CN109580504B - Lipoprotein cholesterol determination reagent and kit - Google Patents
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Abstract
The invention provides a lipoprotein cholesterol determination reagent and a kit, belonging to the technical field of biochemical detection, and comprising a poly-alpha-olefin, cholesterol oxidase, a reaction promoter, cholesterol esterase, peroxidase, catalase, a catalase inhibitor, a surfactant, an emulsifier, polyanion, divalent cation, a stabilizer, an anti-interference agent, a color developing agent and a preservative. The invention can be used for the detection of a full-automatic biochemical analyzer, has simple and convenient operation, can improve the detection efficiency and accuracy and reduce the detection cost.
Description
Technical Field
The invention belongs to the technical field of biochemical detection, and particularly relates to a lipoprotein cholesterol determination reagent and a kit.
Background
According to the physical properties, the human plasma lipoproteins are mainly classified into four types: high Density Lipoproteins (HDL), Low Density Lipoproteins (LDL), Very Low Density Lipoproteins (VLDL) and Chylomicrons (CM), which are the main presence and transport forms of cholesterol in plasma. Wherein, the contents of high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) are respectively in negative and positive correlation with the incidence rate of human arteriosclerosis coronary heart disease, the contents of the HDL-C and the low density lipoprotein cholesterol (LDL-C) in human plasma are influenced by various factors, such as age, sex, genetic factors, smoking/exercise conditions, daily diet, obesity and the like, according to the judgment standard provided by the dyslipidemia prevention and treatment suggestions in China, and according to the judgment standard provided by the dyslipidemia prevention and treatment suggestions in China, under the normal condition, the HDL-C of a male is between 1.16 and 1.42mmol/L, and the HDL-C of a female is between 1.29 and 1.55 mmol/L; the average LDL-C content of young people is about 2.7mmol/L, and that of middle aged and elderly people is about 3.37 mmol/L. The determination of the HDL-C, LDL-C content has important significance and is an important risk factor index of cardiovascular and cerebrovascular diseases in clinical practice at present.
There are various methods for measuring the HDL-C, LDL-C content, and the methods for measuring HDL-C, LDL-C content are commercially available as direct measurement methods after ultracentrifugation and electrophoresis. The direct measurement method of HDL-C, LDL-C can be divided into three generations, and currently, the method is widely used in various clinical laboratories as a third-generation homogeneous measurement method, and the method is a two-step method, does not need human treatment such as sample pretreatment, precipitation separation and the like, has small reagent and sample dosage, can be used for measurement on a full-automatic biochemical analyzer, and has higher accuracy and precision of measurement results. The HDL-C homogeneous assay mainly comprises the following 5 types: PEG/antibody coating method (IRC), PEG enzyme modification method (PEGME), reaction promoter/peroxidase scavenging method (SPD), catalase scavenging method (CAT), and antibody immunoseparation method (AB). LDL-C homogeneous assay mainly comprises: soluble reaction method (SOL), surfactant scavenging method (SUR), protective reagent method (PRO), catalase scavenging method (CAT), calixarene method (CAL).
Different determination principles are different, but certain defects exist generally, Non-HDL-C or Non-LDL-C cannot be completely wrapped or removed in the first step of reaction, and the determination result is high easily; in the second step of reaction, HDL-C or LDL-C can not be completely released, which easily results in low measurement result; in a patient sample with abnormal phospholipid and cholesterol in plasma, the measurement of HDL-C or LDL-C is abnormal, and the anti-interference capability and the stability are poor.
Disclosure of Invention
The invention aims to provide a lipoprotein cholesterol determination reagent and a kit, which can be used for detection of a full-automatic biochemical analyzer, are simple and convenient to operate, can improve the detection efficiency and accuracy and reduce the detection cost.
The invention provides the following technical scheme:
a lipoprotein cholesterol measuring reagent contains poly-alpha-olefin, cholesterol oxidase and reaction accelerator.
Preferably, the concentration of the alpha-olefin is from 0.0005 to 0.005% (v/v).
