CN101464453B - A kind of hemolytic agent - Google Patents
A kind of hemolytic agent Download PDFInfo
- Publication number
- CN101464453B CN101464453B CN200710125208.6A CN200710125208A CN101464453B CN 101464453 B CN101464453 B CN 101464453B CN 200710125208 A CN200710125208 A CN 200710125208A CN 101464453 B CN101464453 B CN 101464453B
- Authority
- CN
- China
- Prior art keywords
- hemolytic agent
- agent
- hemolytic
- surfactant
- organic acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a kind of hemolytic agent, described hemolytic agent contains at least one organic acid, and the content of described organic acid is 1～20g/L.The present invention relates to a kind of multipurpose hemolytic agent, can be used for automated blood analysis, can accurate hemoglobin testing while keeping leukocyte normally to classify, hemolytic agent of the present invention with the addition of the stabilizer of haemoglobin dervative, derivant is avoided to change with the change of reaction temperature, it is ensured that to be all obtained in that accurate, stable test result in instrument normally uses temperature range.
Description
Technical field
The present invention relates to a kind of blood cell analysis reagent, especially relate to one and can be used for leukocyte differential count detection
And the hemolytic agent that content of hemoglobin measures.
Background technology
Hemolytic agent is one of the most frequently used reagent of blood cell analyzer, is also leukocyte and the counting that respectively hives off, HGB content
The key reagents measured.After the blood being diluted adds hemolytic agent, erythrocytolysis, discharge hemoglobin, the latter and hemolytic agent
Middle Related Component combines and forms haemoglobin dervative, enters examples of hemoglobin detection system, specific wavelength (typically 530～
Colorimetric under 550nm), the change of absorbance is directly proportional to content of hemoglobin in liquid, and instrument just can show its concentration；Meanwhile,
Different types of leukocyte (lymphocyte, mononuclear cell, neutrophilic granulocyte etc.) forms obvious difference after hemolytic agent effect,
According to these difference, the leukocyte in blood sample can be divided into several monoid by instrument, and to total white blood cells and each monoid
Quantity add up.
1996, ICSH recommended cyanmethemoglobin (HiCN) algoscopy as the world
HGB bioassay standard method, its main physical indexes is: at crest λ=540nm, during trough λ=504nm, have obvious ripple
Peak, trough.Although this method can relatively accurately measure HGB, but can not measure WBC, it is impossible to is applied to Clinical detection instrument simultaneously.
Additionally, KCN has severe toxicity, not only serious harm laboratory worker healthy, environmental pollution is the most serious simultaneously.
At present, conventional without the replacement KCN use of cyaniding hemolytic agent in domestic and international Clinical Laboratory, existing correlation technique is as follows:
Prior art one: mainly use cetyl trimethylammonium bromide, Triton X-100, medical grade ethanol
Cyanogens-free hemolytic agent is formed with water；
Prior art two: mainly use organic quaternary ammonium salt, oxammonium hydrochloride., dipotassium hydrogen phosphate to form cyanogens-free hemolytic agent；
Prior art three: main use cationic surfactant for lysed erythrocyte and makes hemoglobin degeneration, with
Time use at least one hemoglobin including sulfosalicylic acid, the nitrogenous and water-soluble chelator of carboxyl and piperazine steady
Determine agent；
Prior art four: mainly use at least one in quaternary ammonium salt, pyridiniujm for lysed erythrocyte, discharge blood red egg
In vain；Use a kind of antioxidant simultaneously, selected from phosphoric acid, sodium sulfite, sodium pyrosulfite, sodium bisulfite, sodium thiosulfate, its
The oxyacid alkali metal salt of his sulfur.
Above prior art pays close attention to the accuracy of hemoglobinometry under normal temperature condition mostly, it is impossible to ensure using temperature
Stable and accurate measurement result is still obtained after scope generation large change.
Summary of the invention
It is an object of the invention to for the deficiencies in the prior art, it is provided that one can when temperature generation large change still
Ensure the accuracy of content of hemoglobin measurement result and conforming hemolytic agent.
For achieving the above object, present invention employs techniques below scheme:
The invention discloses a kind of hemolytic agent, described hemolytic agent contains at least one organic acid, and described organic acid
Content is 1～20g/L.
In the detailed description of the invention of the present invention, described organic acid is preferably ascorbic acid or p-aminobenzene sulfonic acid.
The hemolytic agent of the present invention preferably comprises at least one can be with lysed erythrocyte the surface activity discharging hemoglobin
Agent.
Described surfactant is preferably cationic surfactant, more preferably quaternaries cation surface active
Agent.
