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Determination of adenosine deaminase activity and diagnostic kit of adenosine deaminase

A technology of adenosine deaminase and diagnostic kit, which is applied in the field of adenosine deaminase diagnostic kit and the determination of adenosine deaminase activity, can solve the problems of reduced test accuracy of the reagent, unnecessarily urgent, inconvenient test and the like , to achieve the effect of easy operation, simple enzymatic method and high specificity

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

[0078] Embodiment one (single agent)

[0079] The adenosine deaminase diagnostic kit of the present embodiment comprises:

[0080] Buffer 100mmol / l

[0081] Adenosine 1mmol / l

[0082] Monovalent phosphate 2mmol / l

[0083] Nucleoside phosphatase 10000U / l

[0084] Xanthine oxidase 10000U / l

[0085] Ethanol 5mmol / l

[0086] Peroxidase 20000U / l

[0087] Oxidized coenzyme 1mmol / l

[0088] Aldehyde dehydrogenase 10000U / l

[0089] Stabilizer 50% (total volume)

[0090] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, test main wavelength 340nm, test sub-wavelength above 405nm, volume ratio of tested sample to reagent is 1 / 25, reaction direction is positive reaction, delay The time is 1 minute, the detection time is 2 minutes, and the theoretical K value is -4180.

[0091] After the sample and reagents are added and allowed to mix, the following reactions occur:

[0092] Adenosine + water Adenosine deaminase Inosine + ammonia ion

[00...

Embodiment 2

[0099] Embodiment two (two doses)

[0100] The adenosine deaminase diagnostic reagent of the present embodiment has:

[0101] Reagent I

[0102] Buffer 120mmol / l

[0103] Monovalent phosphate 8mmol / l

[0104] Ethanol 8mmol / l

[0105] Oxidized coenzyme 2mmol / l

[0106] Nucleoside phosphatase 20000U / l

[0107] Xanthine oxidase 20000U / l

[0108] Peroxidase 10000U / l

[0109] Aldehyde dehydrogenase 20000U / l

[0110] Stabilizer 50% (total volume)

[0111] Reagent II

[0112] Buffer 120mmol / l

[0113] Adenosine 5mmol / l

[0114] Stabilizer 50mmol / l

[0115] 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 340nm, the secondary wavelength of the test is above 405nm, the volume ratio of the tested sample to the reagent is 1 / 25, and the reaction direction is positive reaction. The delay time is 1 minute, the detection time is 2 minutes, and the theoretical K value is -4180. ...

Embodiment 3

[0124] Embodiment three (three doses)

[0125] The adenosine deaminase diagnostic reagent of the present embodiment is three doses, has:

[0126] Reagent I

[0127] Buffer 80mmol / l

[0128] Monovalent phosphate 5mmol / l

[0129] Ethanol 7mmol / l

[0130] Oxidized coenzyme 2mmol / l

[0131] Stabilizer 30mmol / l

[0132] Reagent II

[0133] Buffer 80mmol / l

[0134] Nucleoside phosphatase 5000U / l

[0135] Xanthine oxidase 5000U / l

[0136] Peroxidase 15000U / l

[0137] Aldehyde dehydrogenase 15000U / l

[0138] Stabilizer 50% (total volume)

[0139] Reagent III

[0140] Buffer 80mmol / l

[0141] Adenosine 3mmol / l

[0142] Stabilizer 20mmol / l

[0143] Set on the automatic biochemical analyzer: temperature 30°C, reaction time 10 minutes, test main wavelength 340nm, test sub-wavelength above 405nm, volume ratio of tested sample to reagent is 1 / 25, reaction direction is positive reaction, delay The time is 1 minute, the detection time is 2 minutes, and the theoretical K value ...

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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, ethanol, peroxidase, oxidized coenzyme, aldehyde dehydrogenase 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 invention relates to a method for measuring the activity of adenosine deaminase, and at the same time, the invention also relates to a diagnostic kit for adenosine deaminase for realizing the method, which belongs to the technical field of medical examination 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 hydrothorax, cerebrospinal fluid, severe combined immunodeficiency disease (SCID), acute and chronic hepatitis, liver 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. As far as the applic...

Claims

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Application Information

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IPC IPC(8): C12Q1/32
Inventor 王尔中
Owner 王尔中
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