Preparation method for calibration matter for calibrating apolipoprotein A1 and apolipoprotein B

An apolipoprotein and calibrator technology, which is applied in the field of biomedicine, can solve the problems of general stability, long time consumption, easy turbidity of the solution, etc., and achieves the effect of improving biological safety and simple method.

Active Publication Date: 2014-09-03
玉环市南方试剂有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] When preparing Apoa1 and Apob calibrants by traditional methods, human serum is directly subjected to ultracentrifugation, which not only requires many times of centrifugation and takes a long time,

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment one: 1. Collect 1.5L of people's mixed serum, [after detecting hepatitis B virus surface antigen (HBsAg), hepatitis C virus surface antibody (HCV antibody) and human immunodeficiency virus type I and type II antibody (HIV antibody ) were negative], of which 0.5L was used as matrix serum, and the other 1L was used to extract LDL and HDL components.

[0035]2. Extraction of LDL: Add 4ml of 10% polyanion solution to 1L of mixed human serum, mix well, immediately form LDL-polyanion polymer, divide into 4 tubes, centrifuge at 4000 rpm for 20 minutes, (the upper layer is collected in In another container for HDL extraction), pour the centrifuge tube on the filter paper for 15 minutes, drain the remaining supernatant, add 10ml of 0.5 mol / L potassium oxalate solution to each centrifuge tube, stir with a glass rod, Polyanions are precipitated, centrifuged at 4000 rpm for 20 minutes, the supernatant is a yellow and transparent LDL component, the concentration is about ...

Embodiment 2

[0039] Embodiment two: 1. collect people's mixed serum 1.5L, [after detecting hepatitis B virus surface antigen (HBsAg), hepatitis C virus surface antibody (HCV antibody) and human immunodeficiency virus type Ⅰ and type Ⅱ antibody (HIV antibody ) were negative], of which 0.5L was used as matrix serum, and the other 1L was used to extract LDL and HDL components.

[0040] 2. Extraction of LDL: add 4ml of 6% polyanion solution to 1L of mixed human serum, mix well, immediately form LDL-polyanion polymer, divide into 4 tubes, centrifuge at 4000 rpm for 20 minutes, (the upper layer is collected in In another container for HDL extraction), pour the centrifuge tube on the filter paper for 15 minutes, drain the remaining supernatant, add 10ml of 0.5 mol / L potassium oxalate solution to each centrifuge tube, stir with a glass rod, Polyanions are precipitated, centrifuged at 4000 rpm for 20 minutes, the supernatant is a yellow and transparent LDL component, the concentration is about 20 t...

Embodiment 3

[0044] Embodiment three: 1. collect people's mixed serum 1.5L, [after detecting hepatitis B virus surface antigen (HBsAg), hepatitis C virus surface antibody (HCV antibody) and human immunodeficiency virus type Ⅰ and type Ⅱ antibody (HIV antibody ) were negative], of which 0.5L was used as matrix serum, and the other 1L was used to extract LDL and HDL components.

[0045] 2. Extraction of LDL: Add 4ml of 15% polyanion solution to 1L of mixed human serum, mix well, immediately form LDL-polyanion polymer, divide into 4 tubes, centrifuge at 4000 rpm for 20 minutes, (the upper layer is collected in In another container for HDL extraction), pour the centrifuge tube on the filter paper for 15 minutes, drain the remaining supernatant, add 10ml of 0.5 mol / L potassium oxalate solution to each centrifuge tube, stir with a glass rod, Polyanions are precipitated, centrifuged at 4000 rpm for 20 minutes, the supernatant is a yellow and transparent LDL component, the concentration is about 2...

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Abstract

The invention discloses a preparation method for a calibration matter for calibrating apolipoprotein A1 and apolipoprotein B. The essentials of the technical scheme are as follows: the preparation method comprises the following steps: (1) adding a multi-anionic compound and divalent metal ions into blood serum to carry out pre-treatment; (2) extracting low density lipoprotein (LDL); (3) extracting high density lipoprotein (HDL); (4) mixing protein and base blood serum to prepare the calibration matter; (5) defining the value of the calibration matter. The preparation method has the advantages that the extraction of LDL and HDL components in the blood serum by a multi-anionic compound and divalent metal ion sub-step precipitation method is simplified; the content of the ApoB and the content of the ApoA1 are detected; the calibration matter can be mixed into the blood serum of healthy people according to the needed amount so that the ApoA1 and the ApoB reach the predicated content; the method is simple; a stabilizer and a turbidity-preventing agent are added into the mixed blood serum so that the prepared freeze-dried calibration matter is clarified after being re-dissolved; the stability is good and a base material effect does not exist; the calibration matter is diluted into five different concentrations and can be used for non-linear calibration of determining the ApoA1 and the ApoB on an automatic analyzer.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a method for preparing apolipoprotein A1 and apolipoprotein B calibrator. Background technique [0002] Clinically, the determination of serum apolipoprotein A1 (ApoA1) and apolipoprotein B (ApoB) is often used as the risk estimation of cardiovascular and cerebrovascular diseases, the monitoring of the efficacy of drug treatment, and the diagnosis of certain abnormal lipoproteinemia. The commonly used serum ApoA1 and ApoB assay kits in China all adopt Immunoturbidimetric assay (ITA), because it is fast, accurate, precise and suitable for various types of automatic biochemical analyzers, especially suitable for clinical laboratories. Determination of large batches of specimens. However, multiple points (generally 5 different concentrations) are required for nonlinear calibration during the determination, so a calibrator with a high-end concentration of the ApoA1 and ApoB measurement ra...

Claims

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Application Information

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IPC IPC(8): G01N33/96
CPCG01N33/92G01N33/96
Inventor 杨昌国
Owner 玉环市南方试剂有限公司
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