Further, described surfactant is preferably Dodecyl trimethyl ammonium chloride, trimethyl bromination
Ammonium, hexadecyltrimethylammonium chloride or cetyl trimethylammonium bromide.
In specific embodiment of the present invention, described surfactant content in described hemolytic agent be preferably 2～
4%, described for weight percentage.
Containing buffer agent in the hemolytic agent of the present invention further, the pH of described buffer agent regulation hemolytic agent is 5～9.
Wherein, described buffer agent is preferably phosphate buffer.
Owing to have employed above technical scheme, make what the present invention possessed to have the beneficial effects that:
After the hemolytic agent of the present invention mixes with blood sample, can be used for measuring total white blood cells, each differential counting and hemoglobin
The parameters such as content.Wherein owing to the addition of the compound organic acid specific binding with haemoglobin dervative, define more
Stable haemoglobin complexes.This complex, in addition to keeping stablizing under the normal normal temperature condition used, is using temperature
There occurs large change, as stable in remained in that in 10～40 DEG C more widely range.The reagent of the present invention 10～
The temperature effects that all can keep hemoglobin test result in 40 DEG C of temperature ranges is the least, between 10 DEG C and 40 DEG C and room temperature 25 DEG C
Within relative deviation is maintained at 3%.Change the complex that hemoglobin formed under common hemolytic agent effect vary with temperature,
There is the present situation of significant change in its light absorption value, thus ensure that the accurate of apparatus measures result when content of hemoglobin measures therewith
And concordance.Widen Applicable temperature scope for Clinical practice, reduced when using apparatus measures laboratory ambient temperature
Requirement so that general medium and small inspection section office still are able to obtain when not being provided that more stationary temperature condition detects knot accurately
Really.
Hemolytic agent of the present invention can be used for the normal classification and Detection of leukocyte, discharges the same of hemoglobin at lysed erythrocyte
Time leukocyte is modified, make each monoid of leukocyte produce volume difference, it is achieved the three of leukocyte hive off detection, and ensure to examine
Survey the accuracy of result.
Detailed description of the invention
The present invention relates to a kind of multipurpose hemolytic agent, can be used for automated blood analysis, normally classify keeping leukocyte
Simultaneously can accurate hemoglobin testing.Wherein emphasis solves the problem that hemoglobin detected value changes with temperature.This
Bright hemolytic agent with the addition of the stabilizer of haemoglobin dervative, it is to avoid derivant changes with the change of reaction temperature,
Ensure all to be obtained in that accurate, stable test result in instrument normally uses temperature range.
Hemolytic agent in the present invention is mainly used in lysed erythrocyte, keeps all kinds of character of leukocyte, and and hemoglobin
Specific binding, form stable compound, form specificity absworption peak at about 540nm, obtain according to the change of absorbance
HGB measured value.
Hemolytic agent in the present invention contains cationic surfactant, is mainly used in lysed erythrocyte, release hemoglobin,
And by the modification to leukocyte, make each monoid of leukocyte produce volume difference, such that it is able to realized white thin by electrical impedance method
Born of the same parents three hive off, be i.e. divided into lymphocyte (Lymph), Mononuclear cell (mononuclear cell, acidophil, basophilic leukocyte, Mid) and
Three monoids of neutrophilic granulocyte (Gran).In the present invention, commonly used in the art, solubilized erythrocyte discharge hemoglobin
Cationic surfactant is the most applicable.Wherein, quaternary cationic surfactant is preferably used, its addition 2～
About 4%, described for weight percentage, such as: Dodecyl trimethyl ammonium chloride, Dodecyl trimethyl ammonium chloride etc..
Hemolytic agent in the present invention contains special component organic acid, for being combined with haemoglobin dervative, is formed stable
Complex, when ambient temperature changes, the result being maintained to haemoglobin complexes is stable, thus obtains standard
Really, stable HGB measurement result.
Organic acid preferred following two organic acid in the present invention
Ascorbic acid (Ascorbic Acid), structural formula is as follows:
P-aminobenzene sulfonic acid (p-aminobenzene sulfonic acid), structural formula is as follows:
Experiments verify that, described organic acid can be specific binding with haemoglobin dervative, plays the effect of stabilizer,
Thus ensureing that haemoglobin dervative is all able to maintain that in the range of 10～40 DEG C morphological characteristic is stable, light absorption value becomes without notable
Change, i.e. ensure stablizing of HGB measured value.
In the present invention, possibly together with the buffer agent adjusting PH.Requirement not specific for buffer agent, can be conventional slow
Rush system such as formic acid, phthalic acid, acetic acid, phosphoric acid, TRIS, boric acid, carbonic acid etc., phosphate buffer is preferably used.This
Bright middle PH scope does not has absolute effect to classification, in the range of being generally adjusted to 5～9.Usage amount is 10～200mM.
The present invention is according to clinical verification, it is provided that WBC and three accurate countings hived off accurately, and stable, true
HGB measurement result, simultaneously ensure measurement result do not change with temperature fluctuation, it is ensured that between 10～40 degree HGB tie
Really reliable and stable.
The present invention realizes erythrocytic rapid solution by the cationic surfactant in hemolytic agent, discharges blood red egg
In vain；Specific binding by organic acid and hemoglobin and form stable complex so that the effect of hemolytic agent is not by environment
The impact of variations in temperature；Ensure that diluent system has stable pH environment by organic buffer agent and/or inorganic buffer agent.Its
In, use organic acid as stabilized hemoglobin agent first, thus it is possible to vary hemoglobin is formed under common hemolytic agent effect
Complex varies with temperature, and its light absorption value occurs the present situation of significant change therewith.Applicable temperature scope has been widened for Clinical practice,
Reduce the requirement to laboratory ambient temperature when using apparatus measures so that general medium and small inspection section office be not provided that more
It still is able to obtain testing result accurately during stationary temperature condition.
Below by specific embodiment, the present invention is done further detailed description.
Embodiment 1
A kind of hemolytic agent, has a consisting of:
Dodecyl trimethyl ammonium chloride 40g
Ascorbic acid 10g
Phosphate buffered solution 0.05M
Moisturizing is to 1L
pH 5.6
Use MINDRAY to produce BC-3000 Plus blood cell analyzer, use the supporting diluent that manufacturer provides,
And use the hemolytic agent of above formula, automatically analyze.The method that detailed process provides according to manufacturer is carried out, and specifically includes:
Whole blood and diluent mixing, form sample after the dilution of the first concentration.Sample after first concentration dilution is divided into two parts.Take portion
After dividing the first concentration dilution, sample mixes with diluent, forms the sample of the second concentration, for erythrocyte and enumeration of thrombocytes
Measure.After remaining first concentration dilution, sample mixes with hemolytic agent, forms the sample of the 3rd concentration, for the counting of leukocyte
Measure and hemoglobin concentration.
Respectively at 10 DEG C, be measured by above-mentioned steps under the conditions of room temperature 25 DEG C and 40 DEG C, each temperature conditions chooses six
The fresh blood sample of Zhi Butong is tested, and result see table shown in 1.
Table 1
Result: the HGB deviation of 10 DEG C relatively 25 DEG C is 0.37%；
The HGB deviation of 40 DEG C relatively 25 DEG C is 2.8%
Embodiment 2
A kind of hemolytic agent, has a consisting of:
Dodecyl trimethyl ammonium chloride 40g
P-aminobenzene sulfonic acid 10g
Phosphate buffered solution 0.05M
Moisturizing is to 1L
pH 7.6
Another blood sampling, and use the hemolytic agent of the present embodiment, remaining method as described in embodiment 1 is measured, result
As shown in table 2.
Table 2
Result: the HGB deviation of 10 DEG C relatively 25 DEG C is 0.36%；
The HGB deviation of 40 DEG C relatively 25 DEG C is 2.7%
Embodiment 3
A kind of hemolytic agent, has a consisting of:
Hexadecyltrimethylammonium chloride 20g
Ascorbic acid 1g
Phosphate buffered solution 0.05M
Moisturizing is to 1L
pH 5.6
Another blood sampling, and use the hemolytic agent of the present embodiment, remaining method as described in embodiment 1 is measured, result
As shown in table 3.
Table 3
Result: the HGB deviation of 10 DEG C relatively 25 DEG C is 1.9%；
The HGB deviation of 40 DEG C relatively 25 DEG C is 2.6%
Embodiment 4
A kind of hemolytic agent, has a consisting of:
Cetyl trimethylammonium bromide 20g
P-aminobenzene sulfonic acid 20g
Phosphate buffered solution 0.05M
Moisturizing is to 1L
pH 7.6
Another blood sampling, and use the hemolytic agent of the present embodiment, remaining method as described in embodiment 1 is measured, result
As shown in table 4.
Table 4
Result: the HGB deviation of 10 DEG C relatively 25 DEG C is 1.5%；
The HGB deviation of 40 DEG C relatively 25 DEG C is 2.9%
Above content is to combine concrete preferred implementation further description made for the present invention, it is impossible to assert
Being embodied as of the present invention is confined to these explanations.For general technical staff of the technical field of the invention,
On the premise of present inventive concept, it is also possible to make some simple deduction or replace, all should be considered as belonging to the present invention's
Protection domain.
