119 results about "Cholesterol Esterase" patented technology

Reagents, assays, methods, kits, devices, and systems for rapid measurement of cholesterol and cholesterol sub-fractions from a blood sample are provided. Total cholesterol, low density lipoprotein cholesterol, and high density lipoprotein cholesterol can be measured in a single assay using kinetic measurements, under conditions in which cholesterol sub-species are converted to a detectable product at distinct rates. The detectable product is measured at different times after assay initiation. A lipase, cholesterol esterase, cholesterol oxidase and a peroxidase may be used together to produce colored product in amounts directly proportional to the quantity of cholesterol converted. Methods for calculating very-low density lipoprotein cholesterol levels by further including triglyceride measurements are disclosed. Assays may be performed in a single reaction mixture, allowing more accurate and precise cholesterol determinations, including ratios of cholesterol sub-fractions to total cholesterol, at less expense, than would be expected by performing several different assays in different reaction mixtures.

The invention discloses a reagent, a method and a kit for measuring small-and-dense lipoprotein. The reagent comprises first agents and second agents, the first agents include 1-500 mg/L reagent A, 10-300 U/mL sodium heparin, 0.1-90 mmol/L divalent metal ion, 0.5-2 KU/L cholesterol esterase, 1-3.5 KU/L cholesterol oxidase, 100-300 KU/L catalase, 0.1-10 mmol/L 4-amino antipyrine, 0.05-6% KU/L surfactant A, and the second reagents include 0.2-10 KU/L peroxidase, 0.3-20 mmol/L Trinder's chromogen compound, 0.01-0.3% sodium azide and 0.05-12% surfactant B. The reagent can be used for specifically detecting sdLDL. By using the method, sdLDL can be detected specifically. The kit is simple and convenient to operate.

The use of fatty acid oxidizing enzymes in the manufacture of paper materials, such as paper, linerboard, corrugated paperboard, tissue, towels, corrugated containers and boxes. Examples of fatty acid oxidizing enzymes are oxygenases classified as EC 1.13.11. including any of the sub-classes thereof, such as lipoxygenase, EC 1.13.11.12. The effect of these enzymes is that the deposition of pitch is reduced, and bleaching and de-inking effects are also observed on the paper pulp and the resulting paper material. The fatty acid oxidizing enzyme can be used in combination with a substrate, with proteases, lipases, xylanases, cutinases, oxidoreductases, cellulases, endoglucanases amylases, mannanases, steryl esterases, and/or cholesterol esterases; or with surfactants and other adjuvants.

There is provided a method for a selective assay of component, particularly cholesterol, in very low-density lipoprotein (VLDL) which is one of serum lipoproteins.

In the assay, an enzymatic reaction of lipoprotein lipase (LPL) or cholesterol esterase (CE) which well reacts with high-density lipoprotein (HDL) and VLDL is carried out at least in the presence of calixarene or a salt thereof. It is also carried out in the presence of one or more substance(s) selected from albumin and basic amino acids in addition to calixarene or a salt thereof.

The invention discloses a detection kit for LDL (low-density lipoprotein) cholesterol and a use method of the detection kit, relates to the field of biochemical detection and aims to provide a detection kit, having high stability, accuracy and precision as well as low toxicity, for LDL cholesterol and a use method of the detection kit. The kit comprises a reagent 1 and a reagent 2, wherein the reagent 1 is prepared from a surfactant, polyethylene glycol, SDS (sodium dodecyl sulfate), Emulgen A9 series, CHOD (cholesterol oxidase), CHER (cholesterol esterase), ascorbic acid oxidase, CAT (catalase), 4-AAP (4-aminoantipyrine) and the like; the reagent 2 is prepared from octylphenyl polyethylene glycol, cholate, a compound stabilizer, bovine serum albumin, POD (peroxidase), TOPS (sodium 3-(N-ethyl-3-methylanilino)propanesulfonate) and the like. The proclin series added to the kit is a novel biological preservative, has good compatibility with various enzymes, better stability and low toxicity, and the stability of the reagents is maintained.

Disclosed is a method for quantifying cholesterol in high-density lipoproteins in a single step assay using a dry slide. The method for quantifying cholesterol in high-density lipoprotein comprises a first step of adding a sample onto a multi-layered dry slide wherein at least one of the layers contains phosphotungstic acid and another contains a high-density lipoprotein selective surfactant. The phosphotungstic acid precipitates non-high-density lipoproteins while the high-density lipoprotein selective surfactant only solubilizes high-density lipoproteins and does not solubilize non-HDL precipitated complexes. The cholesterol esterase then reacts with the solubilized HDL cholesterol esters to form cholesterol. Finally the cholesterol in the high-density lipoprotein is detected and quantified.

