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Determination of adenosine deaminase activity and diagnostic kit of adenosine deaminase

A technology for adenosine deaminase and diagnostic kits, which is applied in the field of adenosine deaminase diagnostic kits and the determination of adenosine deaminase activity, and can solve the problems of inconvenient testing, slowness, and reduced accuracy of reagent testing, etc. , to achieve the effects of easy operation, improved sensitivity, and easy promotion and application

Inactive Publication Date: 2006-03-15
王尔中
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

One of its disadvantages is that this system also offsets part of the adenosine deaminase reagent (adenosine deaminase coupled glutamate dehydrogenase reaction must Reduced nicotinamide coenzyme is oxidized to oxidized nicotinamide coenzyme), which greatly reduces the accuracy of the reagent test
The second disadvantage is that the reagent must be reacted for a period of time before the formal test in order to produce enough reduced nicotinamide coenzyme, which results in slow results and brings great inconvenience to the test

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] Example one (single dose)

[0097]The adenosine deaminase diagnostic kit of this embodiment includes:

[0098] Buffer 80mmol / l

[0099] Adenosine 1mmol / l

[0100] Monovalent phosphate 2mmol / l

[0101] Nucleoside Phosphatase 5000U / l

[0102] Xanthine Oxidase 5000U / l

[0103] Peroxidase 5000U / l

[0104] Urease 5000U / l

[0105] 4-amino anti-arsenic 2mmol / l

[0106] Carbonic acid 10mmol / l

[0107] Stabilizer 30% (total volume)

[0108] Set on the automatic biochemical analyzer: temperature 37℃, reaction time 10 minutes, test main wavelength 495~505nm, test sub-wavelength above 600nm, volume ratio of test sample to reagent is 1 / 25, reaction direction is positive reaction , The delay time is 1 minute, and the detection time is 2 minutes.

[0109] After adding the sample and reagents and mixing them, the following reactions occur:

[0110] Adenosine + water Adenosine Deaminase Inosine + Ammonia

[0111] Inosine + Monovalent Phosphate Nucleoside Phosphatase Hypoxanthine+1-phosp...

Embodiment 2

[0117] Example two (double dose)

[0118] The adenosine deaminase diagnostic reagents of this embodiment include:

[0119] Reagent I

[0120] Buffer 100mmol / l

[0121] Monovalent phosphate 3mmol / l

[0122] Nucleoside Phosphatase 20000U / l

[0123] Xanthine Oxidase 20000U / l

[0124] Peroxidase 20000U / l

[0125] Urease 20000U / l

[0126] Stabilizer 40% (total volume)

[0127] 4-amino anti-arsenic 1mmol / l

[0128] Reagent II

[0129] Buffer 100mmol / l

[0130] Adenosine 3mmol / l

[0131] Stabilizer 30mmol / l

[0132] 2,4,6-Sabromo-3-hydroxy-benzenesulfonic acid 1mmol / l

[0133] When measuring adenosine deaminase activity, the temperature is controlled at 30°C, the reaction time is 10 minutes, the main wavelength of the test is 546nm, and the subwavelength of the test is above 630nm. The volume ratio of the test sample to the reagent is 1 / 25, and the reaction direction is positive reaction. The delay time is 1 minute, and the detection time is 2 minutes.

[0134] The specific measurement s...

Embodiment 3

[0142] Example three (three doses)

[0143] The diagnostic reagent for adenosine deaminase in this embodiment consists of three doses, including:

[0144] Reagent I

[0145] Buffer 120mmol / l

[0146] Monovalent phosphate 5mmol / l

[0147] Stabilizer 25mmol / l

[0148] 3-methyl-2-benzothiazolone hydrazone 2mmol / l

[0149] Reagent II

[0150] Buffer 120mmol / l

[0151] Nucleoside Phosphatase 50000U / l

[0152] Xanthine Oxidase 50000U / l

[0153] Peroxidase 50000U / l

[0154] Urease 50000U / l

[0155] Stabilizer 50% (total volume)

[0156] Reagent III

[0157] Buffer 120mmol / l

[0158] Adenosine 5mmol / l

[0159] Dimethylaniline 2mmol / l

[0160] Stabilizer 30mmol / l

[0161] Set on the automatic biochemical analyzer: temperature 25℃, reaction time 10 minutes, test main wavelength 578nm, test sub-wavelength above 700nm, volume ratio of test sample to reagent is 1 / 25, reaction direction is positive reaction, delay The time is 1 minute, and the detection time is 2 minutes.

[0162] The specific ...

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Abstract

The invention is to determine the activity of the Adenosine deaminase and the reagent box of it. The box includes buffer, adenosine, phosphate, nucleoside phosphorylase, xanthine oxidase, peroxidase, urease, reduced chromogen and stabilizer. According to the proper ratio to mix the reagents, so the reagent can react and we can get the activity of the enzyme by detecting the absorbance of the main wave.

Description

Technical field [0001] The present invention relates to a method for measuring the activity of adenosine deaminase. At the same time, the present invention also relates to a diagnostic kit for adenosine deaminase for realizing the method, which belongs to the technical field of medical testing and determination. 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 pleural fluid, cerebrospinal fluid, severe combined immunodeficiency (SCID), acute and chronic hepatitis, cirrhosis, liver cancer and other diseases. [0003] The methods for determining the activity of adenosine deaminase mainly include radionuclide method, physical method and biochemical method. According to the applicant’s u...

Claims

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Application Information

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IPC IPC(8): C12Q1/42
Inventor 王尔中
Owner 王尔中
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