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Fluorescent sensor formed on the basis of DNA combining terbium ion, and application thereof for distinguishing and detecting metal ions

A technology of fluorescence sensor and metal ion, which is applied in the field of fluorescence analysis and detection, can solve the problems of in-situ real-time high-throughput rapid detection, inability to distinguish detection on a large scale, unfavorable large-scale promotion, etc., and achieve the elimination of background fluorescence and scattered fluorescence , low cost, and the effect of saving usage

Active Publication Date: 2018-08-24
EAST CHINA NORMAL UNIVERSITY
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Problems solved by technology

It overcomes the complex pre-preparation in the existing methods for distinguishing and detecting metal ions, requires expensive large-scale instruments, is difficult to achieve in-situ real-time high-throughput rapid detection, is not conducive to large-scale promotion, and requires complex probe preparation. Insufficient large-scale differential detection of metal ions

Method used

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  • Fluorescent sensor formed on the basis of DNA combining terbium ion, and application thereof for distinguishing and detecting metal ions
  • Fluorescent sensor formed on the basis of DNA combining terbium ion, and application thereof for distinguishing and detecting metal ions
  • Fluorescent sensor formed on the basis of DNA combining terbium ion, and application thereof for distinguishing and detecting metal ions

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Embodiment 1

[0035] Screened from 10 sections of DNA of different lengths, the 10μM [G 3 T] N (N=1-10) (as shown in Table 1) were added to 10 μM Tb solution and incubated for 10 min, and detected by time-resolved fluorescence analysis. like figure 2 As shown, [G 3 T] 3 , [G 3 T] 5 and [G 3 T] 6 exhibited better sensitization capabilities. These three pieces of DNA were selected as antenna ligands to sensitize Tb to form a fluorescent sensor DNA / Tb, and then the stoichiometric ratio between DNA and Tb was explored, and different concentrations of DNA ([G 3 T] 3 , [G 3 T] 5 and [G 3 T] 6 ) were added to 10 μM Tb solution to obtain different concentrations of DNA-sensitized Tb, and after incubation for 10 min, time-resolved fluorescence analysis was used for detection. see results image 3 . like image 3 As shown, [G 3 T] 3 , [G 3 T] 5 and [G 3 T] 6 Antenna sensitization ratio ([G 3 T] N / Tb) are 4:5, 2:5, 1:5 respectively. Unless otherwise specified, in the follow...

Embodiment 2

[0039] Select silver ions (Ag) and chromium ions (Cr) and blank (water) as pre-experiments, take 10 μL of different metal ion solutions to be tested, add them to the sensor solution, mix and place at room temperature for 5 minutes, and then use the method of time-resolved fluorescence analysis to test. Use the method of principal component analysis to realize the processing of fluorescence data, such as Figure 4 As shown in a-e, Ag will enhance DNA / Tb fluorescence, while Cr will cause DNA / Tb fluorescence quenching. Each group of experiments was repeated 5 times, and the fluorescence matrix was analyzed by PCA method (3DNA / Tb sensor composition × target number × 5 parallel samples), and the results were as follows Figure 4 As shown in f, these three targets (2 metals and blank) can be distinguished by the fluorescence-based sensor DNA / Tb. In order to further explore the mechanism of action, the fluorescence lifetime of DNA / Tb in these three cases was measured, as Figure 4...

Embodiment 3

[0041] In order to evaluate the discrimination ability of DNA / Tb fluorescent sensor more widely, 49 kinds of metal ions were added as targets, including alkali metals, alkaline earth metals, transition / post-transition metals and rare earth metals, and their most common or stable states were selected. Since alkali metals have a lower charge density, they are difficult to detect and distinguish by fluorescence methods, so higher concentrations are chosen. The concentration of alkali metal is 500 μM, and the concentration of other metals is 5 μM, respectively added to three DNA / Tb fluorescent sensing elements, and the detection method is as described in the above pre-experiment. The PCA analysis method was used to evaluate the metal ion-induced DNA / Tb fluorescent sensor response. like Image 6 As shown in a,c,e,f, 5 alkali metals, 4 alkaline earth metals, 24 transition / post-transition metals, and 16 rare earth metals can be clearly distinguished with high discrimination accuracy...

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Abstract

The invention discloses a fluorescent sensor formed on the basis of DNA combining terbium ion, and an application thereof for distinguishing and detecting metal ions, wherein distinguish and detectionon large-scale metal ions are achieved by means of a time resolution fluorescent method. A method includes the steps of: screening three single-chain DNAs, which are rich in guanine (G) and thymine (T) and are in different lengths, as ligands, and forming the fluorescent sensor (DNA / Tb) by sensitizing the terbium through an antenna effect; then dissolving the DNA / Tb in a buffer solution and adding 49 metal ions, comprising alkali metal ions, alkaline-earth metal ions, transition / late-transition metal ions, and rare earth metal ions, wherein the antenna effect is influenced due to interactionbetween the metal ions and DNA, so that the DNA / Tb can acquire different time resolution fluorescent signals and fluorescent service life under different metal ion conditions; finally performing principal component analysis (PCA) to analyze the fluorescent signals and fluorescent service life to achieve the distinguish and detection on metal ions. The method also achieves semi-quantitative detection on metal in river water.

Description

technical field [0001] The invention belongs to the field of fluorescence analysis and detection, and in particular relates to a fluorescence sensor combining DNA with terbium ions, which realizes the distinction and detection of large-scale metal ions by using a label-free time-resolved fluorescence and fluorescence lifetime method. Background technique [0002] The detection of biochemical species is not only the key in the field of biochemical analysis, but also plays an important role in the fields of environmental monitoring, food safety, and clinical diagnosis. For this reason, the fluorescence sensor has become an effective detection tool because of its advantages of simplicity, rapidity, real-time monitoring, simplicity, and good reproducibility. However, most fluorescent sensors suffer from interference from background fluorescence, which affects the detection sensitivity and response range. To effectively eliminate background fluorescence, the ideal method is a ti...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6408G01N21/6428G01N2021/6432
Inventor 张闽薛诗凡施国跃陈子晗韩欣悦林子杨杨苗
Owner EAST CHINA NORMAL UNIVERSITY
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