Preferably, the molecular weight of the cholesterol oxidase is 25-45KDa, and the concentration is 0.1-2 KU/L.
Preferably, the reaction accelerator is at least one of flufenamic acid, mefenamic acid, fusidic acid, 6-dibromo-2, 2:6, 2-terpyridine and monensin, and the concentration of the reaction accelerator is 50-200 umol/L.
Preferably, the composition also comprises cholesterol esterase, peroxidase, catalase inhibitor, surfactant, emulsifier, polyanion, divalent cation, stabilizer, anti-interference agent, color developing agent and preservative.
Preferably, the cholesterol esterase is at least one of a modified enzyme or an unmodified enzyme, and the concentration is 0.1-2 KU/L.
Preferably, the enzyme modifier is at least one of PEG6000, double-arm PEG7000 and diethylenetriamine pentaacetic acid.
Preferably, the concentration of peroxidase is 0.1 to 3 KU/L.
Preferably, the concentration of catalase is 0.1 to 3 KU/L.
Preferably, the catalase inhibitor is one or two of sodium azide and 3-amino-1, 2, 4-triazole, and the concentration is 1-30 g/L.
Preferably, the surfactant is at least one of Emulgen B-66, Triton-X, trimethyl-beta-cyclodextrin, ethylene oxide octadecamine, poloxamer-F88, Bridgman-58, Tween series, SDS, ether carboxylate, alkyl glycerol ether, polyoxyethylene cetyl ether and polyoxyethylene octyl phenyl ether, and the HLB value of the surfactant is 10-15.
Preferably, the emulsifier is at least one of ethanol, fatty alcohol-polyoxyethylene ether and polyoxypropylene ether.
Preferably, the polyanion is at least one of heparin sulfate, dextran sulfate, sodium tungsten phosphate and polyethylene glycol, and the concentration is 0.1-1 g/L.
Preferably, the divalent cation is Mg2+、Mn2+、Ca2+At least one of them, the concentration is 0.1-5 g/L.
Preferably, the stabilizer can be at least one of glutathione, trehalose, lecithin, glycerol, EDTA, betaine, gelatin and bovine serum albumin, and the concentration is 0.1-5 g/L.
Preferably, the anti-interference agent is at least one of chloroform, protamine sulfate, mannitol and ascorbic acid oxidase, and the concentration is 0.1-6 g/L.
Preferably, the developer is at least one of phenol, sodium 3, 5-dichloro-2-hydroxybenzenesulfonate (DHBS), N-ethyl-N- (2-hydroxy-3-propanesulfo) m-Toluidine (TOOS), disodium N, N-bis (4-sulfobutyl) -3-methylaniline (TODB), and sodium N-ethyl-N- (3-sulfopropyl) -3-methylaniline (TOPS) at a concentration of 5-15 mL/L.
Preferably, the preservative is at least one of sodium azide, gentamicin, Proclin series and KroVin series, and the concentration is 1-15 g/L.
A kit for HDL-C determination comprises a first reagent and a second reagent, wherein the first reagent contains buffer solution, cholesterol oxidase, peroxidase, surfactant, anti-interference agent, reaction promoter, preservative, divalent cation, stabilizer and polyanion, and the second reagent contains buffer solution, catalase, surfactant, anti-interference agent, poly-alpha-olefin, preservative, stabilizer, color developing agent and 4-aminoantipyrine.
A kit for LDL-C assay comprising a first reagent comprising a buffer, a poly a-olefin, cholesterol oxidase, cholesterol esterase, catalase, surfactant, anti-interference agent, reaction promoter, preservative and stabilizer and a second reagent comprising a buffer, developer, 4-aminoantipyrine, peroxidase, surfactant, preservative, catalase inhibitor and stabilizer.
The invention has the beneficial effects that:
(1) the lipoprotein cholesterol determination reagent provided by the invention introduces poly-alpha-olefin to improve the accuracy of the determination of the lipoprotein cholesterol, and the poly-alpha-olefin can improve the selectivity of the surfactant.
