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Adenosine deaminase activity determining method and adenosine deaminase diagnostic reagent

A technology for water adenosine deaminase inosine and adenosine deaminase, which is applied in the field of adenosine deaminase activity determination and adenosine deaminase diagnostic reagents, and achieves high accuracy, easy operation, and easy popularization and application. Effect

Inactive Publication Date: 2005-07-20
SUZHOU ANJ BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its disadvantage is that this system also offsets part of the adenosine deaminase reagent (adenosine deaminase coupling glutamate dehydrogenase reaction must reduce the reduced nicotinamide coenzyme) coenzyme oxidation to oxidized nicotinamide coenzyme) activity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The adenosine deaminase diagnostic reagent of the present embodiment has

[0041] Buffer 100mmol / l

[0042] Adenosine 10mmol / l

[0043] Monovalent phosphate 5mmol / l

[0044] Nucleoside phosphatase 6000U / l

[0045] Xanthine oxidase 8000U / l

[0046] Peroxidase 20000U / l

[0047] 4-amino antiarsenic 2mmol / l

[0048] Carbolic acid 10mmol / l

[0049] When measuring the activity of adenosine deaminase, the temperature is controlled at 36.5-37.5°C, and the ratio of the tested adenosine deaminase sample to the reagent is about 1 / 25. The specific measurement steps are:

[0050] 1. Adenosine + water adenosine deaminase Inosine + ammonia ion

[0051] 2. Inosine + monovalent phosphate nucleoside phosphatase hypoxanthine + ribose 1-phosphate

[0052] 3. Hypoxanthine + water + oxygen xanthine oxidase Urate+Hydrogen Peroxide

[0053] 4. Hydrogen peroxide + 4-amino anti-arsenic + carbolic acid Peroxidase Quinone imine chromogen + water

[0054] 5. Put the final reactant u...

Embodiment 2

[0057] The adenosine deaminase diagnostic reagent of the present embodiment has

[0058] Buffer 100mmol / l

[0059] Adenosine 5mmol / l

[0060] Monovalent phosphate 1mmol / l

[0061] Nucleoside phosphatase 800U / l

[0062] Xanthine oxidase 800U / l

[0063] Peroxidase 800U / l

[0064] 4-amino antiarsenic 0.6mmol / l

[0065] 2,4,6-tribromo-3-hydroxy-benzenesulfonic acid 0.2mmol / l

[0066] When measuring the activity of adenosine deaminase, the temperature is controlled at 29-31°C, and the ratio of the tested adenosine deaminase sample to the reagent is about 1 / 15. The specific measurement steps are:

[0067] 1. Adenosine + water adenosine deaminase Inosine + ammonia ion

[0068] 2. Inosine + monovalent phosphate nucleoside phosphatase hypoxanthine + ribose 1-phosphate

[0069] 3. Hypoxanthine + water + oxygen xanthine oxidase Urate+Hydrogen Peroxide

[0070] 4. Hydrogen peroxide + 4-amino-arsenic + 2,4,6-bromo-3-hydroxy-benzenesulfonic acid Peroxidase Quinone imine chr...

Embodiment 3

[0074] The adenosine deaminase diagnostic reagent of the present embodiment has

[0075] Buffer 100mmol / l

[0076] Adenosine 45mmol / l

[0077] Monovalent phosphate 8mmol / l

[0078] Nucleoside phosphatase 40000U / l

[0079] Xanthine oxidase 42000U / l

[0080] Peroxidase 50000U / l

[0081] 3-methyl-2-benzothiazolone hydrazone 15mmol / l

[0082] Dimethylaniline 18mmol / l

[0083] When measuring the activity of adenosine deaminase, the temperature is controlled at 24-26°C, and the ratio of the tested adenosine deaminase sample to the reagent is about 1 / 200. The specific measurement steps are:

[0084] 1. Adenosine + water adenosine deaminase Inosine + ammonia ion

[0085] 2. Inosine + monovalent phosphate nucleoside phosphatase hypoxanthine + ribose 1-phosphate

[0086] 3. Hypoxanthine + water + oxygen xanthine oxidase Urate+Hydrogen Peroxide

[0087] 4. Hydrogen peroxide + 3-methyl-2-benzothiazolone hydrazone + dimethylaniline perhydrogenase Indamide chromogen + water...

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Abstract

The present invention relates to one kind of adenosine deaminase activity detecting method and the adenosine deaminase diagnosis reagent for the detection, and belongs to the field of medical detection technology. Adenosine, single-valent phosphate, nucleoside phosphorylase, xanthine oxidase, peroxidase and reductive chromoplast are combined to form the main component of the diagnosis reagent. Through serial enzyme linked reactions, the colorless reductive chromoplast is combination oxidized into color dye, so that dye content may be determined with visible light analyzer at 400-600 nm and adenosine deaminase activity may be detected for the diagnosis of several diseases. Compared with available technology, the created dyes have relatively high molar extinction coefficient and thus the present invention has high sensitivity and high precision. In addition, the present invention has no contamination of endogenous matter.

Description

technical field [0001] The invention relates to a method for measuring the activity of adenosine deaminase, and at the same time, the invention also relates to a diagnostic reagent for adenosine deaminase used to realize the method, which belongs to the technical field of medical examination and measurement. Background technique [0002] Medical research shows that adenosine deaminase is a nucleic acid metabolizing enzyme that has an important relationship with the body's cellular immune activity. Therefore, the determination of adenosine deaminase activity is used as an important diagnostic marker for the differential diagnosis of benign and malignant hydrothorax, cerebrospinal fluid, severe combined immunodeficiency disease (SCID), acute and chronic hepatitis, liver cirrhosis, liver cancer and other diseases. [0003] There are many methods for determining the activity of adenosine deaminase, mainly including radionuclide method, physical method and biochemical method. Ac...

Claims

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Application Information

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IPC IPC(8): C12Q1/25C12Q1/42
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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