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Adenosine deaminase diagnosis reagent kit and method for determining adenosine deaminase activity concentration

A technology of adenosine deaminase and adenosine, applied in the field of determination of adenosine deaminase activity concentration, adenosine deaminase diagnostic kit, can solve problems such as difficult promotion, application and measurement

Inactive Publication Date: 2008-12-17
沈丽华
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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

[0043] The adenosine deaminase diagnostic reagent of this embodiment is a single reagent, comprising:

[0044] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0045] Stabilizer 500mmol / L

[0046] Reduced coenzyme 0.25mmol / L

[0047] Adenosine 5mmol / L

[0048] Pyruvate 15mmol / L

[0049] Indole 10mmol / L

[0050] Acetyl-CoA 20mmol / L

[0051] Tryptophanase 10000U / L

[0052] Dopa decarboxylase 10000U / L

[0053] Pyruvate dehydrogenase 12000U / L

[0054] 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.

[0055] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance 1.8±0.7, test main wavelength 340nm, test secondary wavelength 405nm, the volume ratio of the tested adenosine deaminase sample to the reagent is 1 / 25, the reaction direction is negative reaction (falling reacti...

Embodiment 2

[0058] The adenosine deaminase diagnostic reagent of this embodiment is a double reagent, comprising:

[0059] Reagent 1

[0060] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0061] Stabilizer 50mmol / L

[0062] Reduced coenzyme 0.25mmol / L

[0063] Adenosine 5mmol / L

[0064]Pyruvate 15mmol / L

[0065] Indole 10mmol / L

[0066] Acetyl-CoA 20mmol / L

[0067] Reagent 2

[0068] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0069] Stabilizer 500mmol / L

[0070] Tryptophanase 10000U / L

[0071] Dopa decarboxylase 10000U / L

[0072] Pyruvate dehydrogenase 12000U / L

[0073] After the reagents are all dissolved and prepared, they are divided into bottles to make liquid double reagents, which can be used directly.

[0074] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance 1.8±0.7, test main wavelength 340nm, test secondary wavelength 405nm, tested adenosine deaminase sample and r...

Embodiment 3

[0077] The adenosine deaminase diagnostic reagent of this embodiment is three reagents, comprising:

[0078] Reagent 1

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

[0080] Stabilizer 50mmol / L

[0081] Reduced coenzyme 0.25mmol / L

[0082] Adenosine 5mmol / L

[0083] Pyruvate 15mmol / L

[0084] Indole 10mmol / L

[0085] Acetyl-CoA 20mmol / L

[0086] Reagent 2

[0087] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0088] Stabilizer 500mmol / L

[0089] Dopa decarboxylase 10000U / L

[0090] Pyruvate dehydrogenase 12000U / L

[0091] Reagent 3

[0092] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0093] Stabilizer 500mmol / L

[0094] Tryptophanase 10000U / L

[0095] After all the reagents are dissolved and prepared, they are divided into bottles to make liquid three reagents, which can be used directly.

[0096] When measuring the activity concentration of adenosine deaminase, set on the automatic bi...

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Abstract

The invention relates to a kit for diagnosing adenosine deaminase by utilizing the technologies of the enzymic colorimetric method and the enzyme linked immunosorbent assay (ELISA). The invention further relates to a method, a principle and the composition and the components of a reagent for mensurating the active concentration of the adenosine deaminase, and belongs to the technology field of medical inspection and measurement. The main components of the kit include a buffer solution, reduced coenzyme, adenosine, pyruvic acid, indole, acetyl coenzyme A, tryptophanase, dopa decarboxylase, pyruvate dehydrogenase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet / visible light analyzer, and the velocity of the decrease in absorbance at 340nm of the dominant wavelength is detected, thereby mensurating the active 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 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] The methods for determining the activity of adenosine deaminase mainly include radionuclide method, physical method and biochemical method. As far as the applicant knows, th...

Claims

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Application Information

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IPC IPC(8): G01N33/50G01N21/31C12Q1/26C12Q1/00
Inventor 王尔中
Owner 沈丽华
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