(2) The lipoprotein cholesterol determination reagent provided by the invention introduces cholesterol oxidase with low molecular weight, can improve the clearance capability of lipoprotein cholesterol, further improve the accuracy of determination of different lipoprotein cholesterol, and the cholesterol oxidase can improve the oxidation of cholesterol in reaction.
(3) The lipoprotein cholesterol determination reagent provided by the invention introduces one or more than two reaction promoters to improve the determination efficiency of lipoprotein cholesterol, on one hand, the dosage of enzyme can be reduced, the cost is saved, and on the other hand, the reaction time can be shortened to a certain extent.
(4) The poly-alpha-olefin added into the kit for measuring HDL-C provided by the invention improves the specificity of the surfactant to HDL-C and improves the accuracy of HDL-C measurement.
(5) The added poly-alpha-olefin improves the selection of the surfactant for the LDL-C and improves the accuracy of the LDL-C measurement.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the principles of the invention and not to limit the invention. In the drawings:
FIG. 1 is a graph of the linear dependence of the test values and theoretical values of the kit of example 2 in test 1 of example 2;
FIG. 2 is a graph of the linear correlation of the example 2 kit and the control kit in test 4 of example 2;
FIG. 3 is a graph of the linear dependence of the test values and theoretical values of the example 3 kit of example 3 in test 1 of example 3;
FIG. 4 is a graph of the linear correlation of the example 3 kit and the control kit in test 4 of example 3.
Detailed Description
Example 1
A lipoprotein cholesterol measuring reagent contains poly-alpha-olefin, cholesterol oxidase and reaction accelerator. The concentration of the alpha-olefin is 0.0005-0.005% (v/v). The molecular weight of the cholesterol oxidase is 25-45KDa, and the concentration is 0.1-2 KU/L. The reaction accelerator is at least one of flufenamic acid, mefenamic acid, fusidic acid, 6-dibromo-2, 2:6, 2-terpyridine and monensin, and the concentration of the reaction accelerator is 50-200 umol/L.
The lipoprotein cholesterol determination reagent also comprises cholesterol esterase, peroxidase, catalase, a catalase inhibitor, a surfactant, an emulsifier, polyanion, divalent cation, a stabilizer, an anti-interference agent, a color developing agent and a preservative.
The cholesterol esterase (CHE) is at least one of modified enzyme and unmodified enzyme, and has a concentration of 0.1-2 KU/L. The enzyme modifier is at least one of PEG6000, double-arm PEG7000 and diethylenetriamine pentaacetic acid.
The Peroxidase (POD) can be derived from at least one of plant (such as semen glycines, radix Raphani), animal (such as animal leukocyte), and microorganism (such as yeast) at a concentration of 0.1-3 KU/L.
The Catalase (CAT) can be derived from at least one of plant (chloroplast), animal (such as bovine liver), and microorganism (such as Rhodopseudomonas) at a concentration of 0.1-3 KU/L.
The catalase inhibitor is one or two of sodium azide and 3-amino-1, 2, 4-triazole, and the concentration is 1-30 g/L.
The surfactant is at least one of Emulgen B-66, Triton-X, trimethyl-beta-cyclodextrin, ethylene oxide octadecamine, poloxamer-F88, Brij-58, Tween series, SDS, ether carboxylate, alkyl glycerol ether, polyoxyethylene cetyl ether and polyoxyethylene octyl phenyl ether, and the HLB value of the surfactant is 10-15.
The emulsifier is at least one of ethanol, fatty alcohol-polyoxyethylene ether and polyoxypropylene ether.
The polyanion is at least one of heparin sulfate, dextran sulfate, sodium tungsten phosphate and polyethylene glycol, and the concentration is 0.1-1 g/L.
The divalent cation being Mg2+、Mn2+、Ca2+At least one of them, the concentration is 0.1-5 g/L.
The stabilizer can be at least one of glutathione, trehalose, lecithin, glycerol, EDTA, betaine, gelatin, and bovine serum albumin, and has a concentration of 0.1-5 g/L.
