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a fe-based 3 o 4 @c Nanoparticle miRNA optical sensing detection method
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A detection method and optical sensing technology, which are applied in biochemical equipment and methods, and the determination/inspection of microorganisms to achieve high sensitivity, enhance fluorescence recovery efficiency, and improve detection performance.
Active Publication Date: 2022-06-14
DALIAN UNIV OF TECH
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[0006]The present invention solves the shortcomings of traditional methods such as poor specificity, time-consuming, poor anti-interference, high background signal and weak response signal, and provides a simple, fast, high-efficiency Selective and accurate method for miRNA detection
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Embodiment 1
[0030] Configure the determination of miRNA in water samples:
[0031] (1) Design of catalytic hairpin self-assembled nucleic acid: H1 is designed as the main body, H2 is used as the signal molecule, H3 modified amino group is covalently coupled to Fe 3 o 4 @C Nanoparticle surface, forming quenching probes. Dilute the substrate chains H1, H2 and H3 with different sequences to 43μM, 150nM and 100nM, respectively, put them in a water bath, heat to 95°C, keep for 5-10min, and then cool naturally to room temperature;
[0032] (2) Preparation of quenching probe: Fe 3 o 4 Mix @C nanoparticles and EDC at 4-50°C for 20min; then add NHS, in which Fe 3 o 4 The ratio of the mass of @C nanoparticles, the mole of EDC and the mole of NHS is 0.02g:0.0004mol:0.0001mol; after mixing, under the condition of 4-50℃, Fe 3 o 4 Activation of @C nanoparticle surface groups for 1 h; activated Fe 3 o 4 Mix @C nanoparticles with H3 and incubate at 4-50°C for 24h to form a covalent coupling to f...
Embodiment 2
[0040] Determination of intracellular miRNA:
[0041] (1) Design of catalytic hairpin self-assembled nucleic acid: H1 is designed as the main body, H2 is used as the signal molecule, H3 modified amino group is covalently coupled to Fe 3 o 4 @C Nanoparticle surface, forming quenching probes. Dilute the substrate chains H1, H2 and H3 with different sequences to 43μM, 150nM and 100nM respectively, put them in a water bath, heat to 95°C, keep for 5-10min and then cool to room temperature naturally.
[0042] (2) Preparation of quenching probe: Fe 3 o 4 Mix @C nanoparticles and EDC at 4-50°C for 20min; then add NHS, in which Fe 3 o 4 The ratio of the mass of @C nanoparticles, the mole of EDC and the mole of NHS is 0.02g:0.0004mol:0.0001mol; after mixing, under the condition of 4-50℃, Fe 3 o 4 Activation of @C nanoparticle surface groups for 1 h; activated Fe 3 o 4 Mix @C nanoparticles with H3 and incubate at 4-50°C for 24h to form a covalent coupling to form a quenching pro...
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Abstract
The invention belongs to the technical field of environmental monitoring and provides a Fe-based 3 o 4 Optical sensing and detection method of miRNA by @C nanoparticles. In the present invention, the design of catalytic hairpin self-assembly coupled magnetic nanoparticles can effectively avoid problems such as non-specific replacement in the detection process, and utilize Fe 3 o 4 The strong quenching ability of @C nanoparticles can reduce the fluorescence background signal. And in the presence of miRNA, the hairpin self-assembly is used to improve the signal response efficiency induced by miRNA and ensure the specificity applied to miRNA detection. The fluorescence intensity of the reaction system is positively correlated with the concentration of miRNA within a certain range, thus providing a basis for the quantitative analysis of miRNA. The detection method of the present invention can effectively reduce the fluorescence background value, increase the signal response value of the target object, and effectively improve the detection performance of the miRNA by the fluorescence sensing platform constructed based on the principle of physical adsorption.
Description
technical field [0001] The invention belongs to the technical field of environmental monitoring and relates to a Fe-based 3 o 4 Optical sensing and detection method of miRNA by @C nanoparticles. Background technique [0002] With the continuous development of my country's industry, air, water, and soil have all been polluted by various environments. Many environmental pollutants have the characteristics of enrichment, making it difficult to be degraded in the environment. After entering the ecosystem, they will persist, accumulate and migrate, causing harm, and can also enter the human body through enrichment to produce toxic effects. After antibiotics enter the cells of the body, they will stimulate the cells to produce exogenous ROS. ROS attack DNA, causing DNA oxidative damage. The malignant lesion of tumor produces excessive production of tumor disease marker miRNA. Therefore, monitoring miRNA is an important means for the prevention and treatment of diseases caused...
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