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

A technology of adenosine deaminase and reagents, which is applied in the field of adenosine deaminase diagnostic kits and determination of adenosine deaminase activity concentration, and 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

[0039] The adenosine deaminase diagnostic reagent of this embodiment is a single reagent, including:

[0040] Phosphate buffer 100mmol / L

[0041] Stabilizer 500mmol / L

[0042] Coenzyme 3mmol / L

[0043] Adenosine 3mmol / L

[0044] Nucleoside phosphatase 8000U / L

[0045] Xanthine oxidase 8000U / L

[0046] NAD(P)H oxidase 10000U / L

[0047] 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.

[0048] 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, the detection time is about 2 minutes, and the theoretical K value is 4180.

[0049] After adding the sampl...

Embodiment 2

[0051] The adenosine deaminase diagnostic reagent of this embodiment is a double reagent, comprising:

[0052] Reagent 1

[0053] Phosphate buffer 100mmol / L

[0054] Stabilizer 50mmol / L

[0055] Coenzyme 3mmol / L

[0056] Adenosine 3mmol / L

[0057] Reagent 2

[0058] Phosphate buffer 100mmol / L

[0059] Stabilizer 500mmol / L

[0060] Nucleoside phosphatase 8000U / L

[0061] Xanthine oxidase 8000U / L

[0062] NAD(P)H oxidase 10000U / L

[0063] After the reagents are all dissolved and prepared, they are divided into bottles to make liquid double reagents, which can be used directly.

[0064]Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance 0.1, test main wavelength 340nm, test secondary wavelength 405nm, the volume of the tested adenosine deaminase sample and reagent 1, reagent 2 The ratio is 2 / 20 / 5, the reaction direction is positive reaction (rising reaction), the delay time is about 1 minute, the detection time is ab...

Embodiment 3

[0067] The adenosine deaminase diagnostic reagent of this embodiment is three reagents, comprising:

[0068] Reagent 1

[0069] Phosphate buffer 100mmol / L

[0070] Stabilizer 50mmol / L

[0071] Coenzyme 3mmol / L

[0072] Adenosine 3mmol / L

[0073] Reagent 2

[0074] Phosphate buffer 100mmol / L

[0075] Stabilizer 500mmol / L

[0076] Xanthine oxidase 8000U / L

[0077] NAD(P)H oxidase 10000U / L

[0078] Reagent 3

[0079] Phosphate buffer 100mmol / L

[0080] Stabilizer 500mmol / L

[0081] Nucleoside phosphatase 8000U / L

[0082] After all the reagents are dissolved and prepared, they are divided into bottles to make liquid three reagents, which can be used directly.

[0083] When measuring the activity concentration of adenosine deaminase, set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance ≤ 0.1, test main wavelength 340nm, test subwavelength 405nm, measured adenosine deaminase The volume ratio of ammonia enzyme sample t...

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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, coenzyme, adenosine, nucleoside triphosphatase, xanthine oxidase, NAD(P)H oxidase 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 increase 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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