CN106932350A - Blood cell analysis hemolytic agent - Google Patents
Blood cell analysis hemolytic agent Download PDFInfo
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- CN106932350A CN106932350A CN201710174670.9A CN201710174670A CN106932350A CN 106932350 A CN106932350 A CN 106932350A CN 201710174670 A CN201710174670 A CN 201710174670A CN 106932350 A CN106932350 A CN 106932350A
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- hemolytic agent
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- blood cell
- cell analysis
- pyrithione
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
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- 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
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Abstract
The present invention proposes a kind of blood cell analysis hemolytic agent, and at least including anion surfactant and pyrithione, the ratio between the content of anion surfactant and the content of pyrithione are 1.5~3:1.The hemolytic agent solves the not enough problem with excess of anion surfactant dosage, improves the accuracy of content of hemoglobin measure.
Description
Technical field
Reagent technique field is used the invention belongs to medical test, and in particular to a kind of blood cell analysis hemolytic agent.
Background technology
Hemolytic agent is one of the most frequently used reagent of blood cell analyzer, is also leucocyte and each point of group's counting, HGB contents
The key reagents of measure.After the blood being diluted adds hemolytic agent, erythrocytolysis discharges hemoglobin, the latter and hemolytic agent
Middle Related Component combines to form haemoglobin dervative, into 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 leucocyte (lymphocyte, monocyte, neutrophil leucocyte etc.) forms obvious difference after being acted on through hemolytic agent,
According to these differences, the leucocyte in blood sample can be divided into several monoids by instrument, and to total white blood cells and each monoid
Quantity counted.
At present, commonly used in domestic and international clinical examination and used without cyaniding hemolytic agent replacement KCN.Disclose in the prior art various
For the reagent of leukocyte differential count, such as CN2011100744709 be related to a kind of hemolytic agent for the differential counting of leucocyte five and
Purposes.The hemolytic agent is made up of hemolytic agent A with hemolytic agent B.Hemolytic agent A by polyoxyethylene-type nonionic surfactant, have
Machine acid, quaternary cationics cosolvent, stabilizer and osmotic pressure regulator composition；Hemolytic agent B is by basic mineral
Salt and osmotic pressure regulator are constituted.The hemolytic agent can by leucocyte be divided into lymphocyte, monocyte, eosinophil,
Neutrophil cell and basophilic granulocyte, obtain all kinds of quantity of leucocyte.
Anion surfactant can be acted on hemoglobin, form the coloured derivative of stabilization, and crest is 538nm,
Content of hemoglobin is directly proportional to absorbance at 538nm, so as to determine content of hemoglobin.But, such hemolytic agent is still
There is a problem of that measured value accuracy is low.
The content of the invention
The present invention proposes a kind of blood cell analysis hemolytic agent, and the hemolytic agent solves anion surfactant dosage not
Foot and excessive problem, improve the accuracy of content of hemoglobin measure.
The technical proposal of the invention is realized in this way：
A kind of blood cell analysis hemolytic agent, at least including anion surfactant and pyrithione, anion table
The ratio between the content of face activating agent and the content of pyrithione are 1.5~3:1.
Preferably, anion surfactant is lauryl sodium sulfate, pyrithione is sodium pyrithione.
Further, in some embodiments of the present invention, also contain buffer solution, the pH of buffer regulation hemolytic agent for 6~
8。
Further, the buffer solution is selected from malic acid, succinic acid, phthalic acid, citric acid, tartaric acid, oxalic acid, first
Acid, maleic acid, Gly and their own salt, and hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid.
Further, the buffer solution is disodium hydrogen phosphate and sodium oxalate.
Further, to eliminate the influence that anion surfactant brings bubble, some embodiments of the present invention also include
Defoamer, defoamer of the present invention is conventional defoamer in hemolytic agent, and its content reaches elimination bubble purpose on a small quantity, generally
0.5~2g/L.
Lauryl sodium sulfate, white or faint yellow powdery, is dissolved in water, insensitive to alkali and hard water；With decontamination, emulsification
With excellent foaming power；It is a kind of anion surfactant of poison micro- to human body.Its biological degradability>90%.Purposes：It is used as
Emulsifying agent, extinguishing chemical, foaming agent and textile auxiliary；Also serve as toothpaste and paste, powdery, the foaming agent of shampoo.