The invention provides a small and dense low-density lipoprotein cholesterin (sdLDL-C) detection kit which comprises double liquid reagents, including a reagent R1 and a reagent R2, wherein the reagent R1 comprises an MOPS buffer liquid (the pH value is 7.0), cholesterol esterase, cholesterol oxidase, phospholipase, catalase, polyoxyethylene alkyl phenyl ether (JK-14) as a surfactant A, polyoxyethylene benzyl styrene ether (A3PK) as a protecting agent, and N-ethyl-N-(2-hydroxyl-3-sulfopropyl)-3-methylaniline sodium salt (TOOS); the reagent R2 comprises an MOPS buffer liquid (the pH value is 7.0), peroxidase, polyethylene glycol octylphenol ether (Triton X-100) as a surfactant, 4-amino antipyrine (4-AA), and sodium azide. The method provided by the invention is high in sensitivity, high in specificity, simple in reagent preparation, free of sample pretreatment, and worthy of popularization and application.

The invention discloses a total cholesterol detection reagent and total cholesterol detection paper. The total cholesterol detection reagent comprises 6-9 KU/L cholesterol esterase, 3-5 KU/L cholesterol oxidase, 15-20 KU/L horseradish peroxidase, 3-5 KU/L ascorbic acid oxidase, 0.20-0.35 g/L 4-ampyrone, and 0.20-0.30 g/L color-showing matter. The detection reagent contains a plurality of specific enzymes in a specific ratio, and can achieve the purposes of rapid detection and relatively accurate detection. The detection paper comprises a reaction base layer, a reaction layer, a blood filtering layer and a hydrophilic layer, which are overlapped to form a siphon system, thereby further ensuring rapid detection and relatively accurate detection.

The invention discloses a method and reagent for detecting small and dense low-density lipoprotein cholesterol (sdLDL-C) in a sample. The method is characterized by comprising the steps of (1) eliminating cholesterol, except the sdLDL-C, in lipoproteins in the sample first in the presence of cholesterol esterase and an ion selective agent; and (2) conducting quantitative detection on remaining sdLDL-C obtained after treatment in step (1). The ion selective agent is used to selectively eliminate lipoproteins other than the sdLDL-C, so as to detect the sdLDL-C.

A method for specifically quantifying HDL cholesterol in which cholesterol in lipoproteins other than HDL is erased in the first step, and HDL cholesterol is specifically quantified in the second step, by which accurate values can be obtained even in measurements of abnormal samples such as disorder of lipid metabolism and lipoprotein abnormality, is disclosed. The method for quantifying cholesterol in high density lipoprotein according to the present invention comprises a first step of erasing cholesterol in lipoproteins other than high density lipoprotein by treating a test sample with cholesterol esterase and cholesterol oxidase in the absence of a surfactant which acts on high density lipoprotein and removing generated hydrogen peroxide; and a second step of adding a surfactant which specifically acts on high density lipoprotein to the product of said first step and quantifying hydrogen peroxide generated from cholesterol in high density lipoprotein by actions of cholesterol esterase and cholesterol oxidase. As the cholesterol oxidase used in the first step, one having a molecular weight of not more than 60 kilodaltons is used.

The invention discloses a high-density lipoprotein cholesterol detection kit which consists of a first reagent and a second reagent, wherein the volume of the first reagent is 3 times of that of the second reagent; the kit comprises a surfactant, cholesterol esterase, cholesterol oxidase, peroxidase, absolute ethyl alcohol and a thiophosphoric reagent. In the first reagent, non-high-density lipoprotein cholesterol in a serum sample is in reaction with the cholesterol esterase, the cholesterol oxidase and the peroxidase under the action of the surfactant, thereby being eliminated; in the secondreagent, the rest serum with high-density lipoprotein cholesterol is extracted through the absolute ethyl alcohol, proteins in the serum are denaturated, the solubility is degraded, then precipitateis generated, cholesterol is dissolved in the ethanol, and under the action of the thiophosphoric reagent, the cholesterol is in reaction with the thiophosphoric reagent, and then the content of the cholesterol is detected. Test results show that the kit disclosed by the invention is wide in linear range, high in accuracy and precision and in addition good in interference resistance.

