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Anti-heparin stable alpha-L-fucosidase detection kit and application thereof

A fucosidase and fucosidase technology, which is applied in the measurement of color/spectral properties, etc., can solve the problem of non-resistance to heparin, and achieve the effects of convenient operation, simple preparation and short detection time.

Active Publication Date: 2021-06-18
上海执诚生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The kit provided by the invention effectively removes the interference of heparin, and solves the problem that AFU single reagent is not resistant to heparin

Method used

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  • Anti-heparin stable alpha-L-fucosidase detection kit and application thereof
  • Anti-heparin stable alpha-L-fucosidase detection kit and application thereof
  • Anti-heparin stable alpha-L-fucosidase detection kit and application thereof

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preparation example Construction

[0029] The present invention preferably also provides a preparation method of the above-mentioned kit, comprising: mixing buffer, substrate, surfactant, streptomycin sulfate, stabilizer and preservative, adjusting the pH to 4.5-5.5, and distilling the volume to 1 L, Reagent R is obtained; the reagent R is subpackaged to obtain the above kit. In the present invention, the reagent for adjusting pH preferably includes hydrochloric acid or sodium hydroxide solution. In the present invention, there is no special requirement for the operation of adjusting the pH, and operations well known to those skilled in the art can be used.

[0030] The present invention also provides the application of the above kit in detecting α-L-fucosidase. In the present invention, the detected samples preferably include samples containing heparin. The detection of α-L-fucosidase by the kit of the present invention is for non-diagnostic purposes, and only for obtaining an intermediate value of α-L-fucos...

Embodiment 1

[0035] An anti-heparin stable α-L-fucosidase detection kit, the preparation of the kit: 300mmol acetic acid-sodium acetate buffer solution, 1g M-G-2-chloro-p-nitrophenol-α-L- Fucoside, 30g of Triton 100, 0.03g of streptomycin sulfate, 0.1g of L-cysteine ​​and 2g of Proclin 300 were sequentially added into the beaker and stirred until completely dissolved. After adjusting the pH to 5.2 with hydrochloric acid and sodium hydroxide solution, Dilute to 1L to obtain reagent R, and divide reagent R into 60mL / bottles to obtain kits.

Embodiment 2

[0037] An anti-heparin stable α-L-fucosidase detection kit, the preparation of the kit: 300mmol acetic acid-sodium acetate buffer solution, 1g M-G-2-chloro-p-nitrophenol-α-L- Add fucoside, 30g Triton 100, 0.05g streptomycin sulfate, 0.05g L-cysteine ​​and 2g Proclin 300 into the beaker and stir until completely dissolved, adjust the pH to 5.2 with hydrochloric acid and sodium hydroxide solution, Dilute to 1L to obtain reagent R, and divide reagent R into 60mL / bottles to obtain kits.

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Abstract

The invention relates to the technical field of detection reagents, in particular to an anti-heparin stable alpha-L-fucosidase detection kit and application thereof. The kit comprises 0.03-0.1 g / L of streptomycin sulfate, 50-500 mmol / L of a buffer solution, 0.1-3 g / L of a substrate, 1-50 g / L of a surfactant, 0.001-0.1 g / L of a stabilizer and 0.3-3 g / L of a preservative; and the substrate is prepared from 2-chloro-p-nitrophenol-alpha-L-fucoside or M-G-2-chloro-p-nitrophenol-alpha-L-fucoside. Streptomycin sulfate with proper concentration is added into the alpha-L-fucosidase detection reagent, so that interference of heparin in a sample is effectively removed, and the problem that an AFU single reagent is not resistant to heparin is solved.

Description

technical field [0001] The invention relates to the technical field of detection reagents, in particular to a heparin-resistant and stable α-L-fucosidase detection kit and its application. Background technique [0002] α-L-fucosidase (AFU) is a lysosomal acid hydrolase, which is widely distributed in various tissues, cells and body fluids in the human body, especially in tissues such as liver and kidney. AFU has good sensitivity and high positive rate in the diagnosis of hepatocellular carcinoma, which is more than three times the positive rate of AFP. It has great diagnostic value for AFP-negative cases and small cell liver cancer, and is a useful indicator for the diagnosis of early primary liver cancer. [0003] At present, the methods for detecting AFU mainly include fluorescence method, endpoint colorimetric method, and rate method. Fluorescence method is to use fucosidase compound to release 4-methylumbelliferone that can emit fluorescence after hydrolysis by fucosida...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31
CPCG01N21/31
Inventor 王青泉朱荫华张璇申文君蔡顺斐扈金舟张秀丽李宝玉
Owner 上海执诚生物科技有限公司
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