Claims (8)
1. a multipurpose hemolytic agent, it is characterised in that: described hemolytic agent contains at least one stabilizing hemoglobin derivant
Organic acid, and the content of described organic acid is 1-20g/L；Described hemolytic agent contains at least one can be with lysed erythrocyte and release
The surfactant of blood-letting Lactoferrin, described hemolytic agent is for hemoglobin concentration or leukocyte differential count detection.
Hemolytic agent the most according to claim 1, it is characterised in that: described organic acid is ascorbic acid or p-aminophenyl sulphur
Acid.
Hemolytic agent the most according to claim 1, it is characterised in that: described surfactant is cationic surfactant.
Hemolytic agent the most according to claim 3, it is characterised in that: described surfactant is quaternaries cation surface
Activating agent.
Hemolytic agent the most according to claim 4, it is characterised in that: described surfactant is trimethyl chlorination
Ammonium, Dodecyl trimethyl ammonium chloride, hexadecyltrimethylammonium chloride or cetyl trimethylammonium bromide.
6. according to the hemolytic agent described in claim 1-5 any one, it is characterised in that: described surfactant is at described haemolysis
Content in agent is 2-4%, and described content is weight percentage.
Hemolytic agent the most according to claim 1, it is characterised in that: described hemolytic agent contains buffer agent, and this buffer agent regulates
The pH of hemolytic agent is 5-9.
Hemolytic agent the most according to claim 7, it is characterised in that: described buffer agent is phosphate buffer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200710125208.6A CN101464453B (en) | 2007-12-18 | 2007-12-18 | A kind of hemolytic agent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200710125208.6A CN101464453B (en) | 2007-12-18 | 2007-12-18 | A kind of hemolytic agent |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101464453A CN101464453A (en) | 2009-06-24 |
| CN101464453B true CN101464453B (en) | 2016-09-07 |
Family
ID=40805131
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200710125208.6A Active CN101464453B (en) | 2007-12-18 | 2007-12-18 | A kind of hemolytic agent |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101464453B (en) |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| 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 |
| 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 |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
-
2007
- 2007-12-18 CN CN200710125208.6A patent/CN101464453B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
Non-Patent Citations (1)
| Title |
|---|
| 李树德等.抗坏血酸增强氯高铁血红素诱导的溶血.《中国生物化学与分子生物学报》.2006,第22卷(第2期),158-162. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101464453A (en) | 2009-06-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101464453B (en) | A kind of hemolytic agent | |
| EP1298441B1 (en) | A cyanide-free lytic reagent composition and method for hemoglobin and cell analysis | |
| US7235404B2 (en) | Cyanide-free lytic reagent composition and method of use for hemoglobin and white blood cell measurement | |
| US5882934A (en) | Composition and method for hemoglobin and cell analysis | |
| CN103698501B (en) | A kind of Cyanide-free hemolytic agent | |
| Rodkey | Direct spectrophotometric determination of albumin in human serum | |
| Roberts et al. | Red blood cell distribution width index in some hematologic diseases | |
| Piva et al. | Automated reticulocyte counting: state of the art and clinical applications in the evaluation of erythropoiesis | |
| JP4153162B2 (en) | Blood diluent | |
| WO1998032016A9 (en) | A cyanide-free lytic reagent composition and method for hemoglobin and cell analysis | |
| US6632676B1 (en) | Multi-purpose reagent system and method for enumeration of red blood cells, white blood cells and thrombocytes and differential determination of white blood cells | |
| CN101451931B (en) | Blood dilution liquid and use method thereof | |
| EP2324350B1 (en) | Reference control for cell by cell analysis | |
| JPH10503016A (en) | Cyanide-free assay method and reagents for hemoglobin and leukocytes in whole blood | |
| CN101329229A (en) | Cyanogen-free leucocyte tri-grouping environment protection type haemolysin for blood cell analysis | |
| US11835532B2 (en) | Green concentrated reagent for hemotology systems | |
| US6890756B2 (en) | Method of using cyanide-free lyse solution to emulate a cyanide-containing lyse solution in the measurement of hemoglobin | |
| CN108169468B (en) | Diluent suitable for various blood analyzers and preparation method thereof | |
| Gross et al. | Glutamate-induced efflux of protein, neuron-specific enolase and lactate dehydrogenase from a mesencephalic cell culture | |
| CN110160941A (en) | Three classification reagent for hemocyte analyzers of one kind | |
| CN114047107A (en) | Diluent for blood cell analysis | |
| Anderson et al. | Dynamic NMR measurement of volume regulatory changes in Amphiuma RBC Na+ content | |
| CN110954489A (en) | Cyanide-free hemolytic agent and application thereof | |
| Iosefsohn et al. | Ektachem multilayer dry-film assay for ammonia evaluated. |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
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. |
|
| TR01 | Transfer of patent right |