CN108841918A - A kind of sdLDL-C kit - Google Patents
A kind of sdLDL-C kit Download PDFInfo
- Publication number
- CN108841918A CN108841918A CN201810034866.2A CN201810034866A CN108841918A CN 108841918 A CN108841918 A CN 108841918A CN 201810034866 A CN201810034866 A CN 201810034866A CN 108841918 A CN108841918 A CN 108841918A
- Authority
- CN
- China
- Prior art keywords
- reagent
- buffer
- sdldl
- content
- cholesterol
- 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.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/60—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving cholesterol
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/26—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/26—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase
- C12Q1/28—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase involving peroxidase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/26—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase
- C12Q1/30—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase involving catalase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
- C12Q1/40—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving amylase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
- C12Q1/44—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving esterase
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/902—Oxidoreductases (1.)
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/902—Oxidoreductases (1.)
- G01N2333/908—Oxidoreductases (1.) acting on hydrogen peroxide as acceptor (1.11)
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/914—Hydrolases (3)
- G01N2333/916—Hydrolases (3) acting on ester bonds (3.1), e.g. phosphatases (3.1.3), phospholipases C or phospholipases D (3.1.4)
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/914—Hydrolases (3)
- G01N2333/916—Hydrolases (3) acting on ester bonds (3.1), e.g. phosphatases (3.1.3), phospholipases C or phospholipases D (3.1.4)
- G01N2333/918—Carboxylic ester hydrolases (3.1.1)
- G01N2333/92—Triglyceride splitting, e.g. by means of lipase
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a kind of sdLDL-C kits, it is made of two independent reagent R1 and reagent R2, R1 contains buffer, lipoprotein lipase, nonionic surfactant, seralbumin, heparin sodium, magnesium ion, cholesterol esterase, cholesterol oxidase, catalase or peroxidase, Trinder ' s reactant A, stabilizer, preservative;R2 contains buffer, nonionic surfactant, Trinder ' s reactant B, Sodium azide, preservative.The cholesterol eliminated other than SD-LDLC is first mixed with sample with reagent R1, is reacted after reagent R2 addition for SD-LDLC, and Trinder ' s reactant B colour generation measures absorbance, calculates its content.Stabilization of kit of the present invention is good, and accuracy is high, easy to operate quick, is suitable for automatic clinical chemistry analyzer and detects.
Description
Technical field
The present invention relates to a kind of sdLDL-C kits.
Background technique
《Chinese cardiovascular disease report 2016》Point out, about 2.9 hundred million people of China's cardiovascular patient number, occupy people's death rate it
40% or more, and Blood Cholesterol amount is too high, especially bad cholesterol --- low density lipoprotein cholesterol (Low
Density Lipoprotein Cholesterol, abbreviation LDLC) it is excessively high be the one of the main reasons for causing cardiovascular disease.
In blood in addition to a small amount of free cholesterol, granule is formed in conjunction with most cholesterol protein corresponding to its
Product is different, density difference lipoprotein cholesterol, in clinical examination common ultracentrifugation be classified as chylomicron (Chylomicron,
Abbreviation CM, 80~500nm of granular size, density < 0.95g/L), very low density lipoprotein (Very Low Density
Lipoprotein, abbreviation VLDL, 30~80nm of granular size, 0.95~1.006g/L of density) cholesterol, low-density lipoprotein
(Low Density Lipoprotein, abbreviation LDL, 19~30nm of granular size, 1.019~1.063g/L of density) cholesterol,
And high-density lipoprotein (High Density Lipoprotein, abbreviation HDL, 7~10nm of granular size, density 1.063~
1.21g/L) cholesterol;Wherein HDL can decompose the cholesterol transport in arterial wall to liver metabolism to reduce cholesterol in blood
The deposition of tube wall plays the role of antiatherosclerosis and protection blood vessel, and in contrast, LDL then consolidates the gallbladder in blood
Alcohol, which is transported to deposition in cells of vascular wall, leads to atheroma, is the high-risk factor of cardiovascular disease.