The anti-interference agent is at least one of chloroform, protamine sulfate, mannitol and ascorbic acid oxidase, and the concentration is 0.1-6 g/L.
The developer is at least one of phenol, 3, 5-dichloro-2-hydroxy benzene sodium sulfonate (DHBS), N-ethyl-N- (2-hydroxy-3-propyl sulfo) m-Toluidine (TOOS), N-di (4-sulfobutyl) -3-methylaniline disodium (TODB) and N-ethyl-N- (3-sulfopropyl) -3-methylaniline sodium salt (TOPS), and the concentration is 5-15 mL/L.
The antiseptic is at least one of sodium azide, gentamicin, Proclin series, and KroVin series, and has a concentration of 1-15 g/L.
Example 2
A kit for HDL-C determination comprises a first reagent and a second reagent, wherein the first reagent contains buffer solution, cholesterol oxidase, peroxidase, surfactant, anti-interference agent, reaction promoter, preservative, divalent cation, stabilizer and polyanion, and the second reagent contains buffer solution, catalase, surfactant, anti-interference agent, poly-alpha-olefin, preservative, stabilizer, color developing agent and 4-aminoantipyrine.
The kit comprises the following specific raw materials in parts by weight:
TABLE 1 composition and concentration of the first reagent
| Buffer PIPES (PH 7.0) | 50mmol/L |
| Cholesterol oxidase (25-45KDa, CHO) | 1.5KU/L |
| Cholesterol esterase (CHE) | 1.5KU/L |
| Peroxidase (POD) | 1.5KU/L |
| Brij-58 | 0.5％ |
| Octadecylamine oxirane | 0.5％ |
| Protamine sulfate | 3g/L |
| Ascorbic acid oxidase (ASO) | 2KU/L |
| Flufenamic acid | 120umol/L |
| Proclin 300 | 5g/L |
| MgSO4 | 2g/L |
| Glycerol | 8ml/L |
| Dextran Sulfate (DS) | 0.5g/L |
| Alpha-cyclodextrin | 0.5g/L |
TABLE 2 composition and concentration of the second reagent
| Buffer PIPES (PH 7.0) | 50mmol/L |
| Catalase (CAT) | 1.5KU/L |
| Emulgen B-66 | 1％ |
| Mannitol | 5g/L |
| Poly alpha-olefins | 0.001％ |
| Proclin 300 | 5g/L |
| Betaine | 0.3mol/L |
| TOOS | 8mL/L |
| 4-aminoantipyrine (4-AAP) | 1g/L |
The SPD method in the homogeneous assay method is selected in this example, and is based on the following principle: HDL-C is directly determined by utilizing the affinity difference between the surfactant and different lipoproteins. The method mainly comprises two steps of reactions: in the first step, a reaction promoter (synthetic polymer/surfactant) is used to enable non-HDL to form a soluble complex, and free cholesterol on the surface of the complex is removed under the action of POD; in the second step of reaction, HDL particles are made soluble under the action of surfactant, and the released cholesterol and aminoantipyrine generate bluish purple benzoquinone pigment under the action of CHO, CHE and CAT. The aim of directly measuring HDL-C without precipitation separation is realized by measuring absorbance.
In this embodiment, HDL-C in a sample is detected, and the sample to be detected is one of serum and plasma, and the specific steps are as follows:
(1) uniformly mixing 300uL of first reagent and 3uL of sample to be detected, and incubating at 37 ℃ to obtain reaction liquid 1;
(2) then adding 100uL of a second reagent, uniformly mixing to obtain a reaction solution 2, and measuring the absorbance A1 of the reaction solution 2;
(3) measuring the absorbance A2 of the reaction solution 2 again after 5min, wherein the main wavelength is 600nm and the sub-wavelength is 700 nm;
(4) measuring the absorbance of the high-density lipoprotein cholesterol standard solution by the operations of the steps (1) to (3) to respectively obtain the absorbance A3 and the absorbance A4, wherein the main wavelength is 600nm, and the auxiliary wavelength is 700 nm;
(5) calculating by using the absorbance difference, and calculating the HDL-C content according to a formula: HDL-C concentration ═ sample absorbance (a2-a1) × standard solution concentration/standard solution absorbance (a 4-A3).
