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Method for detecting glycosylated hemoglobin by chemiluminescence

A technology of glycosylated hemoglobin and chemiluminescence, which is applied in the field of detection of glycosylated hemoglobin by chemiluminescence technology, which can solve the problems of cumbersome operation steps, low reproducibility, and susceptibility to environmental influences

Active Publication Date: 2017-09-22
QINGDAO UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods still have many shortcomings, such as expensive instruments, cumbersome operation steps, low reproducibility and susceptibility to environmental influence, etc.

Method used

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  • Method for detecting glycosylated hemoglobin by chemiluminescence
  • Method for detecting glycosylated hemoglobin by chemiluminescence
  • Method for detecting glycosylated hemoglobin by chemiluminescence

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0029] Example 1: A method for detecting glycosylated hemoglobin by chemiluminescence technology

[0030] 1. Experimental part

[0031] 1.1 Instruments and reagents

[0032] 1.1.1 Instruments and equipment

[0033] DHG blast drying oven (Shanzhi Instrument Equipment Co., Ltd., Shanghai); AR224CN Ohaus analytical balance (Qingdao Zhonghe Hengxin Electronics Co., Ltd., Qingdao); THZ constant temperature oscillation box (Jiayuan Xingye Technology Co., Ltd., Beijing) ; RFL-1 ultra-weak chemiluminescence detector (Ruimai Analytical Instrument Co., Ltd., Xi'an); Anke-TGL-16C Feiweng brand high-speed centrifuge (Anting Scientific Instrument Factory, Shanghai).

[0034] 1.1.2 Reagents

[0035] Glycated hemoglobin was purchased from Sigma-Aldrich Co. LLC. 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS), chloroauric acid (HAuCl 4 ) was purchased from Sigma; the carboxyl magnetic beads with a particle size of 0.5 μm and a concentration o...

example 2

[0048] Example 2: Sample Analysis

[0049] The sample solution containing glycosylated hemoglobin is tested according to the method in step 1.5, and the content of glycosylated hemoglobin can be obtained according to the chemiluminescence intensity and the standard curve obtained in step 1.5.

[0050] According to the method of the invention, the glycated hemoglobin content was measured, and the method was evaluated by the standard addition method. The recovery rate of the sample was 97.0-103.2%. The measurement results are shown in Table 1. The method of the present invention has precision High degree of characteristics.

[0051] Table 1. Sample analysis results

[0052]

[0053] a 7 measurements

[0054] b Unit: mg / mL

example 3

[0055] Example 3: Method Sensitivity Comparison

[0056] Using LumAuNPs-H 2 o 2 For the detection system, when the other steps are the same, the method of the present invention is used to detect the glycated hemoglobin, and the detection limit of the determination is 0.8 ng / mL. It shows that a method for detecting glycosylated hemoglobin by chemiluminescence technology proposed by the present invention has high sensitivity. When the concentration of glycosylated hemoglobin was 1.0ng / mL, LumAuNPs-hydroxylamine-O-sulfonic acid was used as the chemiluminescence detection system for chemiluminescence measurement, and the chemiluminescence intensity produced was LumAuNPs-H 2 o 2 4 times that of the chemiluminescent detection system ( image 3 ).

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Abstract

The invention belongs to the field of analytical chemistry and particularly relates to a method for detecting glycosylated hemoglobin by chemiluminescence. The method adopts the principle that chloroauric acid is reduced by luminol to obtain luminol colloid gold nanoparticles LumAuNPs, and LumAuNPs are modified with aptamer complementary sequences DNA to obtain chemiluminescent probes; aptamers DNA are fixed by taking magnetic beads as carriers to obtain DNA modified magnetic beads; the chemiluminescent probes and the DNA modified magnetic beads are incubated to obtain chemiluminescence sensors; when a target object (glycosylated hemoglobin) exists, the aptamers DNA and glycosylated hemoglobin act with each other, and the chemiluminescent probes drop off the surfaces of the magnetic beads; and after magnetic separation, hydroxylamine-O-sulfonic acid is added into a separating medium, and LumAuNPs-hydroxylamine-O-sulfonic acid is taken as a chemiluminescent system, chemiluminescence assay is performed, and the target glycosylated hemoglobin is detected according to the generated chemiluminiscence. The method has the advantages of being simple and being high in sensitivity.

Description

technical field [0001] The invention belongs to the field of medical examination, and in particular relates to a method for detecting glycated hemoglobin by chemiluminescence technology. Background technique [0002] Hemoglobin is a protein (Hb, HGB) responsible for carrying oxygen in higher organisms. Hemoglobin consists of four chains, two alpha chains and two beta chains, each chain has a ring of heme containing an iron atom. Oxygen binds to iron atoms and is transported by the blood. The characteristics of hemoglobin are: in places with high oxygen content, it is easy to combine with oxygen; in places with low oxygen content, it is easy to separate from oxygen. This characteristic of hemoglobin enables red blood cells to transport oxygen. Glycosylated hemoglobin (HbA1c) is the product of the combination of hemoglobin and blood sugar in red blood cells in human blood. The combination of blood sugar and hemoglobin to form glycosylated hemoglobin is an irreversible reac...

Claims

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Application Information

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IPC IPC(8): G01N33/72G01N33/543G01N21/76
CPCG01N21/763G01N33/54326G01N33/723
Inventor 混旭刘冰茹王冠中张慧
Owner QINGDAO UNIV OF SCI & TECH
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