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Adenosine deaminase diagnosing reagent case and method for detecting adenosine deaminase active density

The technology of adenosine deaminase and diagnostic kit is applied in the field of adenosine deaminase diagnostic kit and the determination of adenosine deaminase activity concentration, which can solve the problem of reducing the accuracy of reagent testing, inconvenient testing and slowing down. urgent problem

Inactive Publication Date: 2008-01-02
SUZHOU ANJ BIOTECHNOLOGY CO LTD
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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

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

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

[0045] Stabilizer 50mmol / L

[0046] Reduced coenzyme 0.25mmol / L

[0047] Glutamate dehydrogenase 50000U / L

[0048] Glutamate oxidase 10000U / L

[0049] α-ketoglutarate 16mmol / L

[0050] 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.

[0051] 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 sample to be tested is 1 / 25, the reaction The direction is negative reaction (falling reaction), the detection method is the rate method, the delay time is about 1 minute, and the detection time is about 2 minutes.

[0052] Afte...

Embodiment 2

[0054] The adenosine deaminase diagnostic reagent of this embodiment is a double reagent, comprising:

[0055] Reagent 1

[0056]Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0057] Stabilizer 50mmol / L

[0058] Reduced coenzyme 0.25mmol / L

[0059] α-ketoglutarate 12mmol / L

[0060] Reagent 2

[0061] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0062] Stabilizer 50mmol / L

[0063] Glutamate dehydrogenase 40000U / L

[0064] Glutamate oxidase 15000U / L

[0065] After the reagents are all dissolved and prepared, they are divided into bottles to make liquid double reagents, which can be used directly.

[0066] 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 sample to reagent 1 and reagent 2 is 2 / 20 / 5, the reaction direction is negative reaction (decreasing reaction), the d...

Embodiment 3

[0069] The adenosine deaminase diagnostic reagent of this embodiment is three reagents, comprising:

[0070] Reagent 1

[0071] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0072] Stabilizer 50mmol / L

[0073] Reduced coenzyme 0.25mmol / L

[0074] α-ketoglutarate 10mmol / L

[0075] Reagent 2

[0076] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0077] Stabilizer 50mmol / L

[0078] Glutamate dehydrogenase 60000U / L

[0079] Reagent 3

[0080] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0081] Stabilizer 50mmol / L

[0082] Glutamate oxidase 20000U / L

[0083] After all the reagents are dissolved and prepared, they are divided into bottles to make liquid three reagents, which can be used directly.

[0084] When measuring the activity concentration of adenosine deaminase, set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance 1.8±0.7, test main wavelength...

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Abstract

The invention discloses an adenosine deaminase diagnosing agent box through enzyme circulating augmentation method, density agent composition and component in the medical detecting and testing technical domain, which comprises the following parts: buffer, adenosine, alpha-ketoglutaric acid, glutamate dehydrogenase, glutamic oxidase, reduction type coenzyme and stabilizer, wherein the samples are blended according to certain bulk rate to do series of enzymatic reaction and places the reactants under ultraviolet / visible analyzer, which detects the absorbance dropping speed of major wavelength at 340nm to calculate the activity density of the adenosine deaminase. The invention improves the sensitivity and precision, which is convenient to spread and apply.

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 determining the activity concentration of adenosine deaminase, 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 applicant's ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/34
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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