CN108254579A - Prepare a kind of high-density lipoprotein cholesterol detection kit of strong antijamming capability - Google Patents
Prepare a kind of high-density lipoprotein cholesterol detection kit of strong antijamming capability Download PDFInfo
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- CN108254579A CN108254579A CN201810093254.0A CN201810093254A CN108254579A CN 108254579 A CN108254579 A CN 108254579A CN 201810093254 A CN201810093254 A CN 201810093254A CN 108254579 A CN108254579 A CN 108254579A
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- reagent
- kit
- antijamming capability
- density lipoprotein
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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
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/92—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving lipids, e.g. cholesterol, lipoproteins, or their receptors
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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
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/5306—Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/32—Cardiovascular disorders
- G01N2800/323—Arteriosclerosis, Stenosis
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/32—Cardiovascular disorders
- G01N2800/324—Coronary artery diseases, e.g. angina pectoris, myocardial infarction
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Abstract
The present invention prepares a kind of high-density lipoprotein cholesterol detection kit of strong antijamming capability, its significance lies in that providing a kind of stability and antijamming capability enhancing, storing and convenient transportation, accuracy and the high high-density lipoprotein cholesterol detection kit of precision.Kit of the present invention is a kind of liquid-type double reagent vitro detection kit, reagent is mainly made of reagent R1 and reagent R2, the present invention is on the basis of conventional formulation, by introducing the substances such as chloroform, protamine sulfate, myristyl betaine, mannitol and glycerine in reagent, the change of matter is made that reagent, makes it have stronger stability and antijamming capability.
Description
Technical field
The present invention relates to clinical in vitro diagnosis in vitro reagent technique field, the high density fat of more particularly to a kind of strong antijamming capability
Protein cholesterol detection kit.
Background technology
Presence and types of transportation of the plasma lipoprotein as cholesterol in blood plasma, i.e., nonpolar lipid includes courage in blood plasma
Sterol and hydrophilic apolipoprotein combine so that lipoprotein has stronger water solubility that can be transported in blood.High density lipoprotein level
In vain, low-density lipoprotein, very low density lipoprotein and chylomicrons together constitute plasma lipoprotein, wherein low-density lipoprotein
Be mainly responsible for cholesterol transport to peripheral tissues, and high-density lipoprotein be remove arterial wall on cholesterol to liver transport
It is defeated.The large-scale epidemiological surveys of Frammingham in 1977 show that plasma hdl cholesterol decline is scarce
Courageous and upright cardiopathic Major Risk Factors, henceforth, the measure of HDL-C is increasingly subject to most attention clinically.HDL is
As the prevention factor of the heart disease caused by artery sclerosis, therefore the measure of HDL-C has in terms of arteriosclerosis morbidity
Important predicting function, this clinically has great importance.Stream medical research shows low-density lipoprotein cholesterol level
It is positively correlated with the incidence of artery sclerosis class disease coronary heart disease, high-density lipoprotein cholesterol and artery sclerosis class disease coronary disease
The incidence of disease is negatively correlated, i.e., with anti arteriosclerosis activity.High-density lipoprotein cholesterol HDL-C, low-density lipoprotein
The risk factor that cholesterol LDL-C is often used as the cardiovascular and cerebrovascular disease of most worthy in clinical labororatory's lipid determination refers to
Mark.
Presently commercially available similar product, usually enzyme process detection kit, but such kit generally existing stability
Can be poor, antijamming capability is weak the problems such as.Based on this, kit of the present invention is intended to improve what is in conventional reagents box detection process occurred
Deficiency, and traditional enzyme process kit is modified.
Invention content
The present invention prepares a kind of high-density lipoprotein sterol detection kit of strong antijamming capability, kit master of the present invention
If by introducing several substances such as chloroform, protamine sulfate, myristyl betaine, mannitol and glycerine, first, increasing
The strong antijamming capability of reagent, second is that there is new raising to the stability of reagent.It is red that chloroform can effectively remove courage
Element is to interference caused by testing result；Protamine sulfate can be effectively in macromolecular substances（Such as heparin）, for macromolecular
Result caused by substance is disorderly, and protamine sulfate is fully able to solve the problems, such as such result disorder；Myristyl betaine
It introduces, on the one hand increases reagent reaction rate, effective guarantor on the other hand is both provided to the long-term storage and transport of reagent
Shield acts on；The introducing of glycerine and mannitol increases the stability of reagent.
Description of the drawings
Fig. 1 is kit of the present invention and contrast agents box Detection of Stability result
Fig. 2 is that kit of the present invention stores collection of illustrative plates with contrast agents box under the conditions of 4 DEG C
Fig. 3 is that kit of the present invention stores collection of illustrative plates with contrast agents box under the conditions of 37 DEG C
Fig. 4 is the antijamming capability testing result of kit of the present invention and contrast agents box
Fig. 5 present invention and contrast agents box repetitive test result
Fig. 6 is kit of the present invention and contrast agents box clinical detection result
Fig. 7 is kit of the present invention and contrast agents box correlation collection of illustrative plates.
Specific embodiment
Example 1
The reagent R1 is formed：
Polyanion cellulose 0.5mmol/L
Cholesterol oxidase 0.8KU/L
Peroxidase 1.5KU/L
TTAB 0.25mmol/L
PIPES buffer solution 50m mol/L
Myristyl betaine 0.5g/L
Chloroform 5ml/L
Protamine sulfate 5g/L
Reagent R2 is formed：
4- antipyrine 25m mol/L
Cholesteryl esterase 3KU/L
Tween-20 2ml/L
Mannitol 10g/L
Glycerine 5ml/L
GOODS buffer solution 75m mol/L.
Example 2
Reagent R1 is formed：
Polyanion cellulose 0.5mmol/L
Cholesterol oxidase 0.8KU/L
Peroxidase 1.5KU/L
TTAB 0.25mmol/L
PIPES buffer solution 50m mol/L
Myristyl betaine 1.5g/L
Chloroform 2.5ml/L
Protamine sulfate 5g/L
Reagent R2 is formed：
4- antipyrine 25m mol/L
Cholesteryl esterase 3KU/L
Tween-20 5ml/L
Mannitol 10g/L
Glycerine 2ml/L
GOODS buffer solution 75m mol/L.
Example 3
Reagent R1 is formed：
Polyanion cellulose 0.5mmol/L
Cholesterol oxidase 0.8KU/L
Peroxidase 1.5KU/L
TTAB 0.25mmol/L
PIPES buffer solution 50m mol/L
Myristyl betaine 3.5g/L
Chloroform 3.5ml/L
Protamine sulfate 7.5g/L
Reagent R2 is formed：
4- antipyrine 25m mol/L
Cholesteryl esterase 3KU/L
Tween-20 2-5ml/L
Mannitol 10g/L
Glycerine 5ml/L
GOODS buffer solution 75m mol/L.
Example 4
Reagent R1 is formed：
Polyanion cellulose 0.5mmol/L
Cholesterol oxidase 0.8KU/L
Peroxidase 1.5KU/L
TTAB 0.25mmol/L
PIPES buffer solution 50m mol/L
Myristyl betaine 5g/L
Chloroform 5ml/L
Protamine sulfate 7.0g/L
Reagent R2 is formed：
4- antipyrine 25m mol/L
Cholesteryl esterase 3KU/L
Tween-20 3.0ml/L
Mannitol 5g/L
Glycerine 4.5ml/L
GOODS buffer solution 75m mol/L.
Kit performance detection experiment of the present invention
Stability test detects
With kit of the present invention（The formula of embodiment 1 is prepared）Two kits to circulate currently on the market are compared, school
Quasi- product and quality-control product are institute's band in contrast agents, the use of instrument are automatic clinical chemistry analyzer BS-800, kit ginseng of the present invention
Number setting is as follows, and dominant wavelength 546nm, read point is set as 14-16,32-33, calibrates as 2 terminals, and as a result unit is mmol/
L, sample size：Reagent R1：Reagent R2=3ul：225ul：75ul, contrast agents box are carried out in strict accordance with kit operational manual,
Detecting instrument is BS-800, rear to carry out result detection, and testing result refers to Fig. 1.
Pass through Fig. 1, Fig. 2 and Fig. 3, it has been found that by the stability tests of 120 days with monitoring, it has been found that in 4 DEG C of items
Under part, after kit of the present invention lasts 120 days, testing result and quality-control product mark target value deviation less than 5%, but contrast agents
After box A and contrast agents box B lasts 120 days and quality-control product mark target value deviation has exceeded 25%；Present invention examination under the conditions of 37 DEG C
Testing result and quality-control product mark target value deviation less than 5% after agent box lasts 120 days, relative to contrast agents A and contrast agents B
For, 10% or so, contrast agents box B was examined at the 4th day for testing result and quality-control product deviation after contrast agents A lasts 30 days
It surveys result and has deviated from quality-control product 10%.By data above we can see that kit of the present invention carrying in terms of stability
It is high.
Anti-interference test
We buy similar kit in current circulation market as contrast agents, and quality-control product is as standard substance.Purchase
Bilirubin, heparin, hemoglobin, ascorbic acid and artificial serum, are separately added into bilirubin in artificial serum, and heparin is blood red
Albumen, four kinds of substances of ascorbic acid are bilirubin concentration 300umol/L, heparin 40mg/L, hemoglobin 20mg/L, anti-bad
Hematic acid 30mg/L.It is tested with BS-800 automatic clinical chemistry analyzers, kit parameter setting of the present invention is as follows, dominant wavelength
For 546nm, read point is set as 14-16,32-33, calibrates as 2 terminals, and as a result unit is mmol/L, sample size：Reagent R1：
Reagent R2=3ul：225ul：75ul, contrast agents box are carried out in strict accordance with kit operational manual, are formulated using embodiment 2,
Test result refers to Fig. 4.
By Fig. 4, we can be found that kit of the present invention for bilirubin, heparin, hemoglobin, four kinds of ascorbic acid
The antijamming capability of interfering substance is better than contrast agents box A and contrast agents box B.
Repetitive test
With real kit of the invention（The formula of embodiment 3 is prepared）Two kits to circulate currently on the market are compared,
Calibration object and quality-control product are institute's band in contrast agents, the use of instrument are automatic clinical chemistry analyzer BS-800, to contrast agents box
Charge product（It accuses flat comprising two kinds of high level and low value quality-control product）Carry out N=20 time detection, the coefficient of variation of analytical reagent.Detection
The results detailed in Fig. 5.
By Fig. 5, we are evident that, three group reagents are compared, and kit of the present invention either high level quality-control product is still
Low value quality-control product, testing result show that the coefficient of variation is below contrast agents box.
Comparability test
Kit of the present invention tests random N=40 sample with contrast agents box, and test result exists in table form
Following table is embodied, the numerical value recorded by the table 4, and then is calculated, the kit of kit of the present invention and market circulation
Correlation.Kit of the present invention is detected using the formula of embodiment 4.Testing result refers to Fig. 6.
Pass through Fig. 6 and Fig. 7, it has been found that kit of the present invention and contrast agents box carry out clinical random sample number N=40
New technical conditions have been established in detection, correlation coefficient r >=0.998 of the two so as to put goods on the market for kit of the present invention.
Claims (3)
1. a kind of high-density lipoprotein cholesterol detection kit of strong antijamming capability, it is characterised in that：The kit is
A kind of liquid-type double reagent vitro detection kit being made of reagent R1 and reagent R2, the reagent R1 are included：Chelating agent, courage
Sterol oxidizing ferment, peroxidase, surfactant, buffer solution, stabilizer, chloroform and protamine sulfate；Reagent R2 is included：
4- antipyrines, cholesteryl esterase, surfactant, stabilizer and buffer solution.
2. kit according to claim 1, it is characterised in that：Chloroform and protamine sulfate tool in the reagent R1
There is very strong antijamming capability.
3. kit according to claim 1, it is characterised in that：Stabilizer in the reagent R1 is myristyl beet
Alkali；Stabilizer in the reagent R2 includes mannitol and glycerine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810093254.0A CN108254579B (en) | 2018-01-31 | 2018-01-31 | Preparation of high-density lipoprotein cholesterol detection kit with strong anti-interference capability |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810093254.0A CN108254579B (en) | 2018-01-31 | 2018-01-31 | Preparation of high-density lipoprotein cholesterol detection kit with strong anti-interference capability |
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| Publication Number | Publication Date |
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| CN108254579A true CN108254579A (en) | 2018-07-06 |
| CN108254579B CN108254579B (en) | 2020-10-30 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112695071A (en) * | 2020-12-16 | 2021-04-23 | 浙江伊利康生物技术有限公司 | High-density lipoprotein 3 determination reagent, method and kit |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002063302A1 (en) * | 2001-02-07 | 2002-08-15 | Immunomatrix Inc. | A method for reducing heparin interference in diagnostic tests for cardiac troponin |
| CN1379235A (en) * | 2002-05-10 | 2002-11-13 | 肖洪武 | Process and reagent for measuring high-density lipoprotein and cholesterol |
| CN101078729A (en) * | 2006-05-10 | 2007-11-28 | 富士胶片株式会社 | Method for measuring high density lipoprotein cholesterol |
| CN104076158A (en) * | 2014-07-10 | 2014-10-01 | 深圳市新产业生物医学工程股份有限公司 | Kit for measuring high-density lipoprotein cholesterol |
| CN106244671A (en) * | 2016-08-30 | 2016-12-21 | 山东博科生物产业有限公司 | A kind of stable aspartate aminotransferase checkout and diagnosis detection kit |
| CN106908612A (en) * | 2015-12-22 | 2017-06-30 | 山东博科生物产业有限公司 | A kind of albumin detection reagent box of antiheparin stabilization |
-
2018
- 2018-01-31 CN CN201810093254.0A patent/CN108254579B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002063302A1 (en) * | 2001-02-07 | 2002-08-15 | Immunomatrix Inc. | A method for reducing heparin interference in diagnostic tests for cardiac troponin |
| CN1379235A (en) * | 2002-05-10 | 2002-11-13 | 肖洪武 | Process and reagent for measuring high-density lipoprotein and cholesterol |
| CN101078729A (en) * | 2006-05-10 | 2007-11-28 | 富士胶片株式会社 | Method for measuring high density lipoprotein cholesterol |
| CN104076158A (en) * | 2014-07-10 | 2014-10-01 | 深圳市新产业生物医学工程股份有限公司 | Kit for measuring high-density lipoprotein cholesterol |
| CN106908612A (en) * | 2015-12-22 | 2017-06-30 | 山东博科生物产业有限公司 | A kind of albumin detection reagent box of antiheparin stabilization |
| CN106244671A (en) * | 2016-08-30 | 2016-12-21 | 山东博科生物产业有限公司 | A kind of stable aspartate aminotransferase checkout and diagnosis detection kit |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112695071A (en) * | 2020-12-16 | 2021-04-23 | 浙江伊利康生物技术有限公司 | High-density lipoprotein 3 determination reagent, method and kit |
| CN112695071B (en) * | 2020-12-16 | 2022-12-30 | 浙江伊利康生物技术有限公司 | High-density lipoprotein 3 determination reagent, method and kit |
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| CN108254579B (en) | 2020-10-30 |
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Effective date of registration: 20200927 Address after: 250100 Mingshui Economic Development Zone, Zhangqiu District, Jinan City, Shandong Province Applicant after: BIOBASE BIODUSTRY (SHANDONG) Co.,Ltd. Address before: 250200 2 unit 201, Boko Industrial Park, north of Ji Wang Road, Zhangqiu City, Ji'nan, Shandong. Applicant before: JINAN JINGZHENG MEDICAL EQUIPMENT Co.,Ltd. |
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