The invention discloses a high-performance small and dense low-density lipoprotein cholesterol detection kit. The kit comprises the following raw materials in parts by weight; a reagent A: 90 to 110 mmol/L of a Good's buffer solution; 1-3 ku/L of cholesterol esterase, 1-2 ku/L of cholesterol oxidase, 0.7-0.9 ku/L of phospholipase, 400-600 ku/L of catalase, and 1-3 mmol/L N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3-methylaniline sodium salt (TOOS); and a reagent B: 90-110 mmol/L of the Good's buffer solution, 2-3 ku/L of peroxidase, 3-5 mmol/L of 4-aminoantipyrine and 0.04-0.06% of sodium azide. Theinvention relates to the technical field of lipoprotein cholesterol detection, and discloses the high-performance small and dense low-density lipoprotein cholesterol detection kit. The non-sdLDL-C components are removed through cholesterol lipase, cholesterol oxidase, phospholipase and catalase; the interference of other lipoprotein cholesterol on the detection process is avoided, the capacity ofthe small and dense low-density lipoprotein cholesterol in serum can be directly determined by using a full-automatic biochemical analyzer, and the kit is suitable for the requirements of clinical andlaboratory on the determination of the content of the small and dense low-density lipoprotein cholesterol.

The invention discloses a kit for detecting small and dense low-density lipoprotein cholesterin. The kit is characterized by comprising polyoxyethylene fatty acid ester, cholesterol esterase, cholesterol oxidase, dodecyl polypeptide and triton X-100, wherein the small and dense low-density lipoprotein cholesterin in a serum sample is protected under the action of the polyoxyethylene fatty acid ester, reaction of the small and dense low-density lipoprotein cholesterin with the cholesterol esterase and the cholesterol oxidase is inhibited to a certain extent, and the other lipoprotein cholesterols react with the cholesterol esterase and the cholesterol oxidase under the action of the dodecyl polypeptide to be eliminated; and the triton X-100 reacts with the remaining small and dense low-density lipoprotein cholesterin to detect the content thereof. The kit can be used for quickly and conveniently detecting sdLDL-C in the serum sample. The reagent formula is simple and easy to prepare and low in cost, large-scale industrial preparation can be carried out, and meanwhile, the kit is capable of efficiently, accurately and specifically detecting the small and dense low-density lipoprotein cholesterin in the sample.

The invention provides milky tea with the function of reducing blood sugar and blood fat, which comprises 10-50% (weight) of milk, 0.3-10% (weight) of tea powder and 0.03-40% (weight) of combination of xylo-oligosaccharide and L-araba sugar. The milky tea uses the combination of xylo-oligosaccharide and L-araba sugar, has the effect of synergistically inhibiting activity of alpha-glucosidase to reduce blood sugar of the human bodies, and has the effect of aiming at lipase activity and cholesteryl ester activity to synergistically enhance inhibition functions to reduce blood fat. In addition, the milky tea uses the combination of xylo-oligosaccharide and L-arabinose, can restrain sediment and has the effect of a clarifying agent.

The invention discloses a reagent for lipid typing detection. The reagent is characterized by comprising alkylphenol polyethoxylate, Triton X-100, cholesterol esterase, cholesterol oxidase, peroxidase, phosphate buffer, phenol, 4-aminoantipyrine, and PC300. For a formula of the reagent, lipoprotein can be dissolved and release cholesterol or cholesteryl ester contained internally through additionof alkylphenol polyethoxylate and Triton X-100 in a specific ratio and in a mixing ratio, cholesterol ester can generate cholesterol under the action of cholesterol esterase, cholesterol can generatecholest-4-ene-3-ketone and hydrogen peroxide under the action of cholesterol oxidase, hydrogen peroxide acts with phenol and 4-aminoantipyrine (4-AAP) under the action of peroxidase so as to generatea colored material, and contents of different lipoproteins are calculated by checking changes of absorbance.

The invention discloses a cholesterol esterase. An amino acid sequence of the cholesterol esterase is an amino acid sequence shown in the formula of SEQ ID NO: 1, or is an amino acid sequence which is obtained by processes of deletion, addition and/or replacement of one or more amino acids on the amino acid sequence shown in the formula of SEQ ID NO: 1 and maintains original cholesterol esterase functions. The invention also discloses a nucleotide sequence for coding the cholesterol esterase, a recombinant vector of the nucleotide sequence, a recombinant host cell of the recombinant vector, amethod for purifying the cholesterol esterase from the recombinant host cell, and a kit containing at least one of the cholesterol esterase, the nucleotide sequence, the recombinant vector and the recombinant host cell. The cholesterol esterase has good heat resistance and high stability. The kit can carry out rapid, effective and accurate detection on genes to guarantee prompt case diagnosis andtreatment, accurate aetiology investigation, and establishment of scientific prevention and control policies.

