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Method for measuring 1,5-anhydroglucitol by dehydrogenase

A technology of anhydroglucitol and anhydroglucitol, which is applied in the fields of medicine and biochemistry, and can solve the problems of poor substrate specificity and false increase of measurement results, etc.

Inactive Publication Date: 2014-04-16
WENZHOU PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • 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 dehydrogenase
  • Method for measuring 1,5-anhydroglucitol by dehydrogenase
  • Method for measuring 1,5-anhydroglucitol by dehydrogenase

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Embodiment

[0012] Example: 1,5-anhydroglucitol (1,5-AG) dehydrogenase assay reagent Assay reagent, the buffer solution in component A is particularly preferably pH6.5MES buffer solution, and the buffer solution in component B is particularly preferred pH8.5 Glycine Buffer buffer; the selected surfactant is TritonX-100; the selected preservative is PC300; the selected stabilizer is bovine serum albumin.

[0013] Component A:

[0014]

[0015] Component B:

[0016]

[0017]

[0018] When the reagent of the embodiment is used to determine the sample, the determination method adopted is the two-point endpoint method, the temperature is 37°C, R1: sample: R2 is 240:3:60, the main / secondary wavelength is 560 / 660nm, 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.

[0019] Using this metho...

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Abstract

The invention relates to a method for measuring 1,5-anhydroglucitol by dehydrogenase, which can be widely applied in the field of medical and biochemical technologies. The method comprises the following steps of: converting 1,5-anhydroglucitol (1,5-AG) to be measured and a main interferent, i.e., glucose (Glu), into 1,5-anhydroglucitol-6-phosphate (1,5-AG-6-P) and glucose-6-phosphate (Glu-6-P) by ATP dependent hexohinase (ATP-HK), respectively,and establishing an ATP regeneration system by piruvate kinase (PK) and phosphoenolpyruvate (PEP) in the system so as to ensure that 1,5-AG and Glu is converted into 1,5-AG-6-P and Glu-6-P, respectively. The converted 1,5-A-G-P is specifically dehydrogenated and oxidized by 1,5-anhydroglucitol-6-phosphate dehydrogenase (1,5-AG-6-PDH), and 2-(4-iodophenyl)-5-(2,4-dinitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazole is reduced to a colored substance and then quantified by a full automatic biochemical analyzer by colorimetry.

Description

technical field [0001] The invention relates to a dehydrogenase 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...

Claims

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Application Information

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IPC IPC(8): C12Q1/48C12Q1/32C12Q1/26
Inventor 潘利琴
Owner WENZHOU PEOPLES HOSPITAL
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