institutions.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below. It is to be understood that the described embodiments are merely a few embodiments of the invention, and not all embodiments.
The purity of the reagents used in the examples of the present invention was analytical grade.
The diluent comprises the following components: EDTA-K20.2-1.0 g/L of isopropanol, 0.5-1.5 g/L of isopropanol, 0.2-1.2 g/L of 1, 2-propylene glycol, 5.0-10.0 g/L, Tris-HCl, 0.6-1.5 g/L of sodium chloride and 0.3-0.5 g/L of potassium hydrogen phthalate.
The invention first determines EDTA-K2Isopropanol, 1, 2-propanediol and potassium hydrogen phthalate, and secondly adjusting the Tris-HCl content to enable the dilution to be analysed with different bloodThe pH value of the instrument is matched, and the amount of sodium chloride serving as an osmotic pressure compensator is adjusted according to the on-machine result of the adopted blood analyzer, so that the average red blood cell volume (MCV) detected on the on-machine instrument is consistent with that of the original reagent. Finally, the diluent is used for analyzing and detecting blood cells on a blood cell analyzer, and the applicability of the diluent to different models is evaluated according to the correlation between the diluent and the detection result of the original factory reagent. The test is carried out by taking ABXMICROS 60, ABX Pentra 80 and MEK-6318 models as examples, but is not limited to the three models. Meanwhile, in order to verify that the reliability and the stability of the detection data of the diluent are better than those of the diluent added with only one stabilizer by compounding the stabilizer isopropanol and the 1, 2-propylene glycol, the invention carries out the following embodiments.
The components were accurately weighed according to the following basic formulation in table 1, the PH of the solution was controlled by adding different amounts of Tris-HCl buffer, and the osmolality and conductivity of the solution were adjusted by adding sodium chloride to base solutions of different PH. After dissolving in sequence, using purified water to fix the volume, and filtering by using a 0.2 mu m microporous filter membrane to obtain the diluent for the blood analyzer. According to different PH and sodium chloride, the diluent formula suitable for different blood analyzers is determined.
TABLE 1
Example 1
A base solution having PH =7.1 was prepared according to the above table, and then 6.0g of sodium chloride was added to the base solution. The osmotic pressure of example 1 was 325 mOsm/kg; the conductivity was 18.9mS/cm.
Fresh blood of 5 healthy people is taken, the diluent of the invention in the example is used for testing on an ABX MICROROS 60 blood analyzer by using the original factory diluent and the invention diluent respectively, and blood cell analysis and detection results are compared. The results of the experiment are shown in table 2.
TABLE 2 comparison of the results of blood cell analysis and measurement of the reagent of the present invention on ABX MICROROS 60 blood analyzer with the results of the reagent of the original factory
Example 2
A base solution having PH =8.0 was prepared according to the above table, and 7.2g of sodium chloride was added to the base solution. The osmotic pressure of example 2 was 340 mOsm/kg; the conductivity was 18.2mS/cm.
Fresh blood of 5 healthy people is taken, the diluent of the invention of the second embodiment is used for testing on an ABX Pentra 80 blood analyzer by using the original factory diluent and the invention diluent respectively, and the analysis and detection results of blood cells are compared. The results of the experiment are shown in table 3.
TABLE 3 comparison of the result of blood cell analysis and detection of the reagent of the present invention in ABX Pentra 80 blood analyzer with the experimental result of the reagent of the original factory
Example 3
A base solution having PH =7.3 was prepared according to the above table, and then 7.0g of sodium chloride was added to the base solution. The osmotic pressure of example three was 330 mOsm/kg; the conductivity was 18.15mS/cm.
Fresh blood of 5 healthy people is taken, the diluent of the invention of the third embodiment is used for testing on an MEK-6318 blood analyzer by using the original factory diluent and the invention diluent respectively, and the differential counting result of the leucocytes is compared. The results of the experiment are shown in table 4.
TABLE 4 comparison of the results of blood cell analysis and detection of the reagent of the present invention in MEK-6318 blood analyzer with the results of the reagent experiment in the factory
Example 4
The diluent formulations are shown in table 5.
TABLE 5
The diluent A, B, C suitable for the three blood analyzers was prepared according to the above table, and the results of analyzing and detecting blood cells were compared with the reagents of the present invention of the first, second, and third examples. The comparative results are shown in Table 6.
Table 6 example 4 comparison of the results of the assay of the inventive reagents with the blood cells of example 1, example 2 and example 3 on a hematology analyzer.
As can be seen from the table, the pH of the solution is controlled within the range of 7.0-9.0, and the diluent prepared according to the invention can completely replace the original reagent to carry out blood cell analysis and detection on blood analyzers such as ABX MICROROS 60, ABX Pentra 80, MEK-6318 and the like. And by compounding the stabilizer isopropanol and the 1, 2-propylene glycol, the reliability and the stability of the detection data of the diluent are superior to those of the stabilizer only added with the isopropanol. In conclusion, the diluent for different blood analyzers provided by the invention has the advantages of simple operation, easy realization and lower cost of the prepared reagent, and can meet the requirements of most blood analyzers on analysis and detection of blood cells in the market.
Claims (8)
1. The diluent suitable for various blood analyzers is characterized by comprising a diluent basic formula, an osmotic pressure compensation agent 1.0-15.0 g/L and a buffering agent 0.6-1.5 g/L;
the basic formula of the diluent comprises 0.2-1.0 g/L anticoagulant, 0.3-0.5 g/L preservative and 0.2-1.5 g/L stabilizer;
the stabilizer is isopropanol and 1, 2-propylene glycol, and the concentration of the isopropanol is 0.1-2.0 g/L; the concentration of the 1, 2-propylene glycol is 0.1-1.5 g/L;
the anticoagulant is ethylenediamine tetraacetate, and the concentration of the anticoagulant is 0.2-1.0 g/L;
the osmotic pressure compensator is one or more of sodium chloride, potassium chloride, sodium sulfate and potassium sulfate.
2. The diluent of claim 1, wherein the anticoagulant is EDTA-K2。
3. The diluent of claim 1, wherein the preservative is one or more of sodium benzoate, potassium sorbate and potassium hydrogen phthalate, and the concentration is 0.3-0.5 g/L.
4. The diluent of claim 3, wherein the preservative is potassium hydrogen phthalate.
5. The diluent according to claim 1, wherein the concentration of the isopropanol is 0.5-1.5 g/L; the concentration of the 1, 2-propylene glycol is 0.2-1.2 g/L.
6. The diluent of claim 1, wherein the osmotic pressure compensator is sodium chloride; the concentration is 5.0-10.0 g/L.
7. The diluent of claim 1, wherein the buffer is one or more of a phosphate buffer, a Tris-HCl buffer, and a Good's buffer.
8. The diluent according to any one of claims 1 to 7, wherein the pH of the diluent is 7.0 to 9.0.
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| CN201711312375.1A CN108169468B (en) | 2017-12-12 | 2017-12-12 | Diluent suitable for various blood analyzers and preparation method thereof |
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| CN108169468B true CN108169468B (en) | 2020-04-07 |
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| CN113267395A (en) * | 2021-07-05 | 2021-08-17 | 江苏荣盛嘉美生物试剂有限公司 | Reagent for urine visible component analyzer and preparation method thereof |
| CN117783507B (en) * | 2024-02-27 | 2024-06-04 | 广州科方生物技术股份有限公司 | Component liquid commonly used in (1-3) -beta-D glucan and galactomannan detection, and preparation method and application thereof |
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| US6632676B1 (en) * | 1999-09-24 | 2003-10-14 | Clinical Diagnostic Solutions | Multi-purpose reagent system and method for enumeration of red blood cells, white blood cells and thrombocytes and differential determination of white blood cells |
| CN101078721B (en) * | 2006-05-23 | 2010-12-22 | 深圳迈瑞生物医疗电子股份有限公司 | Reagent and method for classifying leucocyte |
| CN101470108B (en) * | 2007-12-24 | 2013-11-27 | 深圳迈瑞生物医疗电子股份有限公司 | Reagent and method for classifying leukocyte |
| CN101819199B (en) * | 2010-05-08 | 2013-03-20 | 桂林优利特医疗电子有限公司 | Reagent for hemocyte analyzers |
| CN103743616A (en) * | 2013-12-19 | 2014-04-23 | 深圳市雷诺华科技实业有限公司 | Diluent for blood analyzer |
| CN104297134A (en) * | 2014-11-05 | 2015-01-21 | 深圳市开立科技有限公司 | Hemolytic agent and application thereof as well as classifying and counting method for white blood cells |
| CN104698157B (en) * | 2015-02-13 | 2017-05-17 | 中山市创艺生化工程有限公司 | Agent for blood cell analyzer |
| JP6518629B2 (en) * | 2015-07-06 | 2019-05-22 | 富士フイルム株式会社 | Blood test kit and blood analysis method |
| CN106885896A (en) * | 2017-03-30 | 2017-06-23 | 北京大文生物科技有限公司 | A kind of new hemolytic agent and preparation method thereof |
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