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Method for measuring 1,5-anhydroglucitol by oxidase

A technology of anhydroglucitol and its determination method, which is applied in the direction of biochemical equipment and methods, and the determination/inspection of microorganisms, and can solve problems such as falsely increased determination results and poor substrate specificity

Inactive Publication Date: 2014-04-16
WENZHOU PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

Due to the poor substrate specificity of the PROD enzyme, the 6-phosphate glucose generated in the reaction is easily reconverted into glucose when the content of pyruvate kinase is insufficient, and the generated glucose is oxidized by the PROD enzyme to participate in Trindre's reaction and develop color, thereby causing 1,5-AG assay results are falsely elevated

Method used

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  • Method for measuring 1,5-anhydroglucitol by oxidase
  • Method for measuring 1,5-anhydroglucitol by oxidase
  • Method for measuring 1,5-anhydroglucitol by oxidase

Examples

Experimental program
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Embodiment

[0012] Embodiment: the dehydrogenase assay reagent of 1,5-anhydroglucitol (1,5-AG) Assay reagent, particularly preferred damping fluid is pH8.0 Tris-HCl damping fluid; Especially selected preferred surfactant is TritonX-100; a particularly preferred preservative is PC300; a preferred stabilizer is bovine serum albumin.

[0013] Component A:

[0014]

[0015] Component B:

[0016]

[0017] When the reagent of the embodiment is used to measure the sample, the measuring method adopted is the two-point endpoint method, the temperature is 37 °C, R1: sample: R2 is 240: 5: 80, the main / minor wavelength is 600 / 700nm, and R1 is added After the sample or standard is incubated at the measuring temperature for 180-300 seconds, read the absorbance at the first point, then add R2 and continue to incubate for 300 seconds, then read the absorbance at the second point.

[0018] Using this method and the literature method (Chen Guojun, et al. Chinese Journal of Medical Laboratory, 1999,...

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Abstract

The invention relates to a method for measuring 1,5-anhydroglucitol by oxidase, which can be widely applied in the field of medical and biochemical technologies. The method comprises the following steps of: converting glucose (Glu) in a sample into fructose-6-phosphate (Fru-6-P) by glucokinase (GK) and phosphoglucose isomerase (PGI), and establishing an ATP regeneration system by pyruvate kanise (PK) and phosphoenolpyruvate (PEP) in the system so as to ensure that Glu is converted into Fru-6-P. In the sample, 1,5-AG reacts in the presence of pyranose oxidase (PROD) to produce 1,5-fructosan (1,5-AF) and H2O2, and then H2O2 is colored and quantified via a Trinder's system..

Description

technical field [0001] The invention relates to an oxidation method for measuring 1,5-anhydroglucitol (1,5-AG), which can be widely used in the technical fields of medicine and biochemistry. Background technique [0002] The concentration of 1,5-anhydroglucitol in serum is not affected by immediate diet and glucose intake, and can quickly and sensitively reflect changes in total blood sugar within 3-5 days. Serum 1,5-AG decreased significantly in diabetic patients. When diabetes is well controlled, the daily recovery rate of serum 1,5-AG is very stable, regardless of treatment methods, gender, age, weight, disease time and serum 1 , the impact of the initial concentration of 5-AG, the range of recovery and the sensitivity and specificity to the diagnosis of diabetes are significantly greater than the commonly used diabetes diagnostic indicators glycosylated hemoglobin (HbAlc) and fructosamine (GSP). [0003] According to literature reports, methods for determining 1,5-AG in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/533C12Q1/48C12Q1/26
Inventor 潘利琴
Owner WENZHOU PEOPLES HOSPITAL
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