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Adenosin-deaminase diagnostic kit and adenosine deaminease activity determining method

A technology of adenosine deaminase and diagnostic kit, which is applied in the field of determination of adenosine deaminase activity and adenosine deaminase diagnostic kit, which can solve the problems of reduced test accuracy, inconvenient test, slow time, etc. , to achieve the effects of shortening test time, high test accuracy, and easy promotion and application

Inactive Publication Date: 2005-07-20
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

[0048] The adenosine deaminase diagnostic kit of the present embodiment comprises:

[0049] Reagent I

[0050] Phosphate buffer 100mmol / l

[0051] 2-ketoglutarate 10mmol / l

[0052] Reduced coenzyme 0.2mmo / l

[0053] Glutamate dehydrogenase 200000U / l

[0054] Stabilizer 20mmol / l

[0055] Reagent II

[0056] Phosphate buffer 100mmol / l

[0057] Adenosine 2mmol / l

[0058] Set on the automatic biochemical analyzer 1: temperature 37°C, reaction time 10 minutes, initial absorbance less than 1.8, test main wavelength 340mn, test secondary wavelength 450mn, volume ratio of adenosine deaminase sample to double reagent It is 1 / 25, the reaction direction is negative reaction, the delay time is 1 minute, the detection time is 2 minutes, and the theoretical K value is -4180.

[0059] After the sample and reagents are added and allowed to mix, the following reactions occur:

[0060] Adenosine + water adenosine deaminase Inosine + ammonia ion

[0061] Ammonium + 2-oxoglutarate + redu...

Embodiment 2

[0064] The adenosine deaminase diagnostic reagent of the present embodiment has

[0065] Reagent I

[0066] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 180mmol / l

[0067] 2-ketoglutarate 40mmol / l

[0068] Reduced coenzyme 0.25mmo / l

[0069] Glutamate dehydrogenase 400000U / l

[0070] Stabilizer 20mmol / l

[0071] Reagent II

[0072] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 180mmol / l

[0073] Adenosine 40mmol / l

[0074] When measuring the activity of adenosine deaminase, the temperature is controlled at 37°C, the reaction time is 10 minutes, the initial absorbance is less than 1.7, the main wavelength of the test is 340mn, and the secondary wavelength of the test is 450mn. The ratio of the tested adenosine deaminase sample to the reagent is about 4 / 50, the reaction direction is negative reaction, the delay time is 1 minute, the detection time is 2 minutes, and the theoretical K value is -2170.

[0075] The specific measurement steps are:

[0...

Embodiment 3

[0081] The adenosine deaminase diagnostic reagent of the present embodiment is single reagent, has

[0082] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 50mmol / l

[0083] 2-ketoglutarate 5mmol / l

[0084] Reduced coenzyme 0.25mmo / l

[0085] Glutamate dehydrogenase 10000U / l

[0086] Adenosine 3mmol / l

[0087] Stabilizer 20mmol / l

[0088] When measuring the activity of adenosine deaminase, the temperature is controlled at 37°C, the reaction time is 10 minutes, the initial absorbance is less than 1.7, the main wavelength of the test is 340mn, and the secondary wavelength of the test is 450mn. The ratio of the tested adenosine deaminase sample to the reagent is about 1 / 12.5, the reaction direction is negative reaction, the delay time is 5 minutes, the detection time is 2 minutes, and the theoretical K value is -2170.

[0089] The specific measurement steps are:

[0090] 1. Adenosine + water adenosine deaminase Inosine + ammonia ion

[0091] 2. Ammonium ion + 2-o...

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Abstract

The present invention relates to one kind of diagnostic adenosine deaminase reagent kit and method of measuring activity of adenosine deaminase, and belongs to the field of medical detection technology. The reagent kit of the present invention includes buffering liquid, 2-ketoglutarate, reduced coenzyme, glutamate dehydrogenase and adenosine. The sample is mixed with the reagent in certain volume ratio to produce reduced enzyme-linked reaction, and the reaction product is set under visible light analyzer to detect the absorbance descending speed or main wavelength or subsidiary wavelength, so as to obtain the activity of adenosine deaminase via calculation. The present invention obtains the determination results in visible light analyzer, and has high sensitivity, high precision and no contamination of endogenous matter.

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 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, the ultraviolet ...

Claims

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Application Information

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IPC IPC(8): C12Q1/25
Inventor 王尔中
Owner 王尔中
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