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Kit for determination of 5'-nucleotidase and preparation method thereof

A nucleotidase and kit technology, applied in the fields of medicine and biochemistry, can solve the problems of complex operation, isotope pollution, and low accuracy

Inactive Publication Date: 2016-11-09
ANHUI IPROCOM BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to provide a test kit for measuring 5'-nucleotidase and its preparation method in order to overcome the defects of complex operation, low accuracy and isotope pollution in the prior art

Method used

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  • Kit for determination of 5'-nucleotidase and preparation method thereof
  • Kit for determination of 5'-nucleotidase and preparation method thereof
  • Kit for determination of 5'-nucleotidase and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] The kit of the present invention includes reagent R1 and reagent R2 two-liquid components independent of each other, wherein

[0063] Reagent R1:

[0064] Good's buffer 80 mmol / L

[0065] 5'-Inosine nucleotide 3.0 KU / L

[0066] Xanthine oxidase 2.1 KU / L

[0067] Purine nucleotide phosphorylase 2.5 KU / L

[0068] Sodium azide 0.4 g / L

[0069] Its solvent is purified water.

[0070] Reagent R2:

[0071] Peroxidase 2.0 KU / L

[0072] 4-Aminoantipyridine 1.9 g / L

[0073] 3-Hydroxy-2,4,6-tribromobenzoic acid 2.8 g / L

[0074] Sodium azide 0.4 g / L

[0075] Its solvent is purified water.

Embodiment 2

[0077] The kit of the present invention includes reagent R1 and reagent R2 two-liquid components independent of each other, wherein

[0078] Reagent R1:

[0079] Good's buffer 20 mmol / L

[0080] 5'-Inosine nucleotide 1.2KU / L

[0081] Xanthine oxidase 3.4 KU / L

[0082] Purine nucleotide phosphorylase 4.5 KU / L

[0083] Sodium azide 0.7 g / L

[0084] Its solvent is purified water.

[0085] Reagent R2:

[0086]Peroxidase 0.4KU / L

[0087] 4-Aminoantipyridine 1.1 g / L

[0088] 3-Hydroxy-2,4,6-tribromobenzoic acid 1.6 g / L

[0089] Sodium azide 0.7 g / L

[0090] Its solvent is purified water.

Embodiment 3

[0092] Kit preparation and method of use

[0093] 1. Prepare the reagent according to the content of the following components:

[0094] Reagent R1:

[0095] Good's buffer 80 mmol / L

[0096] 5'-Inosine nucleotide 3.0 KU / L

[0097] Xanthine oxidase 2.1 KU / L

[0098] Purine nucleotide phosphorylase 2.5 KU / L

[0099] Sodium azide 0.4 g / L

[0100] Its solvent is purified water.

[0101] Reagent R2:

[0102] Peroxidase 2.0 KU / L

[0103] 4-Aminoantipyridine 1.9 g / L

[0104] 3-Hydroxy-2,4,6-tribromobenzoic acid 2.8 g / L

[0105] Sodium azide 0.4 g / L

[0106] Its solvent is purified water.

[0107] 2. Parameter setting of automatic biochemical analyzer

[0108] (a) Detection temperature: 37°C;

[0109] (b) Detection wavelength: main wavelength 505nm, secondary wavelength 700nm;

[0110] (c) Reaction time: 8 minutes, among them, the incubation time is 5 minutes, measure and read the absorbance A1 immediately after adding the reagent R2, read the absorbance A2 after 3 minu...

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Abstract

The invention discloses a kit for determination of 5'-nucleotidase and a preparation method thereof, wherein the kit includes double-liquid components of a reagent R1 and a reagent R2 which are independent and includes the components and the corresponding content: the reagent R1 including a Good's buffer liquid, 5'-hypoxanthine nucleotide, xanthine oxidase, purine nucleotide phosphorylase, sodium azide, and a solvent being purified water; and the reagent R2 including peroxidase, 4-aminoantipyrine, 3-hydroxy-2,4,6-tribromobenzoic acid, sodium azide, and a solvent being purified water. The preparation method includes the steps: according to the component content, preparing the reagents; mixing a to-be-tested sample with the reagent R1 and the reagent R2, and carrying out full reaction; determining the absorbance difference value after the reaction with a fully automatic biochemical analyzer; and calculating the concentration of D-3-hydroxybutyrate in the sample according to the absorbance change value. The kit has the advantages of high accuracy and the like.

Description

technical field [0001] The invention relates to the technical fields of medicine and biochemistry, in particular to a kit for measuring 5'-nucleotidase and a preparation method thereof. Background technique [0002] 5'-nucleotidase (5'-NT), the full name is 5'-ribonucleotide phosphohydrolase. 5'-Nucleotidase is a hydrolase with low substrate specificity and can act on a variety of nucleotides. Mainly involved in the catabolism of nucleic acids, can specifically catalyze nucleoside 5'-nucleotidase and inosine nucleotides, 5'-nucleotidase in the movement of the human body, cell growth and development, fibrin synthesis, improve Epidermal or endothelial shielding function, recirculation and adhesion of neurotransmitting lymphocytes, and immune response all play important roles. [0003] 5'-Nucleotidase is widely distributed in the human body, such as liver, gallbladder, pancreas, intestine, heart, brain, lung, kidney, pituitary, thyroid, prostate, testis and other organs and t...

Claims

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Application Information

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IPC IPC(8): C12Q1/44C12Q1/34C12Q1/28C12Q1/26
CPCC12Q1/44C12Q1/26C12Q1/28C12Q1/34C12Q2326/96
Inventor 蔡晓辉庄庆华吴铮徐运
Owner ANHUI IPROCOM BIOTECH CO LTD
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