CN110878336A - Based on Fe3O4Optical sensing detection method for miRNA of @ C nanoparticles - Google Patents
Based on Fe3O4Optical sensing detection method for miRNA of @ C nanoparticles Download PDFInfo
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- CN110878336A CN110878336A CN201911125964.8A CN201911125964A CN110878336A CN 110878336 A CN110878336 A CN 110878336A CN 201911125964 A CN201911125964 A CN 201911125964A CN 110878336 A CN110878336 A CN 110878336A
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
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- C12Q1/6825—Nucleic acid detection involving sensors
Abstract
The invention belongs to the technical field of environmental monitoring, and provides a Fe-based material3O4An optical miRNA sensing detection method of @ C nanoparticles. In the invention, the design of the catalytic hairpin self-assembly coupling magnetic nanoparticles can effectively avoid the problems of nonspecific displacement and the like in the detection process, and Fe is utilized3O4The @ C nano particles have strong quenching capacity and can reduce fluorescence background signals. And under the condition of miRNA existence, the hairpin self-assembly is utilized, the signal response efficiency induced by miRNA is improved, and the specificity of miRNA detection is ensured. The fluorescence intensity of the reaction system is positively correlated with the concentration of miRNA in a certain range, thereby providing a basis for the quantitative analysis of miRNA. The detection method can effectively reduce the fluorescence background value, improve the signal response value of the target object and effectively improve the detection performance of the fluorescence sensing platform constructed based on the physical adsorption principle on the miRNA.
Description
Technical Field
The invention belongs to the technical field of environmental monitoring, and relates to a Fe-based material3O4An optical miRNA sensing detection method of @ C nanoparticles.
Background
With the continuous development of the industry in China, air, water and soil are polluted by various environments. Many environmental pollutants have the characteristic of enrichment, so that the environmental pollutants are difficult to degrade in the environment, and can be stored, accumulated and migrated after entering an ecological system, thereby causing harm, and can enter a human body through the enrichment effect to generate a toxic effect. Antibiotics, when introduced into cells of the body, stimulate the cells to produce exogenous ROS. ROS attack DNA, causing oxidative damage to DNA. This malignant lesion of the tumor is generated, so that the tumor disease marker miRNA is produced in excess. Therefore, monitoring of miRNA is an important means for prevention and treatment of diseases caused by environmental pollution. However, highly selective and sensitive detection of mirnas remains a challenging area of research due to the presence of high concentrations of other interfering nucleic acid components in organisms.
The conventional miRNA detection methods, such as a PCR method, a quantitative PCR method and the like, generally have the problems of poor anti-interference capability, complex early operation and the like, so that the conventional miRNA detection methods are not suitable for real-time online continuous detection. In recent years, the biosensing technology has many advantages that the traditional miRNA detection technology does not have, such as portability, simple instrumentation, relatively low detection cost, and rapid development. In addition, the method also has the advantages of good sensitivity, wide linear range, strong anti-interference capability and the like, and more importantly, the method provides possibility for realizing automatic real-time online monitoring.
In recent years, a sensing method constructed based on a nano material is taken as a novel analysis method, has the advantages of simplicity and convenience in operation, high sensitivity, good specificity, high response speed and the like, and shows a wide application prospect in the analysis field. Researchers have successfully constructed a series of sensing platforms for miRNA detection (anal. chem.2016,88, 4254-4258; anal. chem.2014,86,2124-2130) by using nanomaterials, and the signal response mode comprises electrochemistry, photoelectricity, optics and the like. Among them, the optical detection method has become an analysis method with great application potential due to simple equipment, easy miniaturization and high sensitivity. At present, various optical sensors are researched and used for detecting miRNA and obtain good detection effect (anal. chem.2019,91, 3374-3381; ACS apple. mater. interfaces 2014,6, 1152-1157). The basic principle of the fluorescence sensing method is to utilize the Fluorescence Resonance Energy Transfer (FRET) function between a fluorophore and a quencher along with the change of distance, and further realize the detection of a target through the change of a fluorescence signal (Angewandte Chemie-International Edition,2009,48(26): 4785-4787). However, the fluorescence sensing methods constructed based on the physical adsorption principle have inherent defects of nonspecific displacement, low desorption efficiency and the like, and influence the detection sensitivity of miRNA to a certain extent. In recent years, DNA nanotechnology based on catalytic hairpin self-assembly has been widely used in the construction of fluorescence sensing methods because of its ability to achieve signal amplification without enzyme-assisted conditions (anal. chem.2015.87(10): 5430-6.). However, due to the low quenching efficiency of most quenching probes, the fluorescence sensor still has relatively high fluorescence background. Therefore, it is of great significance to develop a new low-background high-response optical sensing method for detecting miRNA.
Magnetic nanoparticles are a class of nano materials mainly composed of metal oxides and having a magnetic effect, and have the advantages of strong capability of separating substances, simple separation process, large specific surface area and the like, so that the magnetic nanoparticles have great application potential in the field of sensing (anal. chem.2016,88, 4254-. In order to avoid the inherent defects of the physical adsorption type fluorescence sensing method, the invention utilizes the emerging Fe3O4The @ C nano-particle is designed with a catalytic hairpin self-assembly covalent coupling magnetic nano-particle-based fluorescence sensing method, so that the defects of high background signal, low desorption efficiency and the like of the traditional physical adsorption type fluorescence sensing method are effectively overcome, the signal response degree induced by a target object is enhanced, the detection performance of the target object is improved, and the method is simple to operate, strong in practicability and wide in application prospect. Thus, the present invention utilizes catalytic hairpin self-assembly covalent coupling of Fe3O4The @ C nano-particles realize the optical sensing detection of miRNA, and the sensor pair prepared by the methodThe kit has the advantages of quick response, high sensitivity and selectivity and wide application prospect in miRNA detection.
Disclosure of Invention
The invention solves the defects of poor specificity, time consumption, poor anti-interference performance, high background signal, weak response signal and the like of the traditional method, and provides a simple, rapid and high-selectivity method for accurately detecting miRNA.
In the invention, three nucleic acid sequences are designed in the design of the catalytic hairpin self-assembly coupled magnetic nanoparticles, wherein a signal molecule can be covalently coupled to Fe through a hybrid amino-modified nucleic acid molecule3O4On the @ C nano-particles, the problems of nonspecific displacement and the like in the sensing detection process can be effectively avoided, and Fe is utilized3O4The @ C nano particles have strong quenching capacity, and the background signal of the physical adsorption type fluorescence sensing method is reduced. And under the condition of miRNA existence, the signal response efficiency induced by miRNA is improved by the circulating amplification of catalytic hairpin self-assembly. The fluorescence intensity of the reaction system is positively correlated with the concentration of miRNA in a certain range, thereby providing a basis for the quantitative analysis of miRNA. The designed signal molecule has specific recognition capability, and can ensure the specificity of miRNA detection.
The technical scