Assay kit and assay method for adenosine deaminase

The technology of adenosine deaminase and kit is applied in the field of adenosine deaminase assay kit and its assay, which can solve the problems of large amount of stabilizer, complicated reagents, detection interference, etc., and achieve enhanced enzyme activity stability, The effect of avoiding precipitation and improving stability

Active Publication Date: 2018-10-23
广州市伊川生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are many ADA activity assay kits, and the reagent components and assay effects are not the same, but the stability of purine nucleoside phosphorylase and xanthine oxidase is generally poor, and it is easy to cause assay results due to enzyme inactivation during storage. Unreliable
Chinese patent application CN 101514358 discloses a method for improving the stability of adenosine deaminase reagent by coupled enzymatic reaction, including adding calcium edetate, iron edetate and sodium molybdate to the basic reagents , ammonium molybdate, sodium glutamate, bovine serum albumin or superoxide dismutase as a stabilizer step can improve the stability of the reagent, but the large amount of stabilizer may affect the rapid progress of the reaction and even cause detect interference
[0004] Chinese patent application CN 102586397 discloses a kit for enzymatic detection of adenosine deaminase, which consists of reagent 1 and reagent 2, reagent 1 includes buffer, toos, Triton x-100, sodium benzoate, FeCl 3 , PNP, Xot, Peo, glyceraldehyde, sodium arsenate; Reagent 2 includes disodium hydrogen phosphate, KCl, mannitol, proclin300, adenosine, 4-aminotipyrine; Accelerate adenosine by adding accelerators to reagent 1 For the reaction of glycoside deaminase, adding mannitol to reagent 2 improves the stability of the reagent, but the reagent is more complicated and the stability is still not ideal

Method used

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  • Assay kit and assay method for adenosine deaminase
  • Assay kit and assay method for adenosine deaminase
  • Assay kit and assay method for adenosine deaminase

Examples

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Effect test

Embodiment 1

[0029] Example 1 Adenosine deaminase assay kit (formula 2)

[0030] Adenosine deaminase assay kit includes reagent R1 and reagent R2, said reagent R1 includes the following components and their concentrations: TRIS 12g / L, 6N HCL 20g / L, potassium sorbate 1g / L, xanthine oxidase 3KU / L, purine nucleoside phosphorylase 3KU / L, 4-aminoantipyrine 60mg / L, peroxidase 6KU / L, sodium aspartate 50g / L; the reagent R2 includes the following components and Its concentration: TRIS 12g / L, 6N HCL 20g / L, potassium sorbate 1g / L, adenosine 5g / L, N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3-methyl Aniline sodium salt 0.5g / L, glycerin 200ml / L.

Embodiment 2

[0031] Embodiment two adenosine deaminase assay kit

[0032] Adenosine deaminase assay kit includes reagent R1 and reagent R2, said reagent R1 includes the following components and their concentrations: TRIS 15g / L, 6N HCL 18g / L, potassium sorbate 1g / L, xanthine oxidase 2KU / L, purine nucleoside phosphorylase 2KU / L, 4-aminoantipyrine 55mg / L, peroxidase 3KU / L, sodium aspartate 40g / L; The reagent R2 includes the following components and Its concentration: TRIS 15g / L, 6N HCL 18g / L, potassium sorbate 1g / L, adenosine 3g / L, N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3-methyl Aniline sodium salt 1g / L, glycerin 300ml / L.

Embodiment 3

[0033] Example three adenosine deaminase assay kit

[0034] Adenosine deaminase assay kit includes reagent R1 and reagent R2, said reagent R1 includes the following components and their concentrations: TRIS 10g / L, 6N HCL 22g / L, potassium sorbate 1g / L, xanthine oxidase 4KU / L, purine nucleoside phosphorylase 4KU / L, 4-aminoantipyrine 65mg / L, peroxidase 8KU / L, sodium aspartate 60g / L; the reagent R2 includes the following components and Its concentration: TRIS 10g / L, 6N HCL 22g / L, potassium sorbate 1g / L, adenosine 6g / L, N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3-methyl Aniline sodium salt 2g / L, glycerin 200ml / L.

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Abstract

The invention provides an assay kit for adenosine deaminase. The assay kit comprises a reagent R1 and a reagent R2. The reagent R1 comprises TRIS, HCL, xanthine oxidase, purine nucleoside phosphorylase, 4-aminoantipyrine, peroxidase and sodium aspartate; and the reagent R2 comprises TRIS, HCL, adenosine, N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3-methylaniline sodium slat and glycerol. The assay kit ofthe invention belongs to the technical field of biological detection, is simple in reagent composition, good in stability, rapid in reaction and low in cost, and can accurately detect adenosine deaminase; and the detection results of the assay kit have no obvious difference from the detection results of listed products.

Description

technical field [0001] The invention belongs to the technical field of biological detection, in particular to an adenosine deaminase assay kit and a assay method thereof. Background technique [0002] Adenosine deaminase (ADA) is an important enzyme in the purine nucleotide cycle, which can catalyze the conversion of adenosine into inosine, and then generate hypoxanthine through the action of nucleoside phosphorylase, and its metabolism The final product is uric acid, which plays an important role in nucleic acid metabolism. ADA is widely distributed in the small intestinal mucosa, spleen, liver, kidney, and other tissues and cells of the human body. ADA in serum is mainly derived from the liver and belongs to the cytoplasmic enzyme of liver cells. When hepatocytes are damaged, necrotic or have increased membrane permeability, they can lead to increased serum ADA activity. Therefore, ADA can be used as a good biochemical index for auxiliary diagnosis of liver cirrhosis an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/48C12Q1/34C12Q1/28C12Q1/26
CPCC12Q1/26C12Q1/28C12Q1/34C12Q1/48C12Q2326/96G01N2333/978
Inventor 刘光华杨玉军陈楚华刘秋明
Owner 广州市伊川生物科技有限公司
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