The compositions such as cholesterol, phosphatide of the LDL of each molecule containing an apolipoprotein B (APOB) and its connection, will purify
The further ultracentrifugation of LDL can be separated into two parts, particle is larger, density relatively light (1.019~1.044g/L) title
Gently (big) low-density lipoprotein (Light or Large LDL, abbreviation L-LDL) and small volume, density it is larger (1.044~
Title small dense low-density lipoprotein (Small Dense Low Density Lipoprotein, abbreviation SD- 1.063g/L)
LDL), SD-LDL due to particle it is small, easily infiltrate into arterial blood tube wall, lower ldl receptor (LDL Receptor) is affine
Power, is not easy to be metabolized, and has longer half-life period in blood, and oxidation resistance ratio L-LDL is low, cardiovascular disease it is pathogenic
Than 3.5 times of L-LDL high or more;Because SD-LDL is VLDL (abbreviation VLDL1) the metabolism generation by being rich in triglycerides, and VLDL1
Stimulation in liver synthesis by insulin resistance (Insulin resistance), especially its pancreas of diabetes B patient
The increase of island element resistance results in the increase of SD-LDL, so SD-LDL other than the good index of cardiovascular disease, is also 2 types sugar
Urinate the important diagnostic factor of disease.
In addition, the incidence discovery of Lamarche et al. tracking 2013 normal persons (59 ± 7 years old) ischemic heart disease (IHD)
Behind 5 years of SD-LDL high, the disease incidence of IHD is higher by 3.6~5.1 times, therefore SD-LDL will become the significant terms of clinical diagnosis
Mesh.
SD-LDL measures common gradient gel electrophoresis (Gradient gel electrophoresis), proton magnetic resonance (PMR)
(Proton NMR spectroscopy), gradient ultra centrifugation (Density gradient ultracentrifugation),
High pressure liquid phase (HPLC) etc. measures, and equipment is expensive, complicated for operation, is unsuitable for routine inspection measurement and uses, though Hirano et al.
Invention can be measured with turbidimetry reagent with automatic clinical chemistry analyzer, but sample must be handled first, by the rouge egg other than SD-LDL
After white centrifugation, then by supernatant with turbidimetry for Determination, troublesome in poeration, specificity and accuracy be not high.
Summary of the invention
Place that purpose of the invention is to overcome the shortcomings in the prior art, it is simple to provide a kind of component, proportion rationally,
Stability is good, and accuracy is high, easy to operate quick, the small dense low-density lipoprotein suitable for automatic clinical chemistry analyzer detection
Cholesterol Kit.
A kind of sdLDL-C kit, it is characterised in that including independent reagent R1 and reagent
R2,
It wherein include buffer, lipoprotein lipase, cholesterol esterase, cholesterol oxidase, heparin sodium, chlorine in reagent R1
Change magnesium, seralbumin, nonionic surfactant, catalase or peroxidase, preservative;
The reagent R2 includes buffer, nonionic surfactant, Trinder ' s reactant B, preservative.
A kind of sdLDL-C kit as described above, it is characterised in that further include in reagent R1
There is Trinder ' s reactant A.
A kind of sdLDL-C kit as described above, it is characterised in that further include having stabilization
Agent.
A kind of sdLDL-C kit as described above, it is characterised in that buffer in reagent R1
Concentration be 20~200mM, pH=6.0~7.5;The content of lipoprotein lipase is 0.1~2u/ml, cholesterol esterase contains
Amount is 0.5~2.0u/ml, the content of cholesterol oxidase is 0.3~1.5u/ml, the content of heparin sodium is 0.1~0.5g/L,
The concentration of magnesium chloride is 2~20mM, sero-abluminous content is 1~20g/L, the content of nonionic surfactant is 0.1
~2.0ml/L, catalase content be 50~200u/ml (or when using peroxidase to eliminate the H2O2 in R1, then
Peroxidase content is 0.3~3ku/L), the content of preservative be 0.005-0.105%.
A kind of sdLDL-C kit as described above, it is characterised in that the buffer is
Good ' s buffer, BES buffer, Bis-Tris buffer, Pipes buffer, Mops buffer, in HEPES buffer solution
It is a kind of.
A kind of sdLDL-C kit as described above, it is characterised in that the non-ionic surface
Activating agent is one or both of styrene polyoxyethylene ether, benzyl phenyl polyoxyethylene ether.
A kind of sdLDL-C kit as described above, it is characterised in that delay in the reagent R2
The concentration of fliud flushing is 20~200mM, pH=6.0~7.5;The mass concentration of the nonionic surfactant is 0.5~5%;
The concentration of Trinder ' s reactant B is 2-5mM, the content of preservative is 0.005-0.105%.
A kind of sdLDL-C kit as described above, it is characterised in that delay described in reagent R2
Fliud flushing is Good ' s buffer, BES buffer, Bis-Tris buffer, Pipes buffer, Mops buffer, HEPES buffering
One of liquid.
