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Kit and method for detecting cholesterol concentration in lipoprotein remnant

A technology of detection kits and reagents, applied in the biological field, can solve the problems of laborious, expensive, and not available for use in mainland China, and achieve the effect of simple measurement methods

Inactive Publication Date: 2013-08-07
上海北加生化试剂有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In the last century, Immunoresearch Laboratories of Japan provided RLP-C immunoseparation gel on the market, but the methodological operation is cumbersome, time-consuming, laborious, and expensive, and it is limited to experimental research and is not available for use in mainland China.

Method used

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  • Kit and method for detecting cholesterol concentration in lipoprotein remnant
  • Kit and method for detecting cholesterol concentration in lipoprotein remnant
  • Kit and method for detecting cholesterol concentration in lipoprotein remnant

Examples

Experimental program
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Effect test

Embodiment 1

[0019] The invention provides a detection kit, including two reagents, the first reagent R1 is composed of β-cyclodextrin, anti-human ApoB100 monoclonal antibody, anti-human ApoAI monoclonal antibody, anticyclobate oxidase, 60ul / L cholesterol esterase, 50mmol / L Tris buffer solution, the pH value of described Tris buffer solution is 7.1-7.5; The second reagent R2 is made up of 50mmol / LTris buffer solution, 150mmol / L peroxidase, cholesterol oxidase and 0.06mmol / L 4-aminoantipyrine, the pH value of the Tris buffer is 7.1-7.5.

[0020] Further, the anti-human ApoB100 monoclonal antibody can also be selected from rabbit or goat anti-human ApoB100 polyclonal antibody.

[0021] Further, the ApoAI monoclonal antibody can also be selected from rabbit or goat anti-human ApoAI polyclonal antibody.

Embodiment 2

[0023] Add 2ul of human serum or plasma sample or calibrator to the first reagent R1240ul, incubate at 30°C for 5 minutes,

[0024] Add the second reagent R280ul, incubate at 37°C for 5 minutes, measure the luminosity of the sample (A)

[0025]

[0026] The parameters of the biochemical analyzer determined above are: temperature 37° C., main wavelength 505-550 nm, secondary wavelength 700 nm, and reaction time 10 minutes. The entire operation steps are completed by the biochemical analysis instrument, and the result of the sample can be obtained in only 10 minutes.

[0027] The main parameters detected by RLP-C on Hitachi 7060 analyzer:

[0028] parameters

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Abstract

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.

Description

Technical field: [0001] The invention belongs to the field of biotechnology, and in particular relates to a lipoprotein remnant cholesterol (RLP-C) in serum or plasma, specifically a kit and method for detecting the concentration of lipoprotein remnant cholesterol. Background technique: [0002] Remnant lipoprotein (RLP) is also rich in triglyceride (triglyceride, TG), lipoprotein remnant (TRL) is chylomicron (chylomicron, CM) and very low density lipoprotein (very low density lipoprotein, VLDL ) after lipoprotein lipase (lipoprotein lipase, LPL) hydrolysis gradually lose triglycerides, phospholipids, lipoprotein A (apoprotein A, apoA) and apoC, it is transformed into smaller particles rich in cholesterol, cholesteryl lipid and apoE. [0003] In recent years, with the in-depth understanding of lipoprotein remnants, the clinical significance of RLP determination has been paid more and more attention. A large number of studies have shown that the concentration is significantl...

Claims

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Application Information

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IPC IPC(8): G01N33/50G01N33/577G01N33/92
Inventor 朱晓敏朱慧琳王泉龙杨杰刘颖成刘颖冰
Owner 上海北加生化试剂有限公司
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