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Adenosine deaminase activity determination method and adenosine deaminase diagnosi kit

A technology of adenosine deaminase and diagnostic kit, which is applied in the field of adenosine deaminase diagnostic kit and determination of adenosine deaminase activity, which can solve problems such as slowness, reduced accuracy of reagent testing, and inconvenient testing

Inactive Publication Date: 2006-05-10
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

[0075] Embodiment one (single agent)

[0076] The adenosine deaminase diagnostic kit of the present embodiment comprises:

[0077] Buffer 80mmol / l

[0078] Adenosine 2mmol / l

[0079] Glutamic acid 5mmol / l

[0080] 2-ketoglutarate 5mmol / l

[0081] Reduced coenzyme 0.2mmo / l

[0082] Adenosine triphosphate 2mmol / l

[0083] Glutamine Synthetase 4000U / l

[0084] Glutamate synthase 4000U / l

[0085] Stabilizer 50% (total volume)

[0086] 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 adenosine deaminase sample to reagent is 1 / 25, reaction direction It is a negative reaction, the delay time is 1 minute, the detection time is 2 minutes, and the theoretical K value is -4180.

[0087] After the sample and reagents are added and allowed to mix, the following reactions occur:

[0088] Adenosine + water adenosine deaminase Inosine + ammonia ion

[0089] AT...

Embodiment 2

[0094] Embodiment two (two doses)

[0095] The adenosine deaminase diagnostic reagent of the present embodiment has:

[0096] Reagent I

[0097] Buffer 100mmol / l

[0098] Glutamic acid 8mmol / l

[0099] 2-ketoglutarate 12mmol / l

[0100] Reduced coenzyme 0.25mmo / l

[0101] Adenosine triphosphate 5mmol / l

[0102] Glutamine Synthetase 7000U / l

[0103] Glutamate synthase 7000U / l

[0104] Stabilizer 50% (total volume)

[0105] Reagent II

[0106] Buffer 100mmol / l

[0107] Adenosine 6mmol / l

[0108] Stabilizer 20mmol / l

[0109] When measuring the activity of adenosine deaminase, the temperature is controlled at 30°C, the reaction time is 15 minutes, the main wavelength of the test is 340nm, and the secondary wavelength of the test is above 405nm. The volume ratio of the tested adenosine deaminase sample to the reagent is 1 / 25. The direction is negative reaction, the delay time is 1 minute, the detection time is 2 minutes, and the theoretical K value is -4180.

[0110] Th...

Embodiment 3

[0117] Embodiment three (three doses)

[0118] The adenosine deaminase diagnostic reagent of the present embodiment is three doses, has:

[0119] Reagent I

[0120] Buffer 120mmol / l

[0121] Glutamic acid 10mmol / l

[0122] 2-ketoglutarate 20mmol / l

[0123] Reduced coenzyme 0.3mmo / l

[0124] Adenosine triphosphate 8mmol / l

[0125] Stabilizer 20mmol / l

[0126] Reagent II

[0127] Buffer 120mmol / l

[0128] Glutamine Synthetase 10000U / l

[0129] Glutamate synthase 10000U / l

[0130] Stabilizer 50% (total volume)

[0131] Reagent III

[0132] Buffer 120mmol / l

[0133] Adenosine 10mmol / l

[0134] Stabilizer 20mmol / l

[0135] Set on the automatic biochemical analyzer: temperature 25°C, reaction time 20 minutes, test main wavelength 340nm, test sub-wavelength above 405nm, the volume ratio of the tested adenosine deaminase sample to the reagent is 1 / 25, and the reaction direction It is a negative reaction, the delay time is 1 minute, the detection time is 2 minutes, and t...

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PUM

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Abstract

This invention relates to a method for measuring the active of the adenosine deamination enzyme and the digonose reagent box for adenosine deamination enzyme, which belongs to the physic checking measuring technique area. This reagent box includes cushion solution, adenosine, glutamic acid, 2-2-ketoglutaric acid,reducing assist enzyme,adenylic triphosphate acid, glutamine synthetic enzyme, glutamic acid synthetic enzyme, stabilizing agent, this method mixes the smple with the reagent by a certain volum proportion to make it fall into the enzyme couplet action, then puts the finally reactant under the biochemistry analyzer to check the absorbenfy changing situation of the main wave to work out the active of the adenosine deamination enzyme. This invention can achive the needed result completely through the biochemistry analyzer, the sensitivity and the definition is high, and it doesn't suffer the pollution from the environment.

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. According to the app...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/25
Inventor 王尔中
Owner 王尔中
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