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Double reagent for detecting sialic acid liquid by stable enzyme method and use thereof

A dual-reagent, sialic acid technology, applied in the field of enzymatic determination of sialic acid liquid dual-reagent, can solve the problems of expensive instruments, corrosive reagent components, short stabilization time, etc., to simplify the production process, avoid secondary pollution, bottle small difference effect

Active Publication Date: 2009-07-01
上海微鸿企业管理有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The HPLC method has high sensitivity and good specificity, but the instrument is expensive and is not suitable for clinical batch detection. Although the chemical colorimetric method is cheap, it has low specificity, complicated operation, high temperature extraction, and is not suitable for automatic analysis. The reagent components have Certain corrosiveness and harm to human body
Enzyme analysis method has high specificity, simple and quick operation, accurate and safe, and can be analyzed automatically. The most commonly used enzymatic method is the ultraviolet method coupled with lactate dehydrogenase. Since the automatic analyzer has not been popularized in the past, the reagent consumption is large and the dry powder reagent is complex. The stable time after dissolution is short, the applicable models are few, the user cost is high, and it cannot meet the needs of clinical testing

Method used

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  • Double reagent for detecting sialic acid liquid by stable enzyme method and use thereof
  • Double reagent for detecting sialic acid liquid by stable enzyme method and use thereof
  • Double reagent for detecting sialic acid liquid by stable enzyme method and use thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Reagent 1

[0038] MES buffer pH6.2 100mmol / L

[0039] Neuraminidase 240U / L

[0040] Lactate dehydrogenase 10KU / L

[0041] Surfactant Triton X-100 1ml / L

[0042] Bovine serum albumin 2g / L

[0043] NaN 3 1g / L

[0044] Reagent 2

[0045] Tris buffer pH8.75 50mmol / L

[0046] N-acetylneuraminic acid aldolase 10KU / L

[0047] Reduced coenzyme NADH 0.3mmol / L

[0048] Bovine serum albumin 2g / L

[0049] Glucose dehydrogenase 15U / L

[0050] Glucose 20mmol / L

Embodiment 2

[0052] Reagent 1 includes:

[0053] Neuraminidase 10KU / L

[0054] Lactate dehydrogenase 5KU / L

[0055] BES buffer pH6.0 100mmol / L

[0056] Trehalose 10g / L

[0057] Benzoic acid 1.5g / L

[0058] Reagent 2 includes:

[0059] N-acetylneuraminic acid aldolase 20KU / L

[0060] Reduced coenzyme NADH 0.25mmol / L

[0061]Glucose dehydrogenase 100U / L

[0062] Glucose 100mmol / L

[0063] Glycine buffer pH9.0 75mmol / L

[0064] Trehalose 10g / L

[0065] Methyl hydroxybenzoate 1.5g / L

Embodiment 3

[0067] Reagent 1 includes:

[0068] Neuraminidase 20KU / L

[0069] Lactate dehydrogenase 15KU / L

[0070] Citric acid buffer pH6.0 80mmol / L

[0071] Polyethylene glycol 2g / L

[0072] Gentamicin 3g / L

[0073] Reagent 2 includes:

[0074] N-acetylneuraminic acid aldolase 10KU / L

[0075] Reduced coenzyme NADH 1mmol / L

[0076] Glucose dehydrogenase 20U / L

[0077] Glucose 50mmol / L

[0078] AMP buffer pH8.5 30mmol / L

[0079] Polyethylene glycol 2g / L

[0080] Gentamicin 3g / L

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Abstract

The invention relates to a double reagent, in particular to a liquid double reagent for determining sialic acid by an enzyme method and application thereof. The technical proposal of the liquid double reagent comprises: the steady liquid double reagent for determining the sialic acid by the enzyme method consists of a reagent 1 and a reagent 2, wherein the reagent 1 comprises 0.01 to 50KU / L of neuraminidase, 0.1 to 50KU / L of lactate dehydrogenase, and 25 to 100mmol / L of buffer liquid; and the reagent 2 comprises 0.1 to 50KU / L of N-acetyleuraminic acid aldolase, 0.05 to 5mmol / L of reducing coenzyme NADH, 0.1 to 10000U / L of glucose dehydrogenase, 0.1 to 500mmol / L of glucose, and 5 to 100mmol / L of the buffer liquid. The liquid double reagent has the advantages that the liquid double reagent has convenient use, does not need a dissolving solution to redissolve, has small difference among bottles, ensures that the packaging of the liquid double reagent can adapt to various automatic biochemical analyzers, can avoid secondary pollution, and simplify a production technology method.

Description

technical field [0001] The invention relates to a double reagent, in particular to a liquid double reagent for enzymatically measuring sialic acid and an application thereof. Background technique [0002] Sialic acid, also known as N-acetylneuraminic acid (Sialic Acid, SA), is an important component of cell membrane glycoproteins and exists in two forms combined with cell membrane glycoproteins and glycolipids. Sialic acid is related to many biological functions of organisms. When cell metabolism is abnormal, sialic acid falls off and enters body fluids, resulting in increased sialic acid in the blood. Determination of serum sialic acid concentration can be used as an auxiliary index for cancer diagnosis and curative effect observation index. The dynamic change of SA level in cancer patients is related to the condition, and can be used for early detection of tumor metastasis and recurrence. It is important for the diagnosis and prognosis of liver cirrhosis. Significance, el...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/527
Inventor 夏煜煜
Owner 上海微鸿企业管理有限公司
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