A kind of sdLDL-C kit as described above, it is characterised in that non-described in reagent R2
Ionic surface active agent be Pluronic F68 or alkyl phenol polyoxyethylene ether,
A kind of sdLDL-C kit as described above, it is characterised in that further include in reagent R2
Weight percent is the Sodium azide of 0.1-0.2%.
This kit is made of reagent 1 and reagent 2, and it is solid first to mix the gallbladder eliminated other than SD-LDLC with sample with R1 reagent
Alcohol reacts after reagent R2 addition for SD-LDLC, and Trinder ' s reactant B colour generation measures absorbance, calculates its content.
Use Good ' s buffer in reagent R1 of the present invention, 20~200mM of concentration, other than PH6.0~7.5, mainly with
Special component and its proper content eliminates the cholesterol other than SD-LDLC in advance.
Component in reagent R1 of the present invention includes 0.1~2u/ml of lipoprotein lipase, 0.1~0.5g/L of heparin sodium, chlorine
Change 2~20mM of magnesium, 1~20g/L of seralbumin, styrene 0.1~2.0ml/L of polyoxyethylene ether, benzyl phenyl polyoxyethylene
0.3~2.5ml/L of ether mixture or styrene polyoxyethylene and benzyl phenyl polyoxyethylene 0.1~2.5ml/L of ether mixture,
The HLB value of its mixture is controlled 12~14.It can be by the lipoprotein gallbladder other than SD-LDLC in the presence of described group point common
Sterol dissociation can be decomposed by cholesterol esterase and cholesterol oxidase and be eliminated.
H2O2 produced by cholesterol other than SD-LDLC is decomposed to eliminate with catalase in reagent R1 of the present invention, or
With peroxidase and Trinder ' s reactant A elimination H2O2.
Trinder ' s reactant A is one of HDAOS, DAOS, TODB, TOOS in reagent R1 of the present invention, to eliminate
The H2O2 of generation.
The nonionics such as Pluronic F68 or alkyl phenol polyoxyethylene ether are added in R2 reagent of the present invention
Surfactant, such as Pluronic F-88, Pluronic F-68, Pluronic 17R-4, Emulgen709, for dissolution
SD-LDLC makes it form H2O2 with cholesterol esterase and cholesterol oxidation enzyme reaction, then determines after reacting colour generation according to Trinder ' s
Measure it.
H2O2 is such as eliminated with catalase in reagent R1 in the present invention, then the Sodium azide containing 0.1-0.2% in R2 reagent.
Preservative described in reagent R1 and reagent R2 is methyl benzoic acid, ethyl benzoate, propylbenzene first in the present invention
The mixture of the one or two or more kinds such as acid, chloramphenicol, radioactive ray ketone, Kathon CG.
Heretofore described stabilizer is glycine or alanine or glutamic acid.
Trinder ' s reactant B is 4- amino antipyrine, abbreviation 4-AA in the present invention.
In the present invention in reagent R1 buffer (Lipoprotein Lipase) containing lipoprotein lipase to decompose blood
CM, VLDL of content of triglyceride height in sample, become fragment.
Contain appropriate seralbumin, heparin sodium and magnesium ion in the present invention in R1 reagent, protecting LDL-C to exempt from by CE and
CO is decomposed.
Styrene polyoxyethylene ether, benzyl phenyl polyoxyethylene ether in reagent R1 in the present invention, or both proper proportion it
Mixture can make the dissolution of the L-LDL in LDL that can be decomposed by CE and CO.
Cholesterol esterase is added in the present invention and cholesterol oxidase is used to decompose CM fragment, VLDL fragment, HDL and L-
LDL upper cholesterol generates H2O2;
H2O2 of the catalase to eliminate generation after CE and CO reacts is added in the present invention, or with peroxidase
H2O2 is eliminated with Trinder ' s reactant A;
Trinder ' s reactant A such as TODB, HDAOS, TOOS of the present invention are anti-in Catalyzed Synthesis By Peroxidase with hydrogen peroxide
Leuco-compounds should be formed to exclude hydrogen peroxide;
The nonionic surfactant that R2 reagent includes in the present invention can make it dissolve out SD-LDLC and react with CE and CO.
