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Measuring method of asymmetric dimethylarginine concentration and measuring reagent thereof

A technology of dimethyl arginine and dimethyl arginine dimethylamine, which is applied in the field of medical testing and determination, can solve the problems of automatic analysis that cannot be large-flow, complicated operation, high price, etc., and achieves the detection of large-flow samples , high sensitivity, low cost effect

Inactive Publication Date: 2011-11-16
ZHEJIANG YATAI PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is to overcome the defects that the existing asymmetric dimethylarginine determination method needs special instruments, complex operation, long time-consuming, incapable of automatic analysis of large flow rate, and expensive, and provides a A kind of assay method of asymmetric dimethylarginine concentration, and assay reagent

Method used

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  • Measuring method of asymmetric dimethylarginine concentration and measuring reagent thereof
  • Measuring method of asymmetric dimethylarginine concentration and measuring reagent thereof
  • Measuring method of asymmetric dimethylarginine concentration and measuring reagent thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1 assay reagent

[0041] The assay reagent is in the form of lyophilized powder, and the ingredients are listed in the table below:

[0042] Element

[0043] When in use, dissolve the assay reagent with pH 7, 150mmol / L sodium phosphate buffer, add 100mmol / L mannitol and 0.5g / L bovine serum albumin, and use the buffer to make up to 1 liter. Cooperate with commercially available reagents for the determination of ammonia, and measure the concentration of asymmetric dimethylarginine in the sample to be tested.

Embodiment 2

[0044] The mensuration of embodiment 2 asymmetric dimethylarginine concentration

[0045] Instrument: UV constant temperature spectrophotometer

[0046] Determination reagent: a single reagent in the form of aqueous solution, the ingredients are shown in the following table:

[0047] Element

concentration

HEPES buffer (pH6.4)

20mmol / L

Dimethylarginine dimethylaminohydrolase

2ku / L

Citrulline hydrolase

5ku / L

ornithine carbamoyltransferase

5ku / L

Disodium carbamoyl phosphate

50mmol / L

glutamate dehydrogenase

50ku / L

Thio-NADH

0.28mmol / L

alpha-ketoglutarate

50mmol / L

Mannitol

100mmol / L

bovine serum albumin

0.5g / L

Sodium azide

1g / L

[0048] Samples to be tested: 4 serum samples, the volume ratio of the assay reagent to the sample to be tested is 30:1.

[0049] Assay method: incubate the above assay reagent at 37°C for 5 minutes, place the sample...

Embodiment 3

[0054] The mensuration of embodiment 3 asymmetric dimethylarginine concentration

[0055] Instrument: automatic biochemical analyzer

[0056] Determination Reagent: A two-component kit in the form of aqueous solution, the ingredients are shown in the table below:

[0057]

[0058] Samples to be tested: 6 serum samples, the volume ratio of the assay reagent to the sample to be tested is 30:1.

[0059] Measuring method: add the sample to be tested into reagent 1, incubate at 37°C for 5 minutes, then add reagent 2, and measure after a delay of 2 minutes, measure the absorbance value of the reaction system within 15 minutes, and measure the wavelength at 340nm.

[0060] Measurement and calculation results:

[0061]

[0062] Measurement results of 6 samples:

[0063] Measured object

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Abstract

The invention discloses a measuring method of asymmetric dimethylarginine concentration, comprising the steps of: subjecting a sample to be measured to a reaction in an enzymatic cycling reaction system, measuring and calculating the generation rate of ammonia from the enzymatic cycling reaction, thus obtaining the content of asymmetric dimethylarginine in the sample to be measured. The enzymatic cycling reaction system consists of dimethylarginine dimethylaminohydrolase, citrullinase, ornithine carbamoyltransferase, and carbamyl phosphate or its salt. The invention also discloses a measuring reagent corresponding to the method. In the invention, an enzymology measuring method and a measuring reagent of asymmetric dimethylarginine are established for the first time. Needing no special instrument, the method of the invention has simple operation, high sensitivity and low cost, thus being able to realize rapid and high flow sample detection in clinic. Thus, asymmetric dimethylarginine is probable to become a conventional inspection item in clinic. Therefore, the method provided in the invention is of great scientific and economic value.

Description

technical field [0001] The invention belongs to the technical field of medical examination and measurement, and in particular relates to a method for measuring the concentration of asymmetric dimethylarginine and a reagent for measuring it. Background technique [0002] Asymmetric dimethylarginine (ADMA), a methylated arginine, is an endogenous nitric oxide synthase (Nitric oxide synthase, NOS) inhibitor, which can competitively inhibit the oxidation of Nitrogen (Nitric oxide, NO) synthesis. Asymmetric dimethylarginine is produced by the methylation of intracellular proteins under the action of arginine methyltransferase, followed by hydrolysis. Many cells, such as human endothelial cells, can produce ADMA. A normal human body produces about 300 μmol (about 60 mg) per day, and its concentration in the body is much higher than other endogenous arginine that can also inhibit NOS, so it is considered the most important NOS inhibitor. After ADMA enters the blood circulation f...

Claims

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Application Information

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IPC IPC(8): G01N33/52G01N21/31G01N21/33C12Q1/34C12Q1/48C12Q1/32
Inventor 王学忠
Owner ZHEJIANG YATAI PHARMA
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