Examples of the experiments
HDL-C was tested simultaneously with the kit for HDL-C determination provided in this example using the same type of product from other companies as a control kit, and the results are shown in Table 3:
table 3 comparison of kit parameters with control kit parameters in example 2
| Linear Range (mmol/L) | Accuracy of | Analytical sensitivity (mmol/L) | |
| Control kit | 0.00～3.00 | ≦±10％ | 3.0 |
| EXAMPLE 2 kit | 0.00～5.50 | ≦±10％ | 1.5 |
The contrast kit is a domestic commercial HDL-C detection kit, and comprises the following components in parts by weight: the reagent 1 comprises PIPES buffer solution, TOOS, ascorbic acid oxidase and Tween 80; reagent 2 includes PIPES buffer, 4-AAP, CHOD, CHER, POD. As is clear from Table 3, the assay sensitivity of the kit of example 2 was higher.
Standard samples of different concentrations of HDL-cholesterol were prepared, 0, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 5, 5.5mmol/L, and their concentrations were determined using the kit of example 2 and the control kit, respectively, and all the tests were repeated 3 times and averaged. The results are shown in Table 4:
TABLE 4 Linear Range comparison
As can be seen from table 4 and the results of fig. 1: when the sample concentration is ≧ 3.5mmol/L, the linear range is small, the error of the control kit in measurement becomes large, and the HDL-C concentration cannot be detected accurately, while the kit of example 2 is still within the normal error range, so that the linear range of the kit of the invention is wider.
The control kit and the kit of example 2 were used as materials, calibration was performed using Landau calibrator, measurements were performed using Roche high and low quality controls, all measurements were performed for 3 replicates, and the average was taken. The results are shown in Table 5:
TABLE 5 comparison of accuracy
| Target value | Control kit | EXAMPLE 2 kit |
| High value of 1.53mmol/L | 1.56 | 1.52 |
| Low value of 0.844mmol/L | 0.85 | 0.84 |
Table 5 the results show that: the test result of the kit of example 2 is closer to the target value and has higher accuracy than the control kit.
Internal precision: the measurement was repeated 10 times (n is 10) using high and low roche quality control samples, respectively, and the coefficient of variation (CV%) value was calculated. Specifically, as shown in Table 6:
TABLE 6 comparison of in-batch precision
From the results of the above table it can be seen that: the kit of example 2 was more accurate in batch than the control kit.
The HDL-C content of 40 clinical samples was determined by calibration with Landau calibrators, and the results shown in Table 7 and FIG. 2 show that the test results of the kit of example 2 correlate well with those obtained with the control reagent, and all tests were performed in 3 replicates.
TABLE 7 comparison of Linear dependence
Example 3
A kit for LDL-C assay comprising a first reagent comprising a buffer, a poly a-olefin, cholesterol oxidase, cholesterol esterase, catalase, surfactant, anti-interference agent, reaction promoter, preservative and stabilizer and a second reagent comprising a buffer, developer, 4-aminoantipyrine, peroxidase, surfactant, preservative, catalase inhibitor and stabilizer.
The kit comprises the following specific raw materials in parts by weight:
TABLE 8 composition and concentration of the first reagent
TABLE 9 composition and concentration of the second reagent
| Buffer MOPS (PH 7.0) | 50mmol/L |
| TOOS color developing agent | 10mL/L |
| 4-aminoantipyrine (4-AAP) | 1g/L |
| Peroxidase (POD) | 1KU/L |
| Triton-X | 1.2％ |
| Proclin 300 | 5g/L |
| Sodium azide | 2g/L |
| Betaine | 1.5g/L |
The CAT method in the homogeneous assay was used in this example, and the method was based on the following principle: LDL-C was directly measured by using the difference in affinity between surfactants and different lipoproteins. The method mainly comprises two steps of reactions: in the first step of reaction, a surfactant and an ionic buffer solution are utilized to specifically act on non-LDL, so that the cholesterol of the non-LDL is released, and the released cholesterol is eliminated under the action of CAT; in the second step of reaction, under the action of another surfactant, the cholesterol in LDL particles is released, and the released cholesterol and aminoantipyrine generate the bluish-purple benzoquinone pigment under the action of CHO, CHE and POD. The purpose of directly measuring LDL-C without precipitation separation is realized by measuring the absorbance.