In conclusion beneficial effects of the present invention:
Kit of the invention is made of two independent reagent R1 and reagent R2, and R1 contains buffer, lipoprotein lipase
(Lipoprotein Lipase), nonionic surfactant, seralbumin, heparin sodium, magnesium ion, cholesterol esterase, gallbladder
Sterol oxidizing ferment, catalase or peroxidase, Trinder ' s reactant A, stabilizer, preservative;R2 containing buffer,
Nonionic surfactant, Trinder ' s reactant B, Sodium azide, preservative.After reagent R1 is mixed with detection sample, examination
Contained lipoprotein lipase will be rich in chylomicron (Chylomicron, the abbreviation of triglycerides in agent R1 in blood plasma (clear) sample
CM it) decomposes and fragmentates, in the presence of heparin sodium, magnesium ion and people or ox or other animal albumin and high-density lipoprotein gallbladder is solid
Alcohol (HDLC) first decomposes elimination, the low density lipoprotein cholesterol (LDL- of remainder by cholesterol esterase, cholesterol oxidase simultaneously
C) contain styrene polyoxyethylene ether, benzyl phenyl polyoxyethylene ether of debita spissitudo or both at the same time according to proper proportion mixture
In the presence of, further by bulky grain (or light) low-density lipoprotein (Large LDL or Light LDL, abbreviation in LDL
L-LDL) cholesterol (L-LDL-C) appropriate cholesterol esterase and cholesterol oxidase in the presence of decomposed, protect simultaneously
Small dense low-density lipoprotein (Small Dense Low Density Lipoprotein, abbreviation SD-LDL) has been stayed to exempt to be divided
Solution, the present invention are to decompose the cholesterol other than SD-LDLC contained in sample in advance using particular components contained in R1 to disappear
It removes, and remains SD-LDLC, after R2 addition, the SD-LDLC retained is in the presence of another nonionic surfactant
It is decomposed by cholesterol esterase, cholesterol oxidase and generates hydrogen peroxide, colour generation is reacted through Trinder ' s, by its absorbance measurement
SD-LDLC content out.
Stabilization of kit of the present invention is good, and accuracy is high, easy to operate quick, is suitable for automatic clinical chemistry analyzer and detects.
Detailed description of the invention
Fig. 1 is the test that the embodiment of the present invention 2 is measured in 7180 automatic clinical chemistry analyzer of Hitachi according to the measuring method
Result schematic diagram;
Fig. 2 is the test result schematic diagram that the embodiment of the present invention 2 and listing contrast agents measure 20 parts of samples simultaneously.
Specific embodiment
The present invention is described further With reference to embodiment:
Embodiment 1
Reagent R1:
Reagent R2:
Embodiment 2
Reagent R1:
Reagent R2:
Embodiment 3
Reagent R1:
Reagent R2:
Embodiment 4:
Reagent R1:
Reagent R2:
Measuring method
Reagent R1 and reagent R2 are placed in fully-automatic analyzer (Hitachi 7180) corresponding reagent position, draw 240 μ l it
R1 reagent, is added 3 μ l titers or sample stirs and evenly mixs in reaction cup, and 37 DEG C measure absorbance (A1) after five minutes, is added 80
μ l R2 reagent calculates SD- by A2-A1=Δ A with 550nm~600nm wavelength measurement absorbance (A2) after after five minutes
LDLC content.
Range of linearity verification test
With high level 2.6mM SD-LDLC sample, with physiological saline doubling dilution, with the embodiment of the present invention 2 in Hitachi 7180
Automatic clinical chemistry analyzer is measured according to the measuring method, result such as Fig. 1:
Correlation test
20 parts of samples are measured simultaneously with reagent embodiment 2 of the present invention and listing contrast agents, measuring method is same as above,
As a result following (see the table below and Fig. 2):
Reagent of the present invention
Claims (10)
1. a kind of sdLDL-C kit, it is characterised in that including independent reagent R1 and reagent R2,
It wherein include buffer, lipoprotein lipase, cholesterol esterase, cholesterol oxidase, heparin sodium, chlorination in reagent R1
Magnesium, seralbumin, nonionic surfactant, catalase or peroxidase, preservative;
The reagent R2 includes buffer, nonionic surfactant, Trinder ' s reactant B, preservative.
2. a kind of sdLDL-C kit according to claim 1, it is characterised in that reagent R1
In further include having Trinder ' s reactant A.
3. a kind of sdLDL-C kit according to claim 2, it is characterised in that further include
There is stabilizer.
