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Detection method for activity of ornithine carbamoyltransferase, and detection reagent thereof

An ornithine carbamoyltransferase and a determination method are applied in the field of medical inspection and determination, and can solve the problems of automatic analysis that cannot be large-flow, high price, complicated operation, etc., and achieve large-flow sample detection, low cost, and high sensitivity. high effect

Inactive Publication Date: 2011-11-02
王学忠
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to overcome the defects that the existing ornithine carbamoyltransferase assay method requires special instruments, complicated operation, time-consuming, incapable of large-flow automatic analysis, and expensive, and provides a A method for assaying the activity of ornithine carbamoyltransferase, and assay reagents

Method used

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  • Detection method for activity of ornithine carbamoyltransferase, and detection reagent thereof
  • Detection method for activity of ornithine carbamoyltransferase, and detection reagent thereof
  • Detection method for activity of ornithine carbamoyltransferase, and detection reagent thereof

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Experimental program
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Effect test

Embodiment 1

[0038] Embodiment 1 assay reagent

[0039] The assay reagent is in the form of lyophilized powder, and the ingredients are listed in the table below:

[0040] Element

[0041] When used, dissolve the assay reagent with pH 7, 150 mmol / L sodium phosphate buffer. Cooperate with commercially available reagents for the determination of ammonia, and measure the activity of ornithine carbamoyltransferase on the samples to be tested.

Embodiment 2

[0042] The determination of embodiment 2 ornithine carbamoyltransferase activity

[0043] Instrument: UV constant temperature spectrophotometer

[0044] Determination reagent: a single reagent in the form of aqueous solution, the ingredients are shown in the following table:

[0045] Element

[0046] During the measurement, there is no fixed requirement for the ratio of the reagent to the sample, but it should be consistent within the same measurement batch. Here, the reagent:sample ratio is 3000:100. Add the sample to be tested to the reagent pre-warmed at 37°C, and measure after a delay of 2 minutes. Measure the absorbance change value of the reaction system within 10 minutes, and the measurement wavelength is 340nm.

Embodiment 3

[0047] The determination of embodiment 3 ornithine carbamoyltransferase activity

[0048] Instrument: automatic biochemical analyzer

[0049] Determination Reagent: A two-component kit in the form of aqueous solution, the ingredients are shown in the table below:

[0050]

[0051] During the measurement, there is no fixed requirement for the ratio of reagent to sample, but it should be consistent within the same measurement batch. Here, reagent 1: sample: reagent 2 is 1800:80:600. Add the sample to be tested to reagent 1, incubate at 37°C for 5 minutes, and read the blank value of the sample, then add reagent 2, and measure after a delay of 2 minutes. Measure the absorbance change value of the reaction system within 15 minutes, and the measurement wavelength is 340nm. It is also possible to mix Reagent 1 and Reagent 2 before measurement, and then measure as a single reagent.

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Abstract

The invention discloses a detection method for activity of ornithine carbamoyltransferase (OCT). The method is characterized in that: a new cyclic increment technology is adopted for improving detection sensitivity, such that the OCT in a body fluid sample can be automatically detected as a common zymologic diagnostic reagent is adopted; the OCT in a sample, and L-ornithine, carbomoyl phosphate and citrulline hydrolase in the detection reagent, are subjected to repetitive cyclic reactions to generate carbon dioxide and ammonia; a formation rate of the ammonia is proportional to the activity of the OCT in the sample, such that the activity of the OCT in the sample can be detected through detecting the formation rate of the ammonia. According to the present invention, a full automatic zymologic detection method for the OCT, and the detection reagent thereof are firstly established; the quantitative determination reagent prepared from the method has characteristics of convenience, speed, automation and high sensitivity.

Description

technical field [0001] The invention belongs to the technical field of medical examination and measurement, and in particular relates to a method and a reagent for measuring the activity of ornithine carbamoyltransferase. Background technique [0002] Ornithine carbamoyltransferase (OCT) is an aminotransferase that is mainly located in the mitochondria of liver cells and is a liver-specific enzyme. It is most abundant in the liver, and other tissues such as the small intestine are basically absent. Its role is to catalyze the reversible transformation of ornithine to citrulline, which has important physiological functions in the body and is a key step for ammonia to generate urea through the ornithine cycle in liver cells. Determination of ornithine carbamoyltransferase is a sensitive indicator of hepatobiliary diseases. Hepatocellular carcinoma is a common malignant tumor, accounting for the third place in the mortality rate of malignant tumors. Early detection and treat...

Claims

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Application Information

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IPC IPC(8): G01N33/52G01N21/78G01N21/33C12Q1/48
Inventor 王学忠
Owner 王学忠
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