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Method for detecting alkaline phosphatase and cardiac troponin I in real time through in-situ fluorescence reaction initiated by copper ions and application

A cardiac troponin and fluorescent reaction technology, applied in the field of nano-bio-sensing, can solve the problems of long response time, low sensitivity, complicated steps, etc., and achieve the effects of fast response, good stability and good applicability

Active Publication Date: 2021-11-26
NANJING UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above methods have some disadvantages: complicated steps, low sensitivity and long response time

Method used

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  • Method for detecting alkaline phosphatase and cardiac troponin I in real time through in-situ fluorescence reaction initiated by copper ions and application
  • Method for detecting alkaline phosphatase and cardiac troponin I in real time through in-situ fluorescence reaction initiated by copper ions and application
  • Method for detecting alkaline phosphatase and cardiac troponin I in real time through in-situ fluorescence reaction initiated by copper ions and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1: At room temperature, take 1000 μl of Tris-HCl buffer solution, add 300 μm dopamine, 300 μm 8-hydroxygeniroli and 150 μm Cu 2+ And shake evenly. After the reaction 60s, the green fluorescence was observed in the case of ultraviolet lamp, and the HFO was successfully prepared. The DMTM is absorbed by the ultraviolet absorption spectrometer, and the absorption spectrum is drawn, such as figure 1 . Fluorescence intensity detection by fluorescence spectrometer, plotting fluorescence excitation spectrum and fluorescence emission spectrum, such as figure 2 with image 3 .

Embodiment 2

[0044] Example 2: At room temperature, 1000 μl of Tris-HCl buffer solution, 300 μm dopamine, 300 μm 8-hydroxyloli, 150 μm Cu, respectively 2+ , Shake evenly, the DMTM is scanned by the fluorescence spectrometer, the excitation wavelength is 460 nm, the emission wavelength is 520 nm, the drawing time scanning diagram, such as Figure 4 . It indicated that the generated material fluorescence intensity system was very stable.

Embodiment 3

[0045] Example 3: 300μm dopamine, 300 μm 8-hydroxy Jourli and 150 μm Cu 2+ And shake evenly. In the above system, 5 μL was added to pyrophosphate (600 μm), oscillating reaction with different concentrations of basic phosphatase (0-1250 mU / mL). The fluorescence intensity detection was performed by a fluorescence spectrometer, and the excitation wavelength was 460 nm, and the emission wavelength was 520 nm. Draw real-time fluorescence scanning drawings, fluorescence relative intensity scatter plots and standard graphs, such as Figure 5 , Image 6 with Figure 7 . It can be clearly observed to gradually increase the fluorescence intensity with the increase of ALP activity, which is very sensitive to the current experimental conditions, which can be easily detected by 0.5mu / ml of ALP.

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Abstract

The invention relates to a method for detecting alkaline phosphatase and cardiac troponin I in real time through in-situ fluorescence reaction initiated by copper ions and application. According to the method, dopamine and phenol (1, 3-naphthalene diphenol, 8-hydroxyjulolidine and 1, 5-naphthalene diphenol) are taken as raw materials, and a fluorescent compound (FCs) with high fluorescence intensity, good stability and adjustable wavelength is synthesized in a room-temperature water phase. Under the initiation of Cu <2+>, dopamine and different phenols can obtain the fluorescent substances with different emission wavelengths, and the affinity of pyrophosphate ions (PPi) to Cu <2+> is utilized to realize the fluorescence closing of a system. Under the catalytic hydrolysis of alkaline phosphatase (ALP), PPi can be quickly converted into phosphate ions, so that the fluorescence of the system is opened. A multi-channel immunofluorescence sensing platform is constructed by taking cardiac troponin I as a targeting antigen and taking alkaline phosphatase as a marker enzyme. In addition, a sensing system still shows rapid response and good recovery rate in human serum.

Description

Technical field [0001] The present invention relates to the construction of the in-situ fluorescent immunoassay sensing platform based on the wavelength of copper ion triggering and its analysis, for real-time sensitive detection CTNI, belonging to the field of nanohosphere sensing. Background technique [0002] Alkaline phosphatase (ALP) is a membrane binding enzyme, which is widely expressed in liver, bone, intestinal, placenta, kidney. It is mainly responsible for hydrolyzing and transforming monophosphate from various proteins and non-protein substrates. As a recognized biomarker, alkaline phosphatase plays an important role in many important physiological and pathological processes such as cell cycle growth, apoptosis and signal transduction pathways. The normal value of adult serum ALP is generally in 46 ~ 190u / L, children and pregnant women ALP normal value above 500u / l, but serum ALP abnormalities is close to bone disease, liver function disorders, breast cancer, pros...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N21/64
CPCG01N33/543G01N33/54386G01N21/643G01N2021/6439
Inventor 刘金华孙玉洁庞丽华常进于海东
Owner NANJING UNIV OF TECH
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