4. a kind of sdLDL-C kit according to claim 1, it is characterised in that reagent R1
The concentration of middle buffer is 20~200mM, pH=6.0~7.5;The content of lipoprotein lipase be 0.1~2u/ml, cholesterol
The content of esterase is 0.5~2.0u/ml, the content of cholesterol oxidase is 0.3~1.5u/ml, the content of heparin sodium be 0.1~
0.5g/L, the concentration of magnesium chloride is 2~20mM, sero-abluminous content is 1~20g/L, nonionic surfactant contains
Amount be 0.1~2.0ml/L, catalase content be 50~200u/ml or peroxidase content be 0.3~3ku/L,
Trinder ' s reactant A is 1~4mM, the content of preservative is 0.005-0.105%.
5. a kind of sdLDL-C kit according to claim 1 or 4, it is characterised in that described
Buffer is Good ' s buffer, BES buffer, Bis-Tris buffer, Pipes buffer, Mops buffer, HEPES slow
One of fliud flushing.
6. a kind of sdLDL-C kit according to claim 1 or 4, it is characterised in that described
Nonionic surfactant is one or both of styrene polyoxyethylene ether, benzyl phenyl polyoxyethylene ether.
7. a kind of sdLDL-C kit according to claim 1, it is characterised in that the examination
The concentration of buffer is 20~200mM, pH=6.0~7.5 in agent R2;The mass concentration of the nonionic surfactant is
0.5~5%;The concentration of Trinder ' s reactant B is 2-5mM, the content of preservative is 0.005-0.105%.
8. a kind of sdLDL-C kit according to claim 1, it is characterised in that reagent R2
Described in buffer be Good ' s buffer, BES buffer, Bis-Tris buffer, Pipes buffer, Mops buffer,
One of HEPES buffer solution.
9. a kind of sdLDL-C kit according to claim 1, it is characterised in that reagent R2
Described in nonionic surfactant be Pluronic F68 or alkyl phenol polyoxyethylene ether.
10. a kind of sdLDL-C kit according to claim 1, it is characterised in that reagent R2
In further include weight percent be 0.1-0.2% Sodium azide.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201810034866.2A CN108841918A (en) | 2018-01-15 | 2018-01-15 | A kind of sdLDL-C kit |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201810034866.2A CN108841918A (en) | 2018-01-15 | 2018-01-15 | A kind of sdLDL-C kit |
Publications (1)
Publication Number | Publication Date |
---|---|
CN108841918A true CN108841918A (en) | 2018-11-20 |
Family
ID=64211736
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201810034866.2A Pending CN108841918A (en) | 2018-01-15 | 2018-01-15 | A kind of sdLDL-C kit |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN108841918A (en) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110791549A (en) * | 2019-12-03 | 2020-02-14 | 宁波普瑞柏生物技术股份有限公司 | Method and kit for quantitative determination of small dense low density lipoprotein cholesterol |
CN111398607A (en) * | 2019-01-03 | 2020-07-10 | 重庆中元汇吉生物技术有限公司 | Small and dense low-density lipoprotein cholesterol determination reagent |
CN113308513A (en) * | 2021-05-27 | 2021-08-27 | 宁波瑞源生物科技有限公司 | Small and dense low-density lipoprotein cholesterol detection kit and detection method thereof |
CN114487420A (en) * | 2022-01-14 | 2022-05-13 | 广西康柏莱科技有限公司 | Creatine kinase isoenzyme detection kit |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103589776A (en) * | 2013-09-12 | 2014-02-19 | 绍兴圣康生物科技有限公司 | Small and dense low-density lipoprotein cholesterol determination kit |
CN104673879A (en) * | 2015-03-07 | 2015-06-03 | 王贤俊 | Small and dense low-density lipoprotein cholesterin detection kit and preparation thereof |
CN105652021A (en) * | 2016-03-11 | 2016-06-08 | 上海练佰生物技术中心 | Reagent, method and kit for measuring small-and-dense lipoprotein |
CN107446989A (en) * | 2017-06-23 | 2017-12-08 | 美康生物科技股份有限公司 | Determine the method and reagent of the sdLDL-C in sample |
-
2018
- 2018-01-15 CN CN201810034866.2A patent/CN108841918A/en active Pending
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103589776A (en) * | 2013-09-12 | 2014-02-19 | 绍兴圣康生物科技有限公司 | Small and dense low-density lipoprotein cholesterol determination kit |
