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Visual detection method for alkaline phosphatase detection

A detection method, phosphatase technology, applied in the field of enzyme detection, can solve the problems of time-consuming, poor stability, high cost of noble metal nanoclusters, and achieve broad application prospects and simple steps

Pending Publication Date: 2022-06-28
CHONGQING UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, organic fluorescent probes have poor water solubility, semiconductor quantum dots are highly toxic, the preparation steps of fluorescent copolymers are complicated and time-consuming, and noble metal nanoclusters are expensive and poor in stability.

Method used

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  • Visual detection method for alkaline phosphatase detection
  • Visual detection method for alkaline phosphatase detection

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Embodiment 1

[0036]Add 0.9 mL of 4-aminophenylphosphate sodium salt (25 mM) and agarose with a mass of 1% of the first mixed solution to 1.5 mL of buffer to obtain the first mixed solution; put it into a magnetic stirring electric heating mantle , after heating and stirring at 160° C. to fully melt the agarose in the first mixed solution, pour the liquid into multiple molds, each mold is filled with 240 μL of the mixed solution, and cooled at room temperature to form the first solid-state cross-linked product; 150 μL of standard alkaline phosphatase with different concentrations were added dropwise to the first solid-state cross-linked product, and heated at 70° C. for 105 min to form a second solid-state cross-linked product; each drop-wise added to the second solid-state cross-linked product 210μL of (3-aminopropyl)trimethoxysilane liquid was heated at 70°C for 80min to generate a third solid-state cross-linked product; finally, the third solid-state cross-linked product was placed under ...

Embodiment 2

[0039] Further, in the method for detecting alkaline phosphatase proposed in the present invention, step S2a can also be performed after step S1 is completed: adding a reference substance to the fourth mixed solution. These reference substances are at least one of rhodamine, fluorescent carbon quantum dots, fluorescent silicon quantum dots, and fluorescent CdTe quantum dots, and the reference substances appear red under ultraviolet light, and these reference substances react with each other under ultraviolet light. The light blue fluorescence of the product CDs showed obvious differences, which can be used as a reference to build a ratiometric fluorescence detection platform to improve the visualization effect. Most preferably, the concentration of the reference substance in the reaction system is 0.025 mg / mL.

[0040] After adding the reference substance for reaction, the third solid-state cross-linked substance was placed under the ultraviolet analyzer, and its fluorescence ...

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Abstract

The invention discloses a visual detection method for alkaline phosphatase detection, which comprises the following steps: S1, providing 4-aminobenzene sodium phosphate, agarose and a buffer solution, and adding the 4-aminobenzene sodium phosphate and the agarose into the buffer solution to obtain a first mixed solution; s2, heating and stirring to fully melt the agarose in the first mixed liquid, pouring the liquid into a mold, and cooling to form a first solid cross-linked substance; s3, adding alkaline phosphatase to be detected on the surface of the first solid-state cross-linked substance, and heating to generate a second solid-state cross-linked substance; s4, adding (3-aminopropyl) trimethoxysilane liquid on the surface of the second solid cross-linked substance, and continuously heating to generate a third solid cross-linked substance; and S5, placing the third solid cross-linked substance under an ultraviolet analyzer, and observing the fluorescence color of the third solid cross-linked substance under 365nm exciting light.

Description

technical field [0001] The present application relates to the technical field of enzyme detection, in particular to a visual detection method for alkaline phosphatase detection. Background technique [0002] Alkaline phosphatase (ALP) is an important hydrolase in mammalian tissues, which catalyzes the hydrolysis and phosphorylation of phosphate esters. As a key biomarker for medical diagnosis, ALP mainly exists in the liver, kidney, intestine, bone and serum. The concentration of ALP in the serum of healthy adults ranges from 46 to 190 mU / mL. Diseases such as worsening kidney function, liver dysfunction, cholestasis, vascular calcification, bone disease, and breast cancer. In order to protect human health, early diagnosis and detection of ALP is urgent. So far, the detection methods of ALP mainly include: colorimetry, surface-enhanced Raman scattering, electrochemical analysis and chromatography. Among these detection methods, fluorescence detection technology has receive...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428G01N21/6447
Inventor 刘玉菲韩亚琴张佳婧张国秀杨达
Owner CHONGQING UNIV
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