5'-ribonucleotide hydrolytic enzyme detection kit and preparation method thereof

A ribonucleotide and detection kit technology, applied in the field of biomedicine, can solve the problems of unsuitable routine inspection, reaction susceptible to exogenous interference, instability and the like

Active Publication Date: 2012-10-24
乐普(北京)诊断技术股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] There are many methods for the determination of 5’-ribonucleotide hydrolase, but most of them are unstable, and the reaction is susceptible to external interference, so it is not suitable for routine testing

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The kit of the present invention can be an example of a double reagent, wherein:

[0026] Reagent 1:

[0027] Glycine buffer 80mmol / L

[0028] Color developer 10mmol / L

[0029] Purine nucleoside phosphorylase 0.5KU / L

[0030] Xanthine oxidase 0.8KU / L

[0031] Peroxidase 0.6KU / L

[0032] Reagent 2:

[0033] 5'-Inosine nucleotide 30mmol / L

[0034] 4-aminoantipyrine solution 10mmol / L

[0035] in:

[0036] (1) Glycine buffer is 35mM glycine, 0.05% Tween-20, 10mM disodium edetate, 0.05% thimerosal, 0.4M NaCl, pH6.5-10;

[0037] (2) The chromogenic reagent is a phosphate buffer containing 20mM 3-methyl-N,N-dipropanesulfonate sodium aniline;

[0038] (3) The 4-aminoantipyrine solution is a chloroform solution containing 25 mM 4-aminoantipyrine.

Embodiment 2

[0039] Embodiment 2: Kit use method

[0040] 1. Reagent preparation, the reagent can be a liquid double reagent, ready to use after opening the bottle, of which:

[0041] Reagent 1:

[0042] Glycine buffer 80mmol / L

[0043] Color developer 10mmol / L

[0044] Purine nucleoside phosphorylase 0.5KU / L

[0045] Xanthine oxidase 0.8KU / L

[0046] Peroxidase 0.6KU / L

[0047] Reagent 2:

[0048] 5'-Inosine nucleotide 30mmol / L

[0049] 4-aminoantipyrine solution 10mmol / L

[0050] 2. Automatic biochemical analyzer parameter setting

[0051] (a) Detection temperature: 37°C

[0052] (b) Detection wavelength: main wavelength is 550nm; secondary wavelength is 700nm;

[0053] (c) Reaction time: 10 minutes, wherein the incubation time is 5 minutes, the reaction time is 3 minutes, and the detection time is 2 minutes.

[0054] 3. Detection steps (all completed in the automatic biochemical analyzer)

[0055] (a) Mix 225 μL reagent 1 and 6 μL serum sample.

[0056] (b) Incubate the mix...

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Abstract

The invention discloses a 5'-ribonucleotide hydrolytic enzyme detection kit. A kit body comprises a glycine buffer solution, a color developing agent prepared from 3- (m-tolidino) dipropane sulfonic acid disodium salt, purine nucleoside phosphorylase, xanthine oxidase, peroxidase, 5'-hypoxanthine nucleotide and 4-aminoantipyrine solution. A sample is mixed with a reagent according to a certain volume ratio for a series of reaction, reactants are placed under a semi- / full-automatic biochemical analyzer, and the change speed of light absorptivity in the position of a main wavelength of 550nm is detected so as to measure and calculate the concentration of the 5'-ribonucleotide hydrolytic enzyme. The detection kit has the advantages of accuracy, stability and convenience.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, in particular to a detection kit for detecting human 5'-ribonucleotide hydrolase. Background technique [0002] 5'-ribonucleotide hydrolase is an enzyme that plays a key role in the catabolism of 5'-inosine nucleotides. It can catalyze the degradation of 5'-inosine nucleotides into inosine nucleoside, and then undergo Nucleoside phosphorylase catalyzes hypoxanthine, which is eventually oxidized to the metabolite uric acid. [0003] 5'-ribonucleotide hydrolase is a special hydrolase that can specifically hydrolyze 5'-nucleotides linked to 5'-phosphates of pentoses to generate nucleosides and phosphates. Widely present in the liver and various tissues, its increased activity is mainly seen in diseases of the hepatobiliary system, such as obstructive jaundice, primary and secondary liver cancer, etc. [0004] Serum 5'-ribonucleotide hydrolase activity in normal adults has no difference between...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/44C12Q1/34C12Q1/28C12Q1/26
Inventor 张雷
Owner 乐普(北京)诊断技术股份有限公司
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