CN104673879A (en) * | 2015-03-07 | 2015-06-03 | 王贤俊 | Small and dense low-density lipoprotein cholesterin detection kit and preparation thereof |
CN105652021A (en) * | 2016-03-11 | 2016-06-08 | 上海练佰生物技术中心 | Reagent, method and kit for measuring small-and-dense lipoprotein |
CN107446989A (en) * | 2017-06-23 | 2017-12-08 | 美康生物科技股份有限公司 | Determine the method and reagent of the sdLDL-C in sample |
Non-Patent Citations (3)
Title |
---|
刘文演: "《生物化学》", 30 September 2009, 第二军医大学出版社 * |
李玉中: "《临床医学检验学 高级医师进阶》", 31 January 2016, 中国协和医科大学出版社 * |
陈三斌: "《农用化工产品手册》", 30 April 2008, 金盾出版社 * |
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111398607A (en) * | 2019-01-03 | 2020-07-10 | 重庆中元汇吉生物技术有限公司 | Small and dense low-density lipoprotein cholesterol determination reagent |
CN111398607B (en) * | 2019-01-03 | 2024-02-23 | 中元汇吉生物技术股份有限公司 | Small dense low density lipoprotein cholesterol determination reagent |
CN110791549A (en) * | 2019-12-03 | 2020-02-14 | 宁波普瑞柏生物技术股份有限公司 | Method and kit for quantitative determination of small dense low density lipoprotein cholesterol |
CN110791549B (en) * | 2019-12-03 | 2023-09-26 | 宁波普瑞柏生物技术股份有限公司 | Method and kit for quantitatively determining small dense low density lipoprotein cholesterol |
CN113308513A (en) * | 2021-05-27 | 2021-08-27 | 宁波瑞源生物科技有限公司 | Small and dense low-density lipoprotein cholesterol detection kit and detection method thereof |
CN113308513B (en) * | 2021-05-27 | 2022-02-18 | 宁波瑞源生物科技有限公司 | Small and dense low-density lipoprotein cholesterol detection kit and detection method thereof |
CN114487420A (en) * | 2022-01-14 | 2022-05-13 | 广西康柏莱科技有限公司 | Creatine kinase isoenzyme detection kit |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN108841918A (en) | A kind of sdLDL-C kit | |
CN103952462B (en) | The method and kit of quantification of small, dense LDL cholesterol | |
US8697378B2 (en) | Method and kit for quantitative determination for small, dense particle low density lipoproteins | |
DK2116612T3 (en) | REAGENT FOR DETERMINING QUANTITY OF LOW DENSITY LOW DENSITY LIPOPROTEIN | |
ES2335658T3 (en) | METHOD AND REAGENT THAT ALLOWS MEASURING CHOLESTEROL IN THE HIGH DENSITY PROTEIN LIPO. | |
US7811779B2 (en) | Method of multiquantification for cholesterol of low-density lipoprotein | |
EP2319937B1 (en) | Blood component measurement method utilizing hemolyzed whole blood, and kit for the method | |
CN107287277B (en) | Small and dense low-density lipoprotein cholesterol detection kit | |
JP4690200B2 (en) | Method, reagent and kit for determination of cholesterol in very low density lipoprotein remnant (VLDL remnant) | |
JP5299960B2 (en) | Method for diagnosing fatty liver disease, diagnostic device, diagnostic program, diagnostic agent and screening method for therapeutic agent for fatty liver disease | |
US20150132834A1 (en) | Method for removal of triglycerides in lipoproteins other than low-density lipoproteins | |
JP4456715B2 (en) | Method and reagent for measuring cholesterol in remnant-like lipoprotein | |
BR112014025875B1 (en) | MEASUREMENT METHOD | |
WO2017204230A1 (en) | Method and reagent for quantifying cholesterol in triglyceride-rich lipoprotein | |
CN112695071B (en) | High-density lipoprotein 3 determination reagent, method and kit | |
EP2634261A1 (en) | Method and kit for measuring cholesterol in low density lipoproteins | |
EP3026120B1 (en) | Method and reagent for quantifying cholesterol in high density lipoprotein 3 | |
JP4519239B2 (en) | Method for quantifying cholesterol in low density lipoprotein | |
JP2006180707A (en) | Method for determining content of triglyceride in low-density lipoprotein | |
US20200200769A1 (en) | Method for measuring glycated hemoglobin | |
JP3653564B2 (en) | Stable measuring reagent and measuring method | |
JP4865880B2 (en) | Method for quantifying cholesterol in low density lipoprotein | |
JPH09288111A (en) | Reagent for removing turbidity of biological sample |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination |