CN108841918A - A kind of sdLDL-C kit - Google Patents

A kind of sdLDL-C kit Download PDF

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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
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reagent
buffer
sdldl
content
cholesterol
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俞宏儒
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Zhongshan Biaojia Biotechnology Co Ltd
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Zhongshan Biaojia Biotechnology Co Ltd
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    • 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
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    • 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
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    • 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
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    • 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

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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

A kind of sdLDL-C kit
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.
CN201810034866.2A 2018-01-15 2018-01-15 A kind of sdLDL-C kit Pending CN108841918A (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
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

Citations (4)

* Cited by examiner, † Cited by third party
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

Patent Citations (4)

* Cited by examiner, † Cited by third party
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)

* Cited by examiner, † Cited by third party
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 (6)

* Cited by examiner, † Cited by third party
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

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