A method for detecting alpha-L-fucosidase vitality and agent therefor

A fucosidase and fucose technology, applied in the field of biochemistry, can solve the problems of affecting the detection effect, long detection time, negative detection results, etc., and achieve the effects of simple and convenient operation method, short reaction time and high sensitivity

Inactive Publication Date: 2006-02-08
王贤理
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This technology allows us to quickly determine if there are substances that may affect biological processes accurately with only 40 nm wavelength lights. It works well even when exposed to sunlight or other sources like UV rays. Its technical effect is improved efficiency over existing methods while also being easier and more user friendly than current technologies such as fluorescence analysis.

Problems solved by technology

This patented technical problem addressed by this patent relates to improving current techniques such as immunochromatography assays for measuring alpha - L-fuzymer lyases, including their ability to accurately measure low levels of these compounds during disease states like hepatitis B virus carriers' circulation system syndrome and metastasis.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1 (detection method):

[0029] Addition

blank tube

Assay tube

Reagent 1 (μl)

200

200

Distilled water (μl)

25

——

Sample (μl)

——

25

Mix well and incubate at 30°C for 9 minutes

Reagent 2 (μl)

50

50

Mix well, extend for 60 seconds at 30°C, measure the absorbance change in the linear phase of the reaction at a wavelength of 340nm, and then

Then calculate the average absorbance change rate ΔA / min per minute.

[0030] Calculation formula:

[0031] TV: total reaction volume SV: sample volume

[0032] ε: Millimolar extinction coefficient of NADH at 340nm: 6.22

[0033] P: the light diameter of the cuvette

Embodiment 2

[0034] Embodiment 2 (detection method):

[0035] Basic operation of single reagent assay:

[0036] Basic operation of single reagent assay:

[0037] Addition

[0038] The calculation formula is the same as before.

Embodiment 3

[0039] Embodiment 3 (two reagents):

[0040] Reagent A (R1)

[0041] Buffer MES 20mmol / L

[0042] Preservative thiourea 2.5mmol / L

[0043] Stabilizer dimercaptothreitol 0.005mmol / L

[0044] Stabilizer EDTA.2Na 0.005mmol / L

[0045] Surfactant NP-40 3ml / L

[0046] Substrate a-L-(-)-fucose-D-glucoside 5mmol / L

[0047] Reagent B (R2)

[0048] Buffer Tris 80mmol / L

[0049] Preservative sodium azide 2.5mmol / L

[0050] Stabilizer Trehalose 10mmol / L

[0051] Stabilizer EDTA.2Na 2mmol / L

[0052] Coenzyme NADP 8mmol / L

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PUM

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Abstract

The invention relates to a method for measuring the slurryª‡-L-fucose glycosidase (AFU) vitality and its agent in the field of biochemistry technology. The invention proportionally adds the tested sample into the agent and mixes them at 30-40 degree about 1.5-10 minutes, then it uses full automation biochemistry analysis to measure the linear changing rate of the absorbance at 340nm to compute the vitality of the sample AFU. The agent for measuring the slurryª‡-L-fucose glycosidase (AFU) vitality comprises a buffer liquid, a preservation, a stabilizer, a bottom object, a surface activator and oxid-type niacinamide cobamaide analogy.

Description

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Claims

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Application Information

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Owner 王贤理
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