CN101464453A - Hemolytic agent - Google Patents
Hemolytic agent Download PDFInfo
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- CN101464453A CN101464453A CNA2007101252086A CN200710125208A CN101464453A CN 101464453 A CN101464453 A CN 101464453A CN A2007101252086 A CNA2007101252086 A CN A2007101252086A CN 200710125208 A CN200710125208 A CN 200710125208A CN 101464453 A CN101464453 A CN 101464453A
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- hemolytic agent
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Abstract
The invention discloses a hemolytic agent which contains at least one organic acid whose content ranges from 1 to 20g/L, and relates to a multipurpose hemolytic agent, which can be used for automatic hemanalysis, and can accurately test hematoglobin and keep the normal classification of leucocyte at the same time. A stabilizing agent of hematoglobin derivants is added in the hemolytic agent, thereby preventing the derivants from changing along with the reaction temperature change, and ensuring that an accurate and stable test result can be obtained within the normal service temperature range of the instrument.
Description
Technical field
The present invention relates to a kind of blood cell analyrical reagent, particularly relate to a kind of hemolytic agent that leukocyte differential count detects and content of hemoglobin is measured that can be used for.
Background technology
Hemolytic agent is one of the most frequently used reagent of blood cell analyzer, also is the key reagents of leucocyte and respectively hive off counting, HGB assay.After diluted blood adds hemolytic agent, erythrocytolysis, discharge haemoglobin, relevant composition is in conjunction with forming haemoglobin dervative in the latter and the hemolytic agent, enter examples of hemoglobin detection system, (generally 530～550nm) times colorimetrics, the variation of absorbance is directly proportional with content of hemoglobin in the liquid, and instrument just can show its concentration at specific wavelength; Simultaneously, dissimilar leucocyte (lymphocyte, monocyte, neutrophil leucocyte etc.) forms evident difference after the hemolytic agent effect, according to these difference, instrument can be divided into the leucocyte in the blood sample several monoids, and the quantity of total white blood cells and each monoid is added up.
1996, ICSH recommended cyanmethemoglobin (HiCN) determination method as international HGB bioassay standard method, and its Main physical index is: at crest λ=540nm, during trough λ=504nm, have tangible crest, trough.Though this method can more accurately be measured HGB, can not measure WBC simultaneously, can't be applied to the clinical detection instrument.In addition, KCN has severe toxicity, not only serious harm laboratory worker healthy, and environmental pollution is also very serious simultaneously.
At present, no cyaniding hemolytic agent commonly used substitutes the KCN use in the clinical examination both at home and abroad, and existing correlation technique is as follows:
Prior art one: mainly use cetyl trimethyl ammonium bromide, Triton X-100, medical grade second alcohol and water to form cyanogens-free hemolytic agent;
Prior art two: mainly use organic quaternary ammonium salt, oxammonium hydrochloride, dipotassium hydrogen phosphate to form cyanogens-free hemolytic agent;
Prior art three: mainly use cationic surfactant to be used for lysed erythrocyte and make the haemoglobin sex change, use simultaneously to comprise sulfosalicylic acid, nitrogenous and the water-soluble chelator of carboxyl and at least a stabilized hemoglobin agent of piperazine;
Prior art four: mainly use at least a lysed erythrocyte that is used in quaternary ammonium salt, the pyridiniujm, discharge haemoglobin; Use a kind of antioxidant simultaneously, be selected from the oxyacid alkali metal salt of phosphoric acid, sodium sulphite, sodium pyrosulfite, sodium bisulfite, sodium thiosulfate, other sulphur.
Above prior art is paid close attention to the accuracy of hemoglobinometry under the normal temperature condition mostly, can not guarantee to take place still to obtain stable and accurate measurement result after the bigger variation in the serviceability temperature scope.
Summary of the invention
The objective of the invention is at the deficiencies in the prior art, a kind of accuracy and conforming hemolytic agent that can still guarantee the content of hemoglobin measurement result when temperature takes place by bigger the variation is provided.
For achieving the above object, the present invention has adopted following technical scheme:
The invention discloses a kind of hemolytic agent, described hemolytic agent contains at least a organic acid, and described organic acid content is 1～20g/L.
In the specific embodiment of the present invention, described organic acid is preferably ascorbic acid or sulfanilic acid.
Hemolytic agent of the present invention preferably contain at least a can lysed erythrocyte and discharge the surfactant of haemoglobin.
Described surfactant is preferably cationic surfactant, more preferably quaternary cationic surfactant.
Further, described surfactant is preferably DTAC, DTAB, hexadecyltrimethylammonium chloride or cetyl trimethyl ammonium bromide.
In the concrete embodiment of the present invention, the content of described surfactant in described hemolytic agent is preferably 2～4%, and described is weight percentage.
Further contain buffering agent in the hemolytic agent of the present invention, the pH that described buffering agent is regulated hemolytic agent is 5～9.
Wherein, described buffering agent is preferably phosphate buffer.
Owing to adopted above technical scheme, the beneficial effect that the present invention is possessed is:
Hemolytic agent of the present invention can be used for measuring parameters such as total white blood cells, each differential count and content of hemoglobin with after blood sample mixes.Wherein, formed more stable haemoglobin compound owing to added the compound organic acid that combines with the haemoglobin dervative specificity.Bigger variation has taken place in serviceability temperature in this compound except keeping stablizing under the normal normal temperature condition that uses, still keep stable in the usable range more widely as 10～40 ℃.Reagent of the present invention all can keep the temperature effect of haemoglobin test result very little in 10～40 ℃ of temperature ranges, and relative deviation remains in 3% between 25 ℃ of 10 ℃ and 40 ℃ and the normal temperature.Changed that the compound that haemoglobin forms varies with temperature, the present situation of significant change takes place its light absorption value thereupon under common hemolytic agent effect, thus when having guaranteed that content of hemoglobin is measured the apparatus measures result accurately and consistance.Widened the Applicable temperature scope for clinical use, reduced when using apparatus measures, made general medium and small check section office when the stationary temperature condition can not be provided comparatively, still can obtain testing result accurately the requirement of laboratory environment temperature.
Hemolytic agent of the present invention can also be used for leukocytic normal classification and Detection, when lysed erythrocyte discharges haemoglobin, leucocyte is modified, make each monoid of leucocyte produce volume difference, realize leukocytic three hive off detections and the accuracys of assurance testing result.
Embodiment
The present invention relates to a kind of multi-usage hemolytic agent, can be used for automated blood analysis, accurate hemoglobin testing when keeping the normal classification of leucocyte.Wherein emphasis solves the problem that the haemoglobin detected value changes with temperature.Added the stabilizing agent of haemoglobin dervative in the hemolytic agent of the present invention, avoided derivant to change, guaranteed in the normal serviceability temperature scope of instrument, all can obtain accurate, stable test result with the variation of temperature of reaction.
Hemolytic agent among the present invention is mainly used in lysed erythrocyte, keeps all kinds of characters of leucocyte, and combines with the haemoglobin specificity, forms stable compound, and formation specificity absorption peak about 540nm obtains the HGB measured value according to the variation of absorbance.
Hemolytic agent among the present invention contains cationic surfactant, be mainly used in lysed erythrocyte, discharge haemoglobin, and pass through leukocytic modification, make each monoid of leucocyte produce volume difference, thereby can realize that leucocyte three hives off by electrical impedance method, promptly be divided into lymphocyte (Lymph), osculant cell (monocyte, acidophil, basocyte, Mid) and three monoids of neutrophil leucocyte (Gran).In the present invention, this area commonly used, solubilized red blood cell and to discharge the cationic surfactant of haemoglobin all applicable.Wherein, preferably use quaternary cationic surfactant, its addition is about 2～4%, and described is weight percentage, as: DTAC, DTAB etc.
