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Lipoprotein cholesterol determination reagent and kit

A cholesterol and lipoprotein technology, applied in the field of biochemical detection, can solve problems such as inability to wrap or remove, weak anti-interference ability, poor stability, etc., and achieve the effect of improving removal ability, selection, and accuracy

Active Publication Date: 2019-04-05
WHITMAN BIOTECH NANJING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Different assay methods have different principles, but they all generally have certain defects. In the first step reaction, Non-HDL-C or non-LDL-C cannot be completely wrapped or removed, which may easily lead to high measurement results; in the second step reaction HDL-C or LDL-C cannot be released completely, which can easily lead to low measurement results; in samples from patients with abnormal phospholipids and cholesterol in plasma, HDL-C or LDL-C will be abnormally measured, with weak anti-interference ability and poor stability

Method used

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  • Lipoprotein cholesterol determination reagent and kit
  • Lipoprotein cholesterol determination reagent and kit
  • Lipoprotein cholesterol determination reagent and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] A reagent for measuring lipoprotein cholesterol contains polyalkene, cholesterol oxidase and a reaction accelerator. The concentration of the α-olefin is 0.0005-0.005% (v / v). The molecular weight of the cholesterol oxidase is 25-45KDa, and the concentration is 0.1-2KU / L. The reaction accelerator is at least one of flufenamic acid, mefenamic acid, fusidic acid, 6,6-dibromo-2,2:6,2-terpyridine, and monensin. It is 50-200umol / L.

[0040] The lipoprotein cholesterol assay reagent also includes cholesterol esterase, peroxidase, catalase, catalase inhibitor, surfactant, emulsifier, polyanion, divalent cation, stabilizer, anti-interference agent , developer and preservatives.

[0041] Cholesterol esterase (CHE) is at least one of modified enzyme or unmodified enzyme, and the concentration is 0.1-2KU / L. The modification of the enzyme is at least one of PEG6000, double-arm PEG7000 and diethylenetriaminepentaacetic acid.

[0042] Peroxidase (POD) can be derived from at least...

Embodiment 2

[0054] A test kit for measuring HDL-C, comprising a first reagent and a second reagent, the first reagent contains buffer, cholesterol oxidase, peroxidase, surfactant, anti-interference agent, reaction accelerator , preservative, divalent cation, stabilizer and polyanion, the second reagent contains buffer, catalase, surfactant, anti-interference agent, poly-alkene, preservative, stabilizer, color development agent and 4-aminoantipyrine.

[0055] The specific raw materials and proportions of this kit are as follows:

[0056] The composition and concentration of the first reagent of table 1

[0057] 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

Bridger-58

0.5％

Ethylene oxide octadecylamine

0.5％

protamine sulfate

3g / L

Ascorbate Oxidase (ASO)

2KU / L

flufenamic acid

120umol / L

Proclin 300 ...

experiment example

[0068] Using the same type of products from other companies as a control kit, HDL-C was tested simultaneously with the kit for HDL-C determination provided in this example, and the results are shown in Table 3:

[0069] Table 3 embodiment 2 kit and contrast kit parameter comparison

[0070]

Linear range (mmol / L)

Accuracy

Analytical sensitivity (mmol / L)

control kit

0.00～3.00

≦±10％

3.0

Example 2 Kit

0.00～5.50

≦±10％

1.5

[0071] The control kit is a commercially available HDL-C detection kit in China. Its components and formula are as follows: Reagent 1 includes PIPES buffer, TOOS, ascorbate oxidase, Tween 80; Reagent 2 includes PIPES buffer, 4-AAP, CHOD, CHER, POD. As can be seen from Table 3, the analytical sensitivity of the kit in Example 2 is higher.

[0072] test 1

[0073] Prepare standard samples of different concentrations of high-density lipoprotein cholesterol, respectively 0, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3....

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Abstract

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.

Description

technical field [0001] The invention belongs to the technical field of biochemical detection, and in particular relates to a reagent and a kit for measuring lipoprotein cholesterol. Background technique [0002] According to different physical properties, human plasma lipoproteins can be mainly divided into the following four types: high-density lipoprotein (HDL), low-density lipoprotein (LDL), very low-density lipoprotein (VLDL) and chylomicrons (CM). It is the main form of existence and transport of cholesterol in plasma. Among them, the contents of high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) are respectively negatively and positively correlated with the incidence of human atherosclerosis and coronary heart disease. Affected by many factors, such as age, gender, genetic factors, smoking / exercise, daily diet, obesity, etc. Under normal circumstances, the HDL-C of men is between 1.16-1.42mmol / L, and that of women is between ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31
CPCG01N21/31
Inventor 常俊骏费云燕孙文勇梁芬郑佳
Owner WHITMAN BIOTECH NANJING
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