In this embodiment, the LDL-C in the sample is detected, and the sample to be detected is one of serum and plasma, and the specific steps are as follows:
(1) uniformly mixing a 300uL reagent first reagent and a 3uL sample to be detected, and incubating at 37 ℃ to obtain a reaction solution 1;
(2) then adding 100uL of reagent and second reagent, uniformly mixing to obtain reaction liquid 2, and measuring the absorbance A1 of the reaction liquid 2;
(3) measuring the absorbance A2 of the reaction solution 2 again after 5min, and measuring the main wavelength to be 545nm and the auxiliary wavelength to be 700 nm;
(4) measuring the absorbance of the high-density lipoprotein cholesterol standard solution by the operations of the steps (1) to (3) to respectively obtain the absorbance A3 and the absorbance A4, and measuring the main wavelength to be 545nm and the auxiliary wavelength to be 700 nm;
(5) calculating by using the absorbance difference, and calculating the LDL-C content according to the formula: LDL-C concentration is sample absorbance (a2-a1) × standard concentration/standard absorbance (a 4-A3).
Examples of the experiments
The test for LDL-C was performed simultaneously with the kit for LDL-C assay provided in this example, using the same type of product from other companies as the control kit, and the results are shown in Table 10:
TABLE 10 comparison of kit parameters for example 3 and control kits
| Linear Range (mmol/L) | Accuracy of | Analytical sensitivity (mmol/L) | |
| Control kit | 0.00～6.00 | ≦±10％ | 4.0 |
| Example 3 kit | 0.00～16.00 | ≦±10％ | 2 |
The contrast kit is a domestic LDL-C detection kit, and comprises the following components in parts by weight: reagent 1 comprises Good's buffer; reagent 2 includes Good's buffer solution, 4-AAP, CHOD, CHER, POD. As is clear from Table 10, the reagent kit of example 3 has higher analytical sensitivity
Standard samples of different concentrations of ldl-cholesterol were prepared, 0, 2,4, 6, 8, 10, 12, 14, 16mmol/L, and their concentrations were determined using the example 3 kit and the control kit, respectively, and all tests were repeated 3 times and averaged. The results are shown in Table 11:
TABLE 11 Linear Range comparison
As can be seen from table 11 and the results of fig. 3: when the sample concentration is ≧ 6mmol/L, the linear range is small, the error of the control kit in measurement becomes large, and the concentration of LDL-C cannot be detected more accurately, while the kit of example 3 is still within the normal error range, so that the linear range of the kit of the invention is wider.
The control kit and the kit of example 3 were used as materials, calibration was performed using Landau calibrator, measurements were performed using Roche high and low quality controls, all measurements were performed for 3 replicates, and the average was taken. The results are shown in Table 12:
TABLE 12 accuracy comparison
| Target value | Control kit | Example 3 kit |
| High value 2.62mmol/L | 2.55 | 2.64 |
| Low value 1.56mmol/L | 1.43 | 1.53 |
The results show that: the test result of the kit of example 3 is closer to the target value and has higher accuracy than the control kit.
Internal precision: the measurement was repeated 10 times (n is 10) using high and low roche quality control samples, respectively, and the coefficient of variation (CV%) value was calculated. Specifically, as shown in table 13:
TABLE 13 comparison of in-batch precision
From the results of the above table it can be seen that: the kit of example 3 had higher in-batch precision than the control kit.
