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Method for determining activity of adenosine deaminase, and adenosine deaminase detection kit,

The technology of adenosine deaminase and detection kit is applied in the field of adenosine deaminase detection kit to determine the activity of adenosine deaminase, and can solve the problems of difficult operation, high price, increased reagent blank and the like

Inactive Publication Date: 2019-01-01
浙江亚培生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has high sensitivity, wide linear range, good reagent stability, and is widely welcomed by the market. It is a method commonly used at present. However, this method has many reaction steps, and many enzymes and chromogenic reagents are used. As time changes, reagents The blank will increase due to the slow oxidation of the chromogenic reagent, and the cost of the reagent is also high
[0006] In addition, there are many invented methods, application numbers 200410041906.4, 200410041907.9, 200410065300.4, 200410065601.7, etc. The methods adopted have many reaction steps, many internal and external interference factors, and the price of various tool enzymes is very high. Some enzymes are not available on the market and are difficult to operate.

Method used

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  • Method for determining activity of adenosine deaminase, and adenosine deaminase detection kit,
  • Method for determining activity of adenosine deaminase, and adenosine deaminase detection kit,
  • Method for determining activity of adenosine deaminase, and adenosine deaminase detection kit,

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Reagent one:

[0032]

[0033] Reagent 2:

[0034]

[0035] The ratio of reagent 1 to reagent 2 is 2:1, rate method, positive reaction, the wavelength is 340nm, the volume ratio of the tested sample, reagent 1 and reagent 2 is 6: 180: 90, the temperature is 37 ℃, serum and reagent 1 react for 5 minutes , add reagent two, delay time 2 minutes, detect 2 minutes.

[0036] Use this method and xanthine dehydrogenase method two kinds of reagents to detect 50 clinical sera to do contrast test, the experimental result is

[0037] Comparative test of 50 clinical samples

[0038]

[0039] It can be seen that the two methods are highly correlated, r 2 =0.9997

Embodiment 2

[0041] Reagent one:

[0042]

[0043] Reagent 2:

[0044]

[0045] The ratio of reagent one to reagent two is 4:1, rate method, forward reaction, the wavelength is 340nm, the volume ratio of the tested sample, reagent one and reagent two is 6:200:50, the temperature is 37°C, serum and reagent one React for 5 minutes, add reagent 2, delay for 2 minutes, and detect for 2 minutes.

Embodiment 3

[0047] Reagent one:

[0048]

[0049] Reagent 2:

[0050]

[0051] The ratio of reagent 1 to reagent 2 is 3:1, rate method, forward reaction, the wavelength is 340nm, the volume ratio of the tested sample, reagent 1 and reagent 2 is 6: 150: 75, the temperature is 37°C, serum and reagent 1 React for 5 minutes, add reagent 2, delay for 2 minutes, and detect for 2 minutes.

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Abstract

The invention relates to a method for determining the activity of adenosine deaminase, further to an adenosine deaminase detection kit, and belongs to the technical field of medical test determination. According to the method, adenosine used as a substrate is converted and dehydrogenated by using purine nucleoside phosphorylase and xanthine dehydrogenase as tool enzymes, oxidized coenzyme is reduced to form a reduced coenzyme, and the enzyme reaction rate is calculated by detecting the generation amount of the reduced coenzyme I at a wavelength of 340 nm. The method of the present invention has advantages of less reaction steps, less kinds of raw materials, good reagent stability, wide linear range, high sensitivity, good accuracy and convenient promotion.

Description

technical field [0001] The invention relates to a method for measuring the activity of adenosine deaminase, and at the same time, the invention also relates to a detection kit for adenosine deaminase, which belongs to the technical field of medical examination and determination. Background technique [0002] Adenosine deaminase is a kind of sulfhydryl enzyme, which is a key enzyme in the process of purine nucleotide catabolism in cell nucleus metabolism. It catalyzes the oxidation and decomposition of adenine nucleotide, and finally generates uric acid, which is excreted with urine. In humans, adenosine deaminase is mainly derived from lymphocytes and metabolized by the liver. Measuring the level of adenosine deaminase in blood and body fluids is very important for the diagnosis, differential diagnosis and treatment of liver diseases, blood diseases and immune system diseases. As one of the routine examination items of liver function, it can fully reflect the Enzymatic chan...

Claims

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Application Information

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IPC IPC(8): C12Q1/34C12Q1/32C12Q1/48
CPCC12Q1/32C12Q1/34C12Q1/48C12Q2326/96G01N2333/9029G01N2333/91142G01N2333/978
Inventor 李丰盛陈壮永
Owner 浙江亚培生物技术有限公司
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