Sodium pyrithione, molecular formula C5H4NOSNa, is a kind of antimildew and antibacterial raw produce.Sterling is white or off-white color powder
The organic solvents such as end, soluble in water and ethanol.It is generically configured to 40% liquor, in faint yellow to yellowish-brown transparency liquid, easily
It is dissolved in water.Declined using holding effect in acid condition, stablized under alkaline or neutral conditions.
WBC is leucocyte (English name in the present invention：Leukocyte, white blood cell, referred to as：WBC), it is once called as
White blood cell.
In the present invention RBC be red blood cell also known as red blood cell or red blood cell, be most a kind of haemocytes in blood.It is red thin
Contain hemoglobin in born of the same parents, thus blood is taken on a red color.Hemoglobin can be combined with the oxygen in air, therefore red blood cell can pass through
The oxygen sucked in alveolar is shipped for tissue by hemoglobin, and the carbon dioxide that metabolism is produced in organizing is also by red blood cell
Transport to lung and be excreted.
HGB (hemoglobin in the present invention；haemoglobin；Hb；HGB) each haemoglobin molecule is by tetramolecular
Globin and four molecule hemes are constituted, and each ferroheme is made up of 4 pyrrole rings again, has an iron original in ring center
Son.Iron in hemoglobin can be combined (oxyhemoglobin), if iron is oxidized to three with oxygen in divalent state in invertibity
Valency state.
PLT (bloodplatelet) is blood platelet in the present invention, is one of visible component in mammalian, is
From the fritter kytoplasm with bioactivity that the ripe megacaryocyte kytoplasm crack releasing of marrow gets off.
Hemolytic agent is sold in market, adds anion surfactant particularly SDS (lauryl sodium sulfate), but SDS dosages
It is difficult to hold, because when HC is too high in blood sample, if SDS dosages are not enough, SDS is combined not with hemoglobin
Completely, content of hemoglobin is caused to determine inaccurate, if SDS dosage excess, meeting dialogue eucaryotic cell structure is damaged, and is changed
Bleach cell volume, so as to influence white blood cell count(WBC) and classification.Inventor is surprised to find pyrithione by lot of experiments
Can also be acted on hemoglobin, form the coloured derivative of stabilization, crest is 538nm, is inhaled at content of hemoglobin and 538nm
Shading value is directly proportional, so as to determine content of hemoglobin.Additionally, pyrithione dialogue eucaryotic cell structure and volume do not influence,
White blood count and differential is not interfered with.Appropriate pyrithione is added, the not enough problem with excess of SDS dosages is solved,
Ensure that the accuracy that content of hemoglobin is determined.
Specific embodiment
Embodiment 1
A kind of blood cell analysis hemolytic agent, with following components：
Lauryl sodium sulfate 10g/L, disodium hydrogen phosphate 1g/L, sodium oxalate 0.8g/L, sodium pyrithione 5g/L, defoamer
2g/L。
By each component precise as requested, with mixing, constant volume after water dissolves, then with 2.2 μm of membrane filtrations packing.
Embodiment 2
A kind of blood cell analysis hemolytic agent, with following components：
Lauryl sodium sulfate 12g/L, disodium hydrogen phosphate 1g/L, sodium oxalate 0.8g/L, pyrithione potassium 6g/L, defoamer
2g/L。
By each component precise as requested, with mixing, constant volume after water dissolves, then with 2.2 μm of membrane filtrations packing.
Embodiment 3
A kind of blood cell analysis hemolytic agent, with following components：
Lauryl sodium sulfate 12g/L, disodium hydrogen phosphate 1g/L, sodium oxalate 0.8g/L, pyrithione potassium 5g/L, defoamer
2g/L。
By each component precise as requested, with mixing, constant volume after water dissolves, then with 2.2 μm of membrane filtrations packing.
Test example
Hemolytic agent is sold by the hemolytic agent of embodiment 1 and market and compare research, determine haemolysis speed, hemoglobin
(HGB), the parameter such as leucocyte (WBC), red blood cell (RBC), hemoglobin (HGB), blood platelet (PLT).
