Method for detecting 1,5-anhydro sorbitol and related diagnostic kit

A technology of sorbitan and reagent kits, which is applied in the field of clinical biochemical diagnosis, can solve the problems of high cost of reagent kits, and achieve the effects of ensuring stability, convenient use, and strong removal ability

Active Publication Date: 2011-08-17
BEIJING STRONG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] However, the cost of the kits used in the prior art is high, and further improvement is still needed in terms of stability, sensitivity, and specificity. Therefore, it i

Method used

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  • Method for detecting 1,5-anhydro sorbitol and related diagnostic kit
  • Method for detecting 1,5-anhydro sorbitol and related diagnostic kit
  • Method for detecting 1,5-anhydro sorbitol and related diagnostic kit

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0089] Example one

[0090] Click the ingredients and dosage to configure the 1,5-AG diagnostic kit. Unless otherwise specified, the percentage content in the present invention is a mass-volume ratio.

[0091] Reagent 1

[0092] Phosphate buffer 100mM

[0093] Adenosine diphosphate 5mM

[0094] Magnesium chloride 5Mm

[0095] Adenosine diphosphate-dependent hexokinase 20U / ml

[0096] Tio-NAD 1g / L

[0097] Sodium azide 0.5g / L

[0098] Reagent 2

[0099] Tris buffer 10mM

[0100] 1,5-Anhydroglucitol-6-phosphate dehydrogenase 10U / ml

[0101] NADH 6g / L

[0102] Sodium azide 0.5g / L

[0103] Set on the automatic biochemical analyzer (Olympus-400): reaction temperature 37°C, reaction time 1.5 minutes, test wavelength 405nm, test subwavelength 540nm. The ratio of the measured potassium ion sample to the reagent is: sample: reagent 1: reagent 2 = 5 μl: 180 μl: 60 μl, and the reaction is a lowering reaction.

[0104] First add the sample and reagent 1, the two are automatically mixed in the automatic bio...

Example Embodiment

[0106] Embodiment two:

[0107] According to the first embodiment, the sugar removal system is added to configure a 1,5-AG detection kit.

[0108] Reagent 1

[0109] Tris-HCl 100mmol / L

[0110] Adenosine diphosphate 5mmol / L

[0111] Adenosine triphosphate 5mmol / L

[0112] Hexokinase 5U / ml

[0113] Glucose-6-phosphate 5U / ml

[0114] Hexose phosphate isomerase 10U / ml

[0115] Magnesium chloride 5mmol / L

[0116] Adenosine diphosphate-dependent hexokinase 20U / ml

[0117] Tio-NAD 1g / L

[0118] Sodium azide 0.5g / L

[0119] Reagent 2

[0120] Tris buffer 10mM

[0121] 1,5-Anhydroglucitol-6-phosphate dehydrogenase 10U / ml

[0122] NADH 6g / L

[0123] Sodium azide 0.5g / L

[0124] Use the experimental method of Example 1 to test the sugar removal system.

[0125] Sample

[0126] 1,5-AG pure product solution (150umol / l)

[0127] According to the above data, the sugar-removing system can remove the interference of glucose to the detection, but there is still some interference of glucose to the system.

Example Embodiment

[0128] Example three

[0129] According to the first embodiment, the sugar removal system is added to configure a 1,5-AG detection kit.

[0130] Reagent 1

[0131] Tris-HCl 100mmol / L

[0132] Adenosine diphosphate 5mmol / L

[0133] Adenosine triphosphate 5mmol / L

[0134] Glucokinase 5U / ml

[0135] Glucose-6-phosphate 5U / ml

[0136] Hexose phosphate isomerase 10U / ml

[0137] Magnesium chloride 5mmol / L

[0138] Adenosine diphosphate-dependent hexokinase 20U / ml

[0139] Tio-NAD 1g / L

[0140] Sodium azide 0.5g / L

[0141] Reagent 2

[0142] Tris buffer 10mM

[0143] 1,5-Anhydroglucitol-6-phosphate dehydrogenase 10U / ml

[0144] NADH 6g / L

[0145] Sodium azide 0.5g / L

[0146] Use the experimental method of Example 1 to test the sugar removal system.

[0147] Sample

[0148] According to the above data, the sugar-removing system can remove the interference of glucose on the detection, which is obviously stronger than the ability of removing interference in the third embodiment.

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Abstract

The invention relates to a method for detecting 1,5-anhydro sorbitol and a related diagnostic kit. In the method, a sample and adenosine diphosphate are reacted in the presence of adenosine diphosphate dependent hexokinase serving as a catalyst to form anhydroglucose-6-phosphoric acid and adenylic acid, the anhydroglucose-6-phosphoric acid and thio-oxidative NAD(nicotinamide adenine dinucleotide) are reacted in the presence of 1,5-AG(anhydro glucitol)-6-phosphate dehydrogenase(PDH) serving as a catalyst to form thio-reduced coenzymes and compounds of C6H11O8P, and simultaneously the generated C6H11O8P is reacted in the presence of AG-6-PDH and reduced NAD to form AG-6-P and oxidative coenzymes; and because of the cyclic amplification, the sensitivity of 6-anhydro sorbitol is improved. The invention also provides a detection kit for 1,5-anhydro sorbitol based on the method. The kit is convenient and easy to use, can quickly finish detection on a common ultraviolet/visible light analyzer or semi-/full-automatic biochemical analyzer, does not need special or additional instruments and is low in cost.

Description

technical field [0001] The invention relates to the field of clinical biochemical diagnosis, and relates to a detection method for enzymatic detection of 1,5-sorbitol and a related diagnostic kit. Further, the present invention relates to a method and a kit for detecting 1,5-sorbitol in blood by circulating enzyme method. Background technique [0002] 1,5-anhydroglucitol (1,5-AG), also known as 2-deoxyglucose, 1,5-anhydroglucitol, is a six-carbon monosaccharide with a pyran ring structure. It is the reduced form of glucopyranose (the lack of a hydroxyl group at the C-1 position), forming a C1 chair-shaped oxygen ring at the pyran position, and the molecular formula is C 6 h 12 o 5 (Molecular weight 164.2, CAS number 154-58-5), the structure is very similar to that of glucose, the main difference between the two is that the hydroxyl group at the C-1 position of glucose is replaced by hydrogen, and this substitution is exactly 1,5- Sorbitan is the chemically stable base. ...

Claims

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Application Information

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IPC IPC(8): C12Q1/48C12Q1/32
Inventor 刘瑶杜爱铭刘希蔡华雅
Owner BEIJING STRONG BIOTECH
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