A reagent for determining the percentage of glycosylated hemoglobin

A technology of glycosylated hemoglobin and hemoglobin, which is applied in the field of reagents for determining the percentage of glycosylated hemoglobin, and can solve problems such as occupation

Active Publication Date: 2016-12-14
AILEX TECH GRP CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the method used to determine the percentage of glycosylated hemoglobin is to measure the percentage of glycosylated hemoglobin in a dual channel, that is, the reaction is carried out in two steps. The first step is to measure the amount of total hemoglobin in the sample, and the second step is to measure the glycated hemoglobin in the sample The amount of hemoglobin, and finally get the ratio of the values ​​after the two-step reaction, that is, the percentage of glycosylated hemoglobin. This form requires two channels for the instrument, and this method brings a lot of inconvenience to the application of the hospital.

Method used

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  • A reagent for determining the percentage of glycosylated hemoglobin
  • A reagent for determining the percentage of glycosylated hemoglobin
  • A reagent for determining the percentage of glycosylated hemoglobin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] After the fresh blood sample was hemolyzed with Triton X-100 hemolysis buffer for 3 minutes, the hemolyzed sample was treated with thermolysin, and the redox reaction was carried out by fructosyl amino acid oxidase, and the generated hydrogen peroxide was developed. Determination.

[0039] The fresh whole blood sample used in the measurement is human whole blood with a glycosylated hemoglobin glycation rate of 10.9%, and the glycosylation rate is obtained by the Tosoh HPLC method.

[0040] Sample pretreatment liquid composition:

[0041]

[0042] Composition of redox reaction solution:

[0043]

[0044] Take 1ml of the above-mentioned 10.9% glycosylated hemoglobin sample after hemolysis, add 4ml of the treatment solution before the sample to it, and react at 37°C for 3 minutes, add 1ml of redox reaction solution to the above reaction solution, and continue at 37°C After reacting for 5 minutes, measure the absorbance at the main wavelength of 700nm and the subwav...

Embodiment 2

[0046] The thermolysin in Example 1 was replaced by Protease E, and the other reaction conditions were the same as in Example 1, and the absorbance was measured on the 10.9% glycosylated hemoglobin sample after hemolysis.

Embodiment 3

[0048] The thermolysin in Example 1 was replaced by subtilisin, and the other reaction conditions were the same as in Example 1, and the absorbance was measured on the 10.9% glycosylated hemoglobin sample after hemolysis.

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Abstract

The invention relates to a pharmaceutical reagent, in particular to a reagent for determining the percentage of glycosylated hemoglobin. The reagent consists of the following substances: a dissolution buffering agent which is used for releasing hemoglobin from red blood cells in a blood sample, protease which can cut off a glycosylated valine or glycosylated polypeptide fragment of an N-terminal end of a beta chain of the glycosylated hemoglobin, an oxidant which can oxidize and eliminate interfering substances in the sample, an oxidant which can denature the hemoglobin and can accelerate the reaction rate of the protease, a reaction accelerator which can accelerate the reaction rate of the protease, fructosyl-amino acid oxidase undergoing oxidation-reduction reaction with the protease which can cut off the glycosylated valine or glycosylated polypeptide fragment of the N-terminal end of the beta chain of the glycosylated hemoglobin in the substance a, a color development agent which can undergo color development reaction with generated hydrogen peroxide, and peroxidase which can catalyze the hydrogen peroxide and the color development agent for the color development reaction. Compared with the prior art, the invention has the advantage that the reagent can directly determine the glycosylated hemoglobin.

Description

technical field [0001] The invention relates to a medical reagent, in particular to a reagent for measuring the percentage of glycosylated hemoglobin. Background technique [0002] Since the glycosylated hemoglobin in the blood reflects the past situation of the blood sugar level in the organism, it is used as an important index for the diagnosis and treatment of diabetes. For the detection of glycosylated hemoglobin, such as: HPLC, electrophoresis, immunoassay, etc. However, for the above-mentioned HPLC method and electrophoresis method, special instruments and equipment are required, and the detection cost is high. At present, it is not popular in my country. For the above-mentioned immune method, although the cost is relatively low, it has the disadvantages of low accuracy and sensitivity. [0003] Therefore, for the determination of glycosylated hemoglobin, the enzymatic method does not require special instruments, the detection cost is low, and the glycosylated hemoglo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/78G01N21/31
Inventor 李子樵孙明礼
Owner AILEX TECH GRP CO LTD
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