A reagent for fractional measurement of small, dense LDLs without pretreatment of a specimen, which is adaptable to a rapid and convenient autoanalyzer, is provided. A method for quantitatively determining cholesterol in small, dense LDLs in a sample is further provided, comprising: a first step of eliminating lipoproteins other than small, dense LDLs in a sample in the presence of cholesterol esterase and 0.05 g/L to 1.0 g/L of a surfactant that acts on the lipoproteins other than small, dense LDLs; and a second step of quantitatively determining cholesterol in small, dense LDLs that remain after the first step.

The invention discloses a small dense low-density lipoprotein cholesterol kit. The kit consists of two independent reagents which are R1 and R2; the R1 contains a buffer solution, lipoprotein lipase,a nonionic surfactant, serum albumin, heparin sodium, magnesium ions, cholesteryl esterase, cholesterol oxidase, catalase or peroxidase, a Trinder's reactant A, a stabilizing agent and an antiseptic;and the R2 contains the buffer solution, the nonionic surfactant, a Trinder's reactant B, sodium azide and the antiseptic. The reagent R1 can be taken to mix with a specimen to eliminate cholesterol except for SD-LDLC; and reaction can be performed on the SD-LDLC after the reagent R2 is added, the Trinder's reactant B can be stained to determine absorbance, and content can be calculated. The kit is good in stability, high in accuracy, simple and fast in operation, and suitable for automatic biochemical analyzer detection.

The invention provides a lipoprotein cholesterol determination reagent and a kit, belonging to the technical field of biochemical detection. The reagent comprises poly-alpha-olefin, cholesterol oxidase and reaction promoter, and further comprises cholesterol esterase, peroxidase, catalase, catalase inhibitor, surface active agent, emulsifier, polyanion, divalent cation, stabilizer, anti-interference agent, color developing agent and preservative. The reagent and the kit provided by the invention can be used for full-automatic biochemical analyzer detection, are simple and convenient to operate, can improve detection efficiency and accuracy and can reduce detection cost.

The invention discloses test paper for measuring high density lipoprotein cholesterol and a preparation method and application thereof. The test paper comprises a red blood cell separation layer, an auxiliary separation layer and a reaction layer, wherein the red blood cell separation layer, the auxiliary separation layer and the reaction layer are prepared from a separation material, a separationmembrane and a reaction membrane through carrying out steeping by red blood cell separation solution, auxiliary separation solution and reaction solution and then carrying out drying; the red blood cell separation solution comprises the following components: buffer salt, a divalent metal ion, dextran sulfate sodium salt with a molecular weight of 30000-500000, hemagglutinin or an Anti-RBC antibody; the auxiliary separation solution comprises the following components: buffer salt, a surfactant I and polyhydric alcohol; and the reaction solution comprises the following components: buffer salt,a surfactant II, polysaccharide, cholesterol esterase, cholesterol oxidase, peroxidase, ascorbic acid oxidase, an oxidizing substance and a color developing agent. The test paper is capable of simplyand rapidly detecting the contents of high density lipoprotein cholesterol. The invention furthermore provides a preparation method and application of the test paper.

The invention discloses a detecting kit. The kit comprises two reagents, wherein the first reagent R1 comprises beta-cyclodextrin, an anti-human ApoB100 monoclonal antibody, an anti-human ApoAI monoclonal antibody, ascorbic acid oxidase, cholesterol esterase and a Tris buffer solution, and the second reagent R2 comprises a Tris bufer solution, peroxidase, cholesterol oxidase and 4-ampyrone. The invention also provides a method for detecting cholesterol concentration in a lipoprotein remnant, and the method comprises the following steps of: adding a sample into the first reagent R1, carrying out incubation at 37 DEG C for 5 min, reading and measuring the sample A1 at the first point using a biochemistry analyzer, adding the sample into the second reagent R1, carrying out incubation at 37 DEG C for 5 min, reading and measuring the sample A2 at the second point, and calculating the cholesterol concentration in the lipoprotein remnant according to a formula. The measurement method disclosed by the invention is simple, convenient and fast.

This invention provides a method for separately or simultaneously quantifying cholesterol in a total amount of HDL-C, cholesterol in an HDL subfraction of apoE-containing HDL-C, and cholesterol in an HDL subfraction of apoE-deficient HDL-C. The method comprises enzymatically and separately quantifying cholesterol in apoE-containing HDL and cholesterol in apoE-deficient HDL by adding a surfactant selected from the group consisting of a surfactant with an apoE-containing HDL response rate/apoE-deficient HDL response rate ratio of 0.7 to less than 1.3, a surfactant with an apoE-containing HDL response rate/apoE-deficient HDL response rate ratio of less than 0.7, and a surfactant with an apoE-containing HDL response rate/apoE-deficient HDL response rate ratio of 1.3 or more to a test sample, allowing cholesterol esterase and cholesterol oxidase to react therewith, and quantifying the hydrogen peroxide generated.