1 materials and methods
1.1 instruments
1.1.1 auspicious BC-3000 cellanalyzers are stepped
1.2 reagents
1.1.1 auspicious BC-3000 cellanalyzers genuine matched reagent is stepped：Dilution, genuine hemolytic agent, cleaning fluid.
1.2.2 the hemolytic agent of embodiment 1 (hemolytic agent of abbreviation example 1).
1.2.3 the clinical patient venous blood of sample experiment same day addition EDTA-K2 anti-freezings, has detected after blood sampling in 4 hours
Finish.
1.2.4 Quality Control blood：Sichuan mikey (lot number：1509176).
1.3 experimental techniques
1.3.1 haemolysis speed observer
Laboratory temperature sets 20 DEG C, takes 10ml colorimetric cylinders 2, adds dilution 5ml, the μ l of same anticoagulation 20, then
Genuine hemolytic agent and each 1ml of invention hemolytic agent are separately added into, and are shaken up, erythrocytolysis situation is observed immediately, record red blood cell
It is completely dissolved the time.
20 normal specimens and 20 monstrosities are extracted, above-mentioned experiment is carried out.
1.3.2 clinical detection
Laboratory temperature sets 20 DEG C before experiment, and Quality Control blood is determined on BC-3000 cellanalyzers, detects projects
Whether in reference range, it is ensured that instrument normal work.
20 normal specimens and 20 monstrosities are extracted, it is thin in blood using genuine hemolytic agent and hemolytic agent of the present invention respectively
Detected on born of the same parents' analyzer, every testing result such as record HGB, WBC, RBC, PLT.
By cervical arthroplasty mode carry out detect above-mentioned 20 normal specimens and 20 monstrosities, record HGB, WBC,
The items testing result such as RBC, PLT.
In cervical arthroplasty mode as the standard value of each detection project.
2 results and analysis
2.1 haemolysis speed
Genuine hemolytic agent and the hemolytic agent haemolysis speed of embodiment 1 are shown in Tables 1 and 2.
The normal specimen genuine hemolytic agent of table 1 and the hemolytic agent haemolysis speed of example 1 are compareed
The monstrosity genuine hemolytic agent of table 2 and the hemolytic agent haemolysis speed of example 1 are compareed
As it can be seen from table 1 the genuine hemolytic agent of normal specimen and the hemolytic agent haemolysis speed of embodiment 1 are essentially identical,
It is completely dissolved red blood cell in 8-10s.
From table 2 it can be seen that the genuine hemolytic agent haemolysis speed of monstrosity is in 7-12s, the hemolytic agent haemolysis of embodiment 1
Speed is in 8-10s, and the hemolytic agent of example 1 is more more stable than genuine hemolytic agent for the monstrosity hemolysis time.
2.2 clinical detection results and analysis
Respectively with genuine hemolytic agent, 20 normal specimens of the hemolytic agent of example 1 and cervical arthroplasty mode clinical assays and 20
Monstrosity, major parameter statistics is shown in Table 3 and table 4.
3 normal specimen of table, two kinds of reagents and cervical arthroplasty clinical assays results contrast
4 monstrosity of table, two kinds of reagents and cervical arthroplasty clinical assays results contrast
From table 3 it is observed that two kinds of reagents of normal specimen HGB testing results are basically identical with cervical arthroplasty, without significantly
Sex differernce.
As can be seen from Table 4, monstrosity HGB testing results, genuine hemolytic agent determines HGB to be had significantly with cervical arthroplasty
Sex differernce, the hemolytic agent of embodiment 1 determines HGB, and there was no significant difference with cervical arthroplasty, and invention hemolytic agent clinical detection result more connects
Nearly actual value.
Presently preferred embodiments of the present invention is the foregoing is only, is not intended to limit the invention, it is all in essence of the invention
Within god and principle, any modification, equivalent substitution and improvements made etc. should be included within the scope of the present invention.
Claims (6)
1. a kind of blood cell analysis hemolytic agent, it is characterised in that at least including anion surfactant and pyrithione,
The ratio between the content of anion surfactant and the content of pyrithione are 1.5~3:1.
2. blood cell analysis hemolytic agent according to claim 1, it is characterised in that anion surfactant is 12
Sodium alkyl sulfate, pyrithione is sodium pyrithione.
3. blood cell analysis hemolytic agent according to claim 1 and 2, it is characterised in that also contain buffer solution, the buffering
The pH of agent regulation hemolytic agent is 6~8.
4. blood cell analysis hemolytic agent according to claim 3, it is characterised in that the buffer solution be selected from malic acid,
Succinic acid, phthalic acid, citric acid, tartaric acid, oxalic acid, formic acid, maleic acid, Gly and their own salt, and salt
Acid, sulfuric acid, phosphoric acid, nitric acid.
5. blood cell analysis hemolytic agent according to claim 4, it is characterised in that the buffer solution is disodium hydrogen phosphate
With sodium oxalate.
6. blood cell analysis hemolytic agent according to claim 1 and 2, it is characterised in that also including defoamer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710174670.9A CN106932350A (en) | 2017-03-22 | 2017-03-22 | Blood cell analysis hemolytic agent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710174670.9A CN106932350A (en) | 2017-03-22 | 2017-03-22 | Blood cell analysis hemolytic agent |
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| Publication Number | Publication Date |
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| CN106932350A true CN106932350A (en) | 2017-07-07 |
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ID=59432014
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710174670.9A Pending CN106932350A (en) | 2017-03-22 | 2017-03-22 | Blood cell analysis hemolytic agent |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108872225A (en) * | 2018-07-17 | 2018-11-23 | 浙江亚培生物技术有限公司 | A kind of detection reagent and detection method detecting animal blood cell |
| CN111781047A (en) * | 2020-07-03 | 2020-10-16 | 四川恒健生物科技有限公司 | Hemolysin for blood analysis and preparation method and reagent thereof |
Citations (5)
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|---|---|---|---|---|
| CN101464453A (en) * | 2007-12-18 | 2009-06-24 | 深圳迈瑞生物医疗电子股份有限公司 | Hemolytic agent |
| CN101819199A (en) * | 2010-05-08 | 2010-09-01 | 桂林市朗道诊断用品有限公司 | Reagent for hemocyte analyzers |
| CN102662067A (en) * | 2012-05-20 | 2012-09-12 | 烟台卓越生物技术有限责任公司 | Cyanide-free hemolysin for determining hemoglobin concentration and carrying out white blood cell count by groups |
| CN103323582A (en) * | 2013-06-18 | 2013-09-25 | 南京普朗医疗设备有限公司 | Leukocyte classification hemolytic agent and kit thereof |
| CN103698501A (en) * | 2013-12-23 | 2014-04-02 | 深圳市开立科技有限公司 | Cyanide-free hemolytic agent |
-
2017
- 2017-03-22 CN CN201710174670.9A patent/CN106932350A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101464453A (en) * | 2007-12-18 | 2009-06-24 | 深圳迈瑞生物医疗电子股份有限公司 | Hemolytic agent |
| CN101819199A (en) * | 2010-05-08 | 2010-09-01 | 桂林市朗道诊断用品有限公司 | Reagent for hemocyte analyzers |
| CN102662067A (en) * | 2012-05-20 | 2012-09-12 | 烟台卓越生物技术有限责任公司 | Cyanide-free hemolysin for determining hemoglobin concentration and carrying out white blood cell count by groups |
| CN103323582A (en) * | 2013-06-18 | 2013-09-25 | 南京普朗医疗设备有限公司 | Leukocyte classification hemolytic agent and kit thereof |
| CN103698501A (en) * | 2013-12-23 | 2014-04-02 | 深圳市开立科技有限公司 | Cyanide-free hemolytic agent |
Non-Patent Citations (1)
| Title |
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| 雷万军 等: "《医学检验专业实验指导》", 30 September 2012, 郑州大学出版社 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108872225A (en) * | 2018-07-17 | 2018-11-23 | 浙江亚培生物技术有限公司 | A kind of detection reagent and detection method detecting animal blood cell |
| CN111781047A (en) * | 2020-07-03 | 2020-10-16 | 四川恒健生物科技有限公司 | Hemolysin for blood analysis and preparation method and reagent thereof |
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