Method, substrate and reagents for alpha-N-acetylglucosaminidase activity detection

A technology of acetylglucosamine and acetylglucosamine, which is applied in a field and can solve the problems of inability to accurately diagnose patients' diseases, complex symptoms, and inability to truly analyze causative factors.

Inactive Publication Date: 2014-01-01
北京中科非凡生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The difficulty in diagnosing mucopolysaccharidosis type IIIB is reflected in many aspects: first, because the disease is a multi-system disease, involving genetics, nerves, muscles, blood, bones, eyes and other organs or systems, the symptoms are complex, and are related to many common diseases. There are similar clinical manifestations, so it is difficult for doctors to accumulate diagnostic experience from symptom manifestations, so missed diagnosis, misdiagnosis and delayed diagnosis are common problems
Secondly, the existing routine examinations and other auxiliary methods in the hospital are difficult to achieve the purpose of diagnosis, such as bone marrow aspiration, physical examination, electrocardiogram,

Method used

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  • Method, substrate and reagents for alpha-N-acetylglucosaminidase activity detection
  • Method, substrate and reagents for alpha-N-acetylglucosaminidase activity detection
  • Method, substrate and reagents for alpha-N-acetylglucosaminidase activity detection

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Experimental program
Comparison scheme
Effect test

Embodiment

[0063] The α-N-acetylglucosaminidase detection reagent of this embodiment is a single reagent, including

[0064] Disodium hydrogen phosphate / citric acid buffer (pH4.5) 100mmol / L

[0065] Stabilizer 0.1%

[0066] R-α-N-acetylglucosamine 0.1mmol / L

[0067] The substrate in the reagent is R-α-N-acetylglucosamine, and R is 4-methylumbelliferone.

[0068] The sample of α-N-acetylglucosaminidase to be tested is human white blood cells, the total protein dosage is 0.05ug / ul, the volume ratio of protein to reagent is 1 / 9, the sample and reagent are added and mixed to make it react, and the reaction time After 24 hours, stop solution was added to terminate the reaction after the reaction, and the fluorescence value at excitation wavelength 360nm (Ex=360nm) and emission wavelength 460nm (Em=460nm) was detected with fluorescence detection equipment. The average fluorescence reading value of the reagent blank is 30, and the average fluorescence reading value of normal human leukocyte ...

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Abstract

The present invention provides a method for alpha-N-acetylglucosaminidase (EC3.2.1.50) activity detection, and further provides a substrate and reagents for alpha-N-acetylglucosaminidase activity determination. The method, the substrate and the reagents can be used for analyzing and determining alpha-N-acetylglucosaminidase activity in a sample (including human body fluid or tissue or cell samples) requiring determination, and are mainly used in the field of clinical laboratory. The determination reagent prepared by using the method has characteristics of convenience, rapidness and high sensitivity, and is easily promoted and applied.

Description

technical field [0001] The invention belongs to the technical field of biomedicine and at the same time belongs to the field of clinical diagnosis and detection of genetic metabolic diseases. Specifically, the present invention provides a method for measuring the enzyme activity of α-N-acetylglucosaminidase (α-N-acetylglucosaminidase, EC 3.2.1.50), and the present invention also provides a method as α-N-acetylglucosaminidase A substrate for the detection of glucosaminidase (α-N-acetylglucosaminidase, EC 3.2. 1.50) Reagents for enzymatic activity. [0002] Background technique [0003] Lysosome is an organelle wrapped by a unit membrane in human cells. It contains a variety of acid hydrolytic enzymes. More than 60 kinds have been found so far, such as protease, nuclease, glycosidase, lipase, phosphatase, lysozyme and other enzymes. In the cell, lysosomal enzymes control the digestion of various endogenous and exogenous macromolecular substances, such as nucleic acids, pro...

Claims

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Application Information

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IPC IPC(8): C12Q1/34C07H17/075G01N21/64
Inventor 孙宏博
Owner 北京中科非凡生物技术有限公司
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