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Adenosine deaminase diagnostic kit and method for measuring activity concentration of adenosine deaminase

A technology of adenosine deaminase and reagents, which is applied in the field of adenosine deaminase diagnostic kits and the determination of adenosine deaminase activity concentration, and can solve the problems of measurement and difficulty in popularization and application

Inactive Publication Date: 2009-12-23
SUZHOU ANJ BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method cannot be determined by the visible light analyzer in general hospitals, but requires the use of a special ultraviolet light analyzer, so it is difficult to practically popularize and apply

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The adenosine deaminase diagnostic reagent of this embodiment is a single reagent, including:

[0041] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0042] Stabilizer 500mmol / L

[0043] Coenzyme 3mmol / L

[0044] Glutamate oxidase 6000U / L

[0045] Glutamate dehydrogenase 8000U / L

[0046] Adenosine 6mmol / L

[0047] Hydrogen peroxide 5mmol / L

[0048] 2-ketoglutaric acid 5mmol / L

[0049] After the reagents are all dissolved and prepared, they are divided into bottles and freeze-dried to make dry powder reagents; before use, add purified water and reconstitute for use.

[0050] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance ≤ 0.1, test main wavelength 340nm, test subwavelength 405nm, the volume ratio of the tested adenosine deaminase sample to the reagent is 1 / 25, the reaction direction is positive reaction (rising reaction), the delay time is about 1 minute, and the detection time is a...

Embodiment 2

[0053] The adenosine deaminase diagnostic reagent of this embodiment is a double reagent, comprising:

[0054] Reagent 1

[0055] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0056] Stabilizer 50mmol / L

[0057] Coenzyme 3mmol / L

[0058] Adenosine 6mmol / L

[0059] Hydrogen peroxide 5mmol / L

[0060] 2-ketoglutaric acid 5mmol / L

[0061] Reagent 2

[0062] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0063] Stabilizer 500mmol / L

[0064] Glutamate oxidase 6000U / L

[0065] Glutamate dehydrogenase 8000U / L

[0066] After the reagents are all dissolved and prepared, they are divided into bottles to make liquid double reagents, which can be used directly.

[0067] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance ≤ 0.1, test main wavelength 340nm, test subwavelength 405nm, test adenosine deaminase sample and reagent 1, reagent 2 The volume ratio is 2 / 20 / 5, the reaction direct...

Embodiment 3

[0070] The adenosine deaminase diagnostic reagent of this embodiment is three reagents, comprising:

[0071] Reagent 1

[0072] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0073] Stabilizer 50mmol / L

[0074] Coenzyme 3mmol / L

[0075] Adenosine 6mmol / L

[0076] Hydrogen peroxide 5mmol / L

[0077] 2-ketoglutaric acid 5mmol / L

[0078] Reagent 2

[0079] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0080] Stabilizer 500mmol / L

[0081] Glutamate dehydrogenase 8000U / L

[0082] Reagent 3

[0083] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0084] Stabilizer 500mmol / L

[0085] Glutamate oxidase 6000U / L

[0086] After all the reagents are dissolved and prepared, they are divided into bottles to make liquid three reagents, which can be used directly.

[0087] When measuring the activity concentration of adenosine deaminase, set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 m...

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Abstract

The invention relates to an adenosine deaminase diagnostic kit utilizing technologies of an enzymatic recycling method, an enzymatic-colorimetric method and an enzyme-link method, also relates to a method and a principle of measuring activity concentration of adenosine deaminase and compositions and components of reagents, and belongs to the technical field of testing and measuring of medical science. The kit mainly comprises the following compositions: buffer solution, coenzyme, adenosine, hydrogen peroxide, 2-oxoglutaric acid, D-glutamic oxidase, glutamate synthase, and stabilizer; samples are mixed with the reagents in certain volume ratio to perform a series of enzymatic reactions; then reactants are placed under a UV / visible analyzer; and the descending speed of absorbance is tested at the position where dominant wave length is 340nm so as to measure and calculate the activity concentration of the adenosine deaminase.

Description

technical field [0001] The invention relates to a diagnostic kit for adenosine deaminase, and at the same time, the invention also relates to a method for measuring the activity concentration of adenosine deaminase, which belongs to the technical field of inspection 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 applicant knows, the ultra...

Claims

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Application Information

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IPC IPC(8): G01N21/31G01N21/25C12Q1/26C12Q1/68
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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