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Instant detection method for activity of alkaline phosphatase

A phosphatase activity and phosphatase technology, which is applied in the field of biochemical analysis and biosensing, can solve the problems of expensive instruments, bulky size, inability to realize on-site analysis and instant detection, etc., to reduce the cost of analysis, and the analysis process is simple and easy to implement. , to achieve the effect of on-site analysis and real-time quantitative detection

Inactive Publication Date: 2019-07-26
GUILIN UNIVERSITY OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods require high professional skills for operators, especially the instruments used are relatively expensive and bulky, resulting in high overall analysis costs, and cannot achieve on-site analysis and instant detection (Point-of-Care Testing)

Method used

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  • Instant detection method for activity of alkaline phosphatase
  • Instant detection method for activity of alkaline phosphatase
  • Instant detection method for activity of alkaline phosphatase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] 5 U / mL alkaline phosphatase (enzyme) sample solution and blank sample (blank, buffer solution without analyte) were detected by using the immediate detection method of alkaline phosphatase activity in the present invention.

[0020] Such as figure 1 As shown, the specific implementation process is as follows: step 1, add 30 µL 5 U / mL alkaline phosphatase sample solution (prepared by 20 mM tris-HCl buffer solution containing 50 mM NaCl, pH7. 2), then add 30 µL of 800 µM pyrophosphate solution (prepared from 20 mM tris-HCl buffer solution containing 50 mM NaCl, pH 7.2) to mix with it, and place it at 37 °C for 60 min; Step 2, continue to add 30 µL 400 µM Cu 2+ (copper nitrate) aqueous solution, react at room temperature for 15 min; step 3, use a pen type Cu 2+ The detection electrode measures the free Cu in the above reaction mixture solution 2+ concentration. At this time, the magnitude of the electrode response signal (Electrode signal) obtained from the sample is p...

Embodiment 2

[0023] The alkaline phosphatase sample solution with an activity concentration ranging from 0.012 to 5000 mU / mL is detected by using the immediate detection method for alkaline phosphatase activity of the present invention.

[0024] Such as figure 1 As shown, the specific steps for detecting each alkaline phosphatase sample solution in the present embodiment are: Step 1, add 30 µL of a certain activity concentration alkaline phosphatase sample solution (composed of 50 mM NaCl in 20 mM tris-HCl buffer solution, pH 7.2), then add 30 µL of 800 µM pyrophosphate solution (prepared from 50 mM NaCl in 20 mM tris-HCl buffer solution, pH 7.2) React at 37 °C for 60 min; step 2, continue to add 30 µL of 400 µM Cu 2+ (copper nitrate) aqueous solution, react at room temperature for 15 min; step 3, use a pen type Cu 2+ The detection electrode measures the free Cu in the above reaction mixture solution 2+ concentration. Plot the electrode response signal value (Electrode signal) obtained...

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Abstract

The invention discloses an instant detection method for the activity of alkaline phosphatase. Through a mechanism that Cu<2+> which is subsequently added is not subjected to complexion through the pyrophosphoric acid so as to be subjected to high-sensitivity detection through a Cu<2+> selective detection electrode after an alkaline phosphatase analyte in sample solution catalyzes pyrophosphoric acid to be subjected to hydrolysis reaction, the magnitude of an electrode response signal and the activity of alkaline phosphatase in the sample solution are in positive correlation, i.e., the instantquantitative detection of the activity of the alkaline phosphatase is finished. The method disclosed by the invention is simple and easy in implementation, and an experiment can be carried out withoutoperation personnel who do not receive professional training. In addition, large professional analysis instruments, including enzyme-labeled instruments, luminoscopes, electrochemical workstations and the like, do not need to be used for carrying out quantitative signal reading, analysis cost is greatly lowered, and the method has a wide application prospect.

Description

technical field [0001] The invention belongs to the technical field of biochemical analysis and biosensing, and in particular relates to a real-time detection method for alkaline phosphatase activity. Background technique [0002] Alkaline phosphatase, also known as alkaline phosphatase, can catalyze the hydrolysis of phosphate bonds under alkaline conditions. In mammals, alkaline phosphatase activity is relatively high in liver, bile duct, kidney, bone and placenta. Common alkaline phosphatases include intestinal alkaline phosphatase, non-tissue-specific alkaline phosphatase, and placental alkaline phosphatase. High or low alkaline phosphatase activity is associated with many diseases. For example, the activity of alkaline phosphatase in differentiated colon cancer cells will be significantly increased, which is used as a qualitative and quantitative indicator of the degree of differentiation of colon cancer cells. Elevated alkaline phosphatase in serum, known as hyperal...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26G01N27/333
CPCG01N27/26G01N27/333
Inventor 张云刘召应聂瑾芳姚茂茂
Owner GUILIN UNIVERSITY OF TECHNOLOGY
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