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Adenosine deaminase (ADA) detection reagent kit and preparation method thereof

A technology of adenosine deaminase and detection kit, which is applied in the field of detection kits for detecting human adenosine deaminase, and can solve problems such as serum ammonia-containing interference

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

This method is also useless due to the interference of serum ammonia and non-specific oxidation caused by excessive NADPH in the test system.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The kit of the present invention can be an example of a double reagent, wherein:

[0028] Reagent 1:

[0029] Glycine buffer 80mmol / L

[0030] Color developer 10mmol / L

[0031] Purine nucleoside phosphorylase 0.5KU / L

[0032] Xanthine oxidase 0.8KU / L

[0033] Peroxidase 0.6KU / L

[0034] Reagent 2:

[0035] Adenosine 30mmol / L

[0036] 4-aminoantipyrine solution 10mmol / L

[0037] in:

[0038] (1) Glycine buffer is 35mM glycine, 0.05% Tween-20, 10mM disodium edetate, 0.05% thimerosal, 0.4M NaCl, pH6.5-10;

[0039] (2) The chromogenic reagent is a phosphate buffer solution containing 20 mM 3-methyl-N, N-dipropanesulfonate sodium aniline;

[0040] (3) The 4-aminoantipyrine solution is a chloroform solution containing 25 mM 4-aminoantipyrine.

Embodiment 2

[0041] Embodiment 2: Kit use method

[0042] 1. Reagent preparation, the reagent can be a liquid double reagent, ready to use after opening the bottle, of which:

[0043] Reagent 1:

[0044] Glycine buffer 80mmol / L

[0045] Color developer 10mmol / L

[0046] Purine nucleoside phosphorylase 0.5KU / L

[0047] Xanthine oxidase 0.8KU / L

[0048] Peroxidase 0.6KU / L

[0049] Reagent 2:

[0050] Adenosine 30mmol / L

[0051] 4-aminoantipyrine solution 10mmol / L

[0052] 2. Automatic biochemical analyzer parameter setting

[0053] (a) Detection temperature: 37°C

[0054] (b) Detection wavelength: main wavelength is 550nm; secondary wavelength is 700nm;

[0055] (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.

[0056] 3. Detection steps (all completed in the automatic biochemical analyzer)

[0057] (a) Mix 180 μL reagent 1 and 5 μL serum sample.

[0058] (b) Incubate the mixed solution ...

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Abstract

The invention discloses a detection reagent kit for detecting human ADA. A reagent kit body contains a glycine buffer solution, a color-developing agent prepared with 3-methyl-N,N-dipropyl sodium sulfonate aniline, purine nucleoside phosphorylase, xanthine oxidase, peroxidase, adenosine and a 4-aminoantipyrine solution. A sample and a reagent are mixed in a certain volume proportion and subjected to a series of reactions, then reactants are placed under a semi-automatic / full-automatic biochemical analyzer, and the change rate of the absorbency at a position of 550nm dominant wavelength is detected, so that the concentration of the ADA is measured and calculated. The reagent kit has the advantages of being accurate, stable and convenient.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, in particular to a detection kit for detecting human adenosine deaminase. Background technique [0002] Adenosine deaminase (ADA) is an enzyme that plays a key role in the catabolic process of purine nucleoside. It can catalyze the degradation of adenine nucleoside into inosine nucleoside, which is then catalyzed by nucleoside phosphorylase into hypoxanthine, and finally Oxidized to the metabolite uric acid. There are three isozymes of adenosine deaminase, namely adenosine deaminase 1, adenosine deaminase 1+cp and adenosine deaminase 2. [0003] The distribution of adenosine deaminase in animals is highest in the cecum, small intestinal mucosa and spleen, and is also found in fibroblasts, amniocytes, liver, kidney, lung, skeletal muscle, etc. ADA in blood mainly exists in blood cells. For example, the activity of red blood cells, lymphocytes and granulocytes is about 40 to 70 times that of ...

Claims

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Application Information

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