Hemolytic agent among the present invention contains the special component organic acid, be used for combining, form stabilized complex, when ambient temperature changes with haemoglobin dervative, still can keep the result of haemoglobin compound stable, thereby obtain accurate, stable HGB measurement result.
The preferred following two kinds of organic acids of organic acid among the present invention---
Ascorbic acid (Ascorbic Acid), structural formula is as follows:
Sulfanilic acid (p-aminobenzene sulfonic acid), structural formula is as follows:
Through experimental verification, described organic acid can combine with the haemoglobin dervative specificity, plays function of stabilizer, thereby it is stable to guarantee that haemoglobin dervative all can be kept morphological feature in 10～40 ℃ scope, light absorption value does not have marked change, promptly guarantees the stable of HGB measured value.
Among the present invention, also contain and adjust the buffering agent that PH uses.Do not have specific requirement for buffering agent, can preferably use phosphate buffer for using buffer system such as formic acid, phthalic acid, acetate, phosphoric acid, TRIS, boric acid, carbonic acid etc. always.The PH scope does not have absolute effect to classification among the present invention, adjusts to usually in 5～9 scopes.Use amount is 10～200mM.
The present invention can provide WBC and three accurate countings that hive off accurately through clinical verification, and stable, real HGB measurement result, guarantees that simultaneously measurement result does not change with temperature fluctuation, has guaranteed that between 10～40 degree HGB result's is reliable and stable.
The present invention realizes erythrocytic quick dissolving by the cationic surfactant in the hemolytic agent, discharges haemoglobin; Combine and form stabilized complex by organic acid with the haemoglobin specificity, make the effect of hemolytic agent not be subjected to the influence of variation of ambient temperature; Guarantee that by organic buffer agent and/or inorganic buffer agent the dilution system has stable p H value environment.Wherein, use organic acid as the stabilized hemoglobin agent first, can change the compound that haemoglobin forms and vary with temperature under common hemolytic agent effect, the present situation of significant change takes place in its light absorption value thereupon.Widened the Applicable temperature scope for clinical use, reduced when using apparatus measures, made general medium and small check section office when the stationary temperature condition can not be provided comparatively, still can obtain testing result accurately the requirement of laboratory environment temperature.
Below by specific embodiment the present invention is done further detailed description.
Embodiment 1
A kind of hemolytic agent has following composition:
DTAC 40g
Ascorbic acid 10g
Phosphate buffered solution 0.05M
Moisturizing is to 1L
pH 5.6
Use Mai Rui company to produce BC-3000Plus liquid cytoanalyze, the supporting dilution that uses manufacturer to provide, and the hemolytic agent of the above prescription of use are analyzed automatically.Detailed process is carried out according to the method that manufacturer provides, and specifically comprises: whole blood and dilution mix, and form sample after the dilution of first concentration.With sample separated into two parts behind first concentration dilution.Sample mixes with dilution after getting part first concentration dilution, forms the sample of second concentration, is used for red blood cell and enumeration of thrombocytes and measures.Sample mixes with hemolytic agent behind the first remaining concentration dilution, forms the sample of the 3rd concentration, is used for leukocytic count measurement and hemoglobin concentration.
Measure by above-mentioned steps respectively under 10 ℃, 25 ℃ and 40 ℃ conditions of room temperature, each temperature conditions is chosen six different fresh blood samples and is tested, and the results are shown in shown in the following table 1.
Table 1
The result: 10 ℃ of HGB deviations than 25 ℃ are 0.37%;
40 ℃ of HGB deviations than 25 ℃ are 2.8%
Embodiment 2
A kind of hemolytic agent has following composition:
DTAB 40g
Sulfanilic acid 10g
Phosphate buffered solution 0.05M
Moisturizing is to 1L
pH 7.6
The hemolytic agent of the present embodiment of blood sampling, and use in addition, all the other are measured by embodiment 1 described method, and the result is as shown in table 2.
Table 2
The result: 10 ℃ of HGB deviations than 25 ℃ are 0.36%;
40 ℃ of HGB deviations than 25 ℃ are 2.7%
Embodiment 3
A kind of hemolytic agent has following composition:
Hexadecyltrimethylammonium chloride 20g
Ascorbic acid 1g
Phosphate buffered solution 0.05M
Moisturizing is to 1L
pH 5.6
The hemolytic agent of the present embodiment of blood sampling, and use in addition, all the other are measured by embodiment 1 described method, and the result is as shown in table 3.
Table 3
The result: 10 ℃ of HGB deviations than 25 ℃ are 1.9%;
40 ℃ of HGB deviations than 25 ℃ are 2.6%
Embodiment 4
A kind of hemolytic agent has following composition:
Cetyl trimethyl ammonium bromide 20g
Sulfanilic acid 20g
Phosphate buffered solution 0.05M
Moisturizing is to 1L
pH 7.6
The hemolytic agent of the present embodiment of blood sampling, and use in addition, all the other are measured by embodiment 1 described method, and the result is as shown in table 4.
Table 4
The result: 10 ℃ of HGB deviations than 25 ℃ are 1.5%;
40 ℃ of HGB deviations than 25 ℃ are 2.9%
Above content be in conjunction with concrete preferred implementation to further describing that the present invention did, can not assert that concrete enforcement of the present invention is confined to these explanations.For the general technical staff of the technical field of the invention, without departing from the inventive concept of the premise, can also make some simple deduction or replace, all should be considered as belonging to protection scope of the present invention.
Claims (9)
1. hemolytic agent, it is characterized in that: described hemolytic agent contains at least a organic acid, and described organic acid content is 1～20g/L.
2, hemolytic agent according to claim 1 is characterized in that: described organic acid is ascorbic acid or sulfanilic acid.
3, hemolytic agent according to claim 1 and 2 is characterized in that: described hemolytic agent contain at least a can lysed erythrocyte and discharge the surfactant of haemoglobin.
4, hemolytic agent according to claim 3 is characterized in that: described surfactant is a cationic surfactant.
5, hemolytic agent according to claim 4 is characterized in that: described surfactant is a quaternary cationic surfactant.
6, hemolytic agent according to claim 5 is characterized in that: described surfactant is DTAC, DTAB, hexadecyltrimethylammonium chloride or cetyl trimethyl ammonium bromide.
7, according to any described hemolytic agent of claim 4～6, it is characterized in that: the content of described surfactant in described hemolytic agent is 2～4%, and described is weight percentage.
8, hemolytic agent according to claim 3 is characterized in that: described hemolytic agent contains buffering agent, and the pH that this buffering agent is regulated hemolytic agent is 5～9.
9, hemolytic agent according to claim 8 is characterized in that: described buffering agent is a phosphate buffer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN200710125208.6A CN101464453B (en) | 2007-12-18 | 2007-12-18 | A kind of hemolytic agent |
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| CN200710125208.6A CN101464453B (en) | 2007-12-18 | 2007-12-18 | A kind of hemolytic agent |
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| CN101464453A true CN101464453A (en) | 2009-06-24 |
| CN101464453B CN101464453B (en) | 2016-09-07 |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102600918A (en) * | 2012-03-09 | 2012-07-25 | 深圳联合医学科技有限公司 | Microfluidic chip for blood analysis and method for using microfluidic chip |
| CN103323581A (en) * | 2013-06-18 | 2013-09-25 | 南京普朗医疗设备有限公司 | Hemocyte analyzer hemolytic agent |
| CN106932350A (en) * | 2017-03-22 | 2017-07-07 | 安徽民泰医药科技有限公司 | Blood cell analysis hemolytic agent |
| CN107144519A (en) * | 2017-05-24 | 2017-09-08 | 中山市滔略生物科技有限公司 | Cellanalyzer hemolytic agent and preparation method thereof |
| CN108318408A (en) * | 2018-01-30 | 2018-07-24 | 深圳唯公生物科技有限公司 | The pretreating reagent and method of leukocyte differential count sample |
| CN110687306A (en) * | 2019-10-30 | 2020-01-14 | 深圳上泰生物工程有限公司 | Direct-connection on-board hemolytic enzyme method double-reagent glycosylated hemoglobin detection kit |
| CN113218847A (en) * | 2021-04-12 | 2021-08-06 | 武汉凯普瑞生物技术有限公司 | Hemolytic agent for flow cytometry analysis and preparation method and application method thereof |
| CN114279777A (en) * | 2020-09-28 | 2022-04-05 | 深圳市瑞图生物技术有限公司 | Hemolytic agent containing amidino compound and blood analysis kit thereof |
Citations (1)
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| CN1490622A (en) * | 2003-08-20 | 2004-04-21 | 深圳迈瑞生物医疗电子股份有限公司 | Cyanideless hemolysin and its use |
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Patent Citations (1)
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| CN1490622A (en) * | 2003-08-20 | 2004-04-21 | 深圳迈瑞生物医疗电子股份有限公司 | Cyanideless hemolysin and its use |
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| 夏丽: "高浓度抗坏血酸对人离体红细胞的毒性和临床意义", 《中国优秀博硕士学位论文全文数据库（硕士）医药卫生科技辑》 * |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102600918A (en) * | 2012-03-09 | 2012-07-25 | 深圳联合医学科技有限公司 | Microfluidic chip for blood analysis and method for using microfluidic chip |
| CN102600918B (en) * | 2012-03-09 | 2014-06-18 | 深圳联合医学科技有限公司 | Microfluidic chip for blood analysis and method for using microfluidic chip |
| CN103323581A (en) * | 2013-06-18 | 2013-09-25 | 南京普朗医疗设备有限公司 | Hemocyte analyzer hemolytic agent |
| CN106932350A (en) * | 2017-03-22 | 2017-07-07 | 安徽民泰医药科技有限公司 | Blood cell analysis hemolytic agent |
| CN107144519A (en) * | 2017-05-24 | 2017-09-08 | 中山市滔略生物科技有限公司 | Cellanalyzer hemolytic agent and preparation method thereof |
| CN108318408A (en) * | 2018-01-30 | 2018-07-24 | 深圳唯公生物科技有限公司 | The pretreating reagent and method of leukocyte differential count sample |
| CN108318408B (en) * | 2018-01-30 | 2020-04-17 | 深圳唯公生物科技有限公司 | Reagent and method for pretreating sample for classifying leukocytes |
| CN110687306A (en) * | 2019-10-30 | 2020-01-14 | 深圳上泰生物工程有限公司 | Direct-connection on-board hemolytic enzyme method double-reagent glycosylated hemoglobin detection kit |
| CN110687306B (en) * | 2019-10-30 | 2023-04-25 | 深圳上泰生物工程有限公司 | Double-reagent glycosylated hemoglobin detection kit for direct on-machine hemolysis enzyme method |
| CN114279777A (en) * | 2020-09-28 | 2022-04-05 | 深圳市瑞图生物技术有限公司 | Hemolytic agent containing amidino compound and blood analysis kit thereof |
| CN113218847A (en) * | 2021-04-12 | 2021-08-06 | 武汉凯普瑞生物技术有限公司 | Hemolytic agent for flow cytometry analysis and preparation method and application method thereof |
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| Publication number | Publication date |
|---|---|
| CN101464453B (en) | 2016-09-07 |
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Effective date of registration: 20180808 Address after: 518057 the 1-4 floor of MINDRAY building, science and technology south twelve Road, Nanshan District high tech Industrial Park, Shenzhen, Guangdong. Co-patentee after: Shenzhen MINDRAY Technology Co., Ltd. Patentee after: Shenzhen Mairui Biotherapeutic Electronic Co., Ltd. Address before: 518057 MINDRAY science and technology south twelve road MINDRAY high tech Industrial Park, Shenzhen, Guangdong Patentee before: Shenzhen Mairui Biotherapeutic Electronic Co., Ltd. |