The LDL-C content of 40 clinical samples was determined by calibration with Landau calibrators, and the results of Table 14 and FIG. 4 below show that the test results of the kit of example 3 correlate well with those of the control reagent, and all tests were performed in 3 replicates and averaged.
TABLE 14 Linear correlation comparison
Although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (7)
1. A lipoprotein cholesterol assay reagent, characterized by containing a poly-alpha-olefin, a cholesterol oxidase and a reaction accelerator; the concentration of the poly-alpha-olefin is 0.0005 to 0.005% (v/v); the molecular weight of the cholesterol oxidase is 25-45KDa, and the concentration is 0.1-2 KU/L; the reaction accelerator is at least one of flufenamic acid, mefenamic acid, fusidic acid, 6-dibromo-2, 2:6, 2-terpyridine and monensin, and the concentration of the reaction accelerator is 50-200 umol/L.
2. The reagent for measuring lipoprotein cholesterol as claimed in claim 1, further comprising cholesterol esterase, peroxidase, catalase inhibitor, surfactant, emulsifier, polyanion, divalent cation, stabilizer, anti-interference agent, color-developing agent and preservative.
3. The reagent for measuring lipoprotein cholesterol as claimed in claim 2, wherein the surfactant is at least one of Emulgen B-66, Triton-X, trimethyl- β -cyclodextrin, ethyleneoxide octadecylamine, poloxamer-F88, brij-58, tween, SDS, ether carboxylate, alkyl glyceryl ether, polyoxyethylene cetyl ether, polyoxyethylene octyl phenyl ether, and the surfactant has HLB value of 10-15.
4. The reagent for assaying lipoprotein cholesterol according to claim 2, wherein the divalent cation is Mg2+、Mn2+、Ca2+At least one of them, the concentration is 0.1-5 g/L.
5. The reagent for measuring lipoprotein cholesterol as claimed in claim 2, wherein the stabilizer is at least one of glutathione, trehalose, lecithin, glycerol, EDTA, betaine, gelatin, bovine serum albumin, and has a concentration of 0.1-5 g/L.
6. A kit for HDL-C determination, comprising a first reagent and a second reagent, wherein the first reagent comprises a buffer solution, cholesterol oxidase, peroxidase, a surfactant, an anti-interference agent, a reaction promoter, a preservative, a divalent cation, a stabilizer and a polyanion, the cholesterol oxidase has a molecular weight of 25-45KDa and a concentration of 0.1-2 KU/L; the second reagent contains buffer solution, catalase, surfactant, anti-interference agent, poly-alpha-olefin, preservative, stabilizer, color developing agent and 4-aminoantipyrine; the concentration of the poly-alpha-olefin is 0.0005-0.005% (v/v).
7. A kit for LDL-C assay comprising a first reagent and a second reagent, said first reagent comprising a buffer, a poly a-olefin, a cholesterol oxidase, a cholesterol esterase, a catalase, a surfactant, an anti-interference agent, a reaction promoter, a preservative and a stabilizer, said cholesterol oxidase having a molecular weight of 25-45KDa and a concentration of 0.1-2KU/L, said poly a-olefin having a concentration of 0.0005-0.005% (v/v); the second reagent contains buffer solution, color developing agent, 4-aminoantipyrine, peroxidase, surfactant, preservative, catalase inhibitor and stabilizer.
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| US9084745B2 (en) * | 2008-05-22 | 2015-07-21 | H. Lundback A/S | Modulation of the Vps10p-domain for the treatment of cardiovascular disease |
| WO2014207087A1 (en) * | 2013-06-26 | 2014-12-31 | Abengoa Bioenergia Nuevas Tecnologias S.A. | Production of advanced fuels and of chemicals by yeasts on the basis of second generation feedstocks |
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| CN106383116B (en) * | 2016-10-21 | 2019-02-01 | 北京世纪沃德生物科技有限公司 | A kind of kit detecting high-density lipoprotein cholesterol |
| CN108627510A (en) * | 2018-06-06 | 2018-10-09 | 临安卡尔生物技术有限公司 | High-density lipoprotein cholesterol detection kit |
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