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Method for measuring cholesterol in high-density lipoprotein, and reagent for use in said method

A technology of high-density lipoprotein and assay method, applied to the field of cholesterol assay in high-density lipoprotein and the reagents used therein

Active Publication Date: 2016-04-13
SEKISUI MEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these practical methods and reagents for clinical examination also have problems in the measurement accuracy. Deviation of measured values ​​during reagent measurement has also been a problem internationally (Non-Patent Document 4)

Method used

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  • Method for measuring cholesterol in high-density lipoprotein, and reagent for use in said method
  • Method for measuring cholesterol in high-density lipoprotein, and reagent for use in said method
  • Method for measuring cholesterol in high-density lipoprotein, and reagent for use in said method

Examples

Experimental program
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Embodiment

[0115] The following examples are given to further describe the present invention in detail, and the present invention is not limited by these. In addition, in this Example and a comparative example, the following manufacturer's reagent and enzyme were used. In addition, the ingredient name of a surfactant describes the name described in the product manual of each manufacturer.

[0116] MES (Dojin Chemical Laboratory Co., Ltd.), sodium hydroxide (KISHIDA Chemical Co., Ltd.), TOOS (Dojin Chemical Laboratory Co., Ltd.), 4-aminoantipyrine (Wako Pure Chemical Industries, Ltd.), sodium phosphotungstate (KISHIDA Chemical Co., Ltd.), sodium dextran sulfate (molecular weight 4000) (Tokyo Chemical Industry Co., Ltd.), sodium dextran sulfate (molecular weight 36000-50,000) (Wako Pure Chemical Industries, Ltd.), sodium dextran sulfate ( Molecular weight 500,000) (Wako Pure Chemical Industry Co., Ltd.), magnesium chloride (KISHIDA Chemical Co., Ltd.), tributyl-n-octylphosphine chloride (...

Embodiment 1

[0136] (Example 1) Screening of Compounds Effective in Solving Problems

[0137]A sample obtained by using the HDL fraction and VLDL fraction prepared by ultracentrifugation and diluting them with physiological saline so that the cholesterol concentration in the two fractions is 50 mg / dL (hereinafter, this sample may be referred to as For the HDL fraction sample and the VLDL fraction sample), screening for a compound that lowers the reactivity of the enzyme for measuring cholesterol with the VLDL fraction was performed. The reagents used for screening were the basic first reagent described in Comparative Example 1, and polyanions, various Reagents obtained from salt compounds and BSA (Example 1-1 to Example 1-16). As the second reagent, the basic second reagent described in Comparative Example 1 was used.

[0138] [table 3]

[0139]

[0140] However, in the table, PTA represents sodium phosphotungstate, and is used at 0.04% by weight. DS means dextran sodium sulfate, u...

Embodiment 2

[0146] (Embodiment 2) Utilize the addition of the present invention Correlation between HDL cholesterol measurement value of serum sample obtained by the first reagent of salt compound and HDL cholesterol measurement value obtained by simple DCM

[0147] Using sodium phosphotungstate as polyanion, containing the present invention The first reagents of salt compounds (reagents A, B, C, D, E, F, G, H, I) were prepared as follows.

[0148] First Reagent (Reagent A)

[0149]

[0150] First Reagent (Reagent B)

[0151]

[0152]

[0153] First Reagent (Reagent C)

[0154]

[0155] First Reagent (Reagent D)

[0156]

[0157] First Reagent (Reagent E)

[0158]

[0159] First Reagent (Reagent F)

[0160]

[0161] First Reagent (Reagent G)

[0162]

[0163] First Reagent (Reagent H)

[0164]

[0165] First Reagent (Reagent I)

[0166]

[0167]

[0168] The HDL cholesterol concentration in the human serum 90 specimen was measured using the HDL ...

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Abstract

The purpose of the present invention is to provide a HDL-C measurement method, whereby it becomes possible to improve the agreement of a measurement value obtained by the method with a measurement value obtained by a reference measurement method such as a DCM even when a sample of a special disease condition such as dyslipidemia is used as a measurement sample. A HDL-C is measured by reacting a measurement sample with a combination of a cholesterol ester hydrolase and a cholesterol oxidase or a combination of a cholesterol ester hydrolase and a cholesterol dehydrogenase in the presence of a compound represented by general formula (I) (wherein R1 represents a linear or branched alkyl group having 8 to 22 carbon atoms or a linear or branched alkenyl group having 8 to 22 carbon atoms; R2, R3 and R4 may be the same as or different from each other and independently represent a linear or branched alkyl group having 1 to 6 carbon atoms or a linear or branched alkenyl group having 2 to 6 carbon atoms; and X- represents an anion) and a polyanion.

Description

technical field [0001] The present invention relates to a method for simply and accurately measuring cholesterol in high-density lipoprotein and a reagent used in the method. Background technique [0002] Cholesterol, neutral fat (hereinafter, sometimes referred to as triglyceride or TG) and phospholipids, which are main lipids in living bodies, exist together with apolipoproteins to form lipoproteins in blood. This lipoprotein is classified into chylomicrons, very low-density lipoprotein (hereinafter, sometimes referred to as VLDL), low-density lipoprotein (hereinafter, sometimes referred to as LDL), and high-density lipoprotein (hereinafter, sometimes referred to as LDL) according to different physical properties. called HDL) and so on. Among them, HDL is known to be a risk prevention factor for various arteriosclerosis represented by coronary arteriosclerosis because it receives cholesterol from various tissues in the living body and is involved in the removal of cholest...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/60C12Q1/26C12Q1/32C12Q1/44G01N33/92
CPCG01N33/92C12Q1/60
Inventor 大田美惠子大野爱子
Owner SEKISUI MEDICAL CO LTD
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