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an nh based 2 A method for the visual detection of alkaline phosphatase activity in serum of -cu-mof

A phosphatase activity, nh2-cu-mof technology, applied in the field of visual detection of alkaline phosphatase activity in serum based on NH2-Cu-MOF, can solve many influencing factors, short detection time, precision and accuracy Issues to be improved

Active Publication Date: 2020-10-20
GUANGXI NORMAL UNIV
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Problems solved by technology

However, this device requires the use of biological enzymes such as glucoamylase, and there are many factors affecting the activity of biological enzymes in the process of production, transportation and use, and only by ruler measurement and manual reading of the flow length of the solution. Still need to improve on degree and accuracy; CN107677804A a kind of bone-derived alkaline phosphatase detection kit and using method thereof, although the detection time of bone-derived alkaline phosphatase detection kit is short, but still need full-automatic biochemical analyzer to detect

Method used

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Embodiment

[0028] A NH-based 2-The method for the multicolor visualization detection of alkaline phosphatase activity in serum of Cu-MOF nanozyme, comprises the steps:

[0029] (1) Synthesis of NH with certain morphology characteristics 2 -Cu-MOF nano-enzyme material;

[0030] 1g Cu(NO 3 ) 2 ·3H 2 O was dissolved in 10 mL of deionized water; 10 mL of DMF dissolved with 0.4 g of 2-aminoterephthalic acid and 10 mL of ethanol were mixed, added to the above solution and mixed evenly; after mixing well, the solution was placed in Teflon In a lined autoclave, react at 80 °C for 24 h; finally, collect NH by centrifugation 2 -Cu-MOF, washed sequentially with DMF, ethanol and deionized water, and dried naturally.

[0031] (2) Utilize NH 2 -Cu-MOF nanozyme and agarose to prepare a gel detection tube for detecting alkaline phosphatase activity;

[0032] Mix 300 μL of Tris-HCl buffer solution with pH 7.2, 100 μL of NH 2 -The ethanol solution of Cu-MOF (2.5 g / L) was added to 1 mL of agarose ...

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Abstract

The invention discloses a method of visibly detecting alkaline phosphatase activity in serum based on NH2-Cu-MOF. The method comprises the following steps: firstly synthesizing a NH2-Cu-MOF nano-enzyme material, and preparing a gel detection tube for detecting the alkaline phosphatase activity by utilizing NH2-Cu-MOF nano-enzyme and agarose; finally adding the alkaline phosphatase in the gel detection tube, placing the gel detection tube under an ultraviolet lamp, and realizing semi-quantitative detection by observing color change; and then photographing by using a mobile phone or a digital camera, reading RGB value in a determination photo through RGB color extractor software, analyzing the RGB value to indicate oxidation degree of the OPD, thereby realizing field visual multicolor fluorescence quantitative detection of the alkaline phosphatase activity in serum; in the detection, firstly drawing a working curve that the gel color is changed along the alkaline phosphatase activity, and then detecting a serum actual sample. Through the method disclosed by the invention, the field visual multicolor fluorescence determination of the alkaline phosphatase in the serum can be realized without using an expensive instrument and equipment, the participation of the bio-enzyme, and the operation of professional.

Description

technical field [0001] The invention relates to a method for detecting alkaline phosphatase activity, specifically a method based on NH 2 -Cu-MOF visual detection method for alkaline phosphatase activity in serum. Background technique [0002] Alkaline phosphatase (ALP) is a ubiquitous hydrolase in prokaryotic and eukaryotic cells that catalyzes the removal of phosphate groups from phosphate-containing polyphosphate substrates. In the human body, alkaline phosphatase is widely present in tissues (such as bone, kidney, liver, etc.), and plays an important role in various normal cell functions such as cell signal transduction, cell division, differentiation, and bone calcification . On the other hand, the downregulation of alkaline phosphatase activity is closely related to a variety of human diseases, including hypophosphatasia, primary biliary cirrhosis, diabetes, and various cancers. Therefore, the development of a reliable and simple method for detecting alkaline phosph...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
CPCG01N21/6486
Inventor 林天然秦雨欣李金莹覃思媛黄媛林侯丽
Owner GUANGXI NORMAL UNIV
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