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Method and measurement kit for measuring sialic acid by neuraminidase, neuraminic acid zymohexase, and coupled pyruvic oxidase

A technology for detecting kits and sialic acid, which is applied in the measurement of color/spectral characteristics, etc., can solve problems such as interference, low sensitivity, and expensive ketoamine oxidase, and achieve the effects of small variation coefficient, improved sensitivity, and increased stability

Inactive Publication Date: 2019-10-18
浙江曼薇尼生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has high specificity, simple and quick operation, but there are three disadvantages: one is that the sensitivity is not high, because the content of sialic acid is very low, the method has poor reproducibility when detecting low-value serum, and the coefficient of variation is relatively large; Coenzyme Ⅰ is used as a chromogenic reagent, which is easy to oxidize, and the stability of the reagent is poor. Third, it is significantly interfered by pyruvate in the sample.
However, the specificity of ketoamine oxidase is poor, and some amino acids in the sample also react with ketoamine oxidase, and ketoamine oxidase is relatively expensive, which increases the cost of reagents

Method used

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  • Method and measurement kit for measuring sialic acid by neuraminidase, neuraminic acid zymohexase, and coupled pyruvic oxidase
  • Method and measurement kit for measuring sialic acid by neuraminidase, neuraminic acid zymohexase, and coupled pyruvic oxidase
  • Method and measurement kit for measuring sialic acid by neuraminidase, neuraminic acid zymohexase, and coupled pyruvic oxidase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Reagent one:

[0042]

[0043] Reagent 2:

[0044]

[0045] TOOS: N-ethyl-N-(2-hydroxy-3-propanesulfo)-m-toluidine

[0046] Reagent 1: Reagent 2 is 2:1, rate method, positive reaction, wavelength: 546nm, tested sample: Reagent 1: Reagent 2 is 6.0: 180: 90, temperature 37°C, serum reacts with Reagent 1 for 5 minutes, then add reagent Second, delay for 1 minute, and detect the absorbance change rate for 2 minutes.

Embodiment 2

[0048] Reagent one

[0049]

[0050] Reagent 2:

[0051]

[0052] Reagent 1: Reagent 2 is 3:1, rate method, positive reaction, wavelength: 510nm, tested sample: Reagent 1: Reagent 2 is 6.0: 210: 70, temperature 37°C, serum reacts with Reagent 1 for 5 minutes, then add reagent Second, delay for 1 minute, and detect the absorbance change rate for 2 minutes.

Embodiment 3

[0054] Reagent one:

[0055]

[0056]

[0057] Reagent 2:

[0058]

[0059] ADPS: N-ethyl-N-(3-sulfopropyl)-3-methoxyaniline sodium salt.

[0060] Reagent 1: Reagent 2 is 4:1, rate method, positive reaction, wavelength: 546nm, tested sample: Reagent 1: Reagent 2 is 7: 240: 60, temperature 37°C, serum reacts with Reagent 1 for 5 minutes, then add reagent Second, delay for 1 minute, and detect the absorbance change rate for 2 minutes.

[0061] Taking Example 3 as an example, the detection data are as follows:

[0062] The linearity result of the detection kit is

[0063] Theoretical valuemmol / L 0.000 0.625 1.250 1.875 2.500 3.125 3.750 4.375 5.000 Measured valuemmol / L 0.013 0.632 1.256 1.881 2.515 3.129 3.682 4.268 4.706

[0064] Linear equation: y=0.955x+0.066, R 2 = 0.998. See attached figure 1 .

[0065] Detect the purchased quality control product (1.68~2.10~2.52mmol / L), repeat the test 20 times, the results are as follow...

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Abstract

The invention relates to a method for measuring sialic acid by neuraminidase, neuraminic acid zymohexase, and coupled pyruvic oxidase. According to the principle, sialic acid is decomposed by neuraminidase to generate N-acetylneuraminic acid; under the catalysis effect of neuraminic acid zymohexase, the N-acetylneuraminic acid is processed to generate pyruvic acid; and the pyruvic acid is processed under the catalysis effect of pyruvic oxidase to generate hydrogen peroxide; and then Trinder reaction and color displaying are performed. And a kit for measuring the sialic acid can be made.

Description

technical field [0001] The invention relates to a sialic acid detection technology and a sialic acid detection kit, belonging to the field of medical in vitro diagnostic reagents. Background technique [0002] Sialic acid (N-acetylneuraminic acid) is a derivative of 9-carbon monosaccharide, which is mainly distributed in mammals. It was first found in submandibular gland mucin, especially in vascular endothelial cells. The sialic acid in the serum is the total sialic acid, the content is 1.5-2.5mmol / L, including free sialic acid and bound sialic acid, and the bound sialic acid bound to glycoprotein and lipoprotein is called bound sialic acid, which is the main component of serum sialic acid. It is related to cell recognition, differentiation, and adhesion, participates in the composition of receptors, and induces cell proliferation and apoptosis. [0003] Sialic acid is the main component of cell membrane proteins, and has many biological functions, especially in most cases...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/33
CPCG01N21/33
Inventor 李丰盛倪建芬倪忠铨
Owner 浙江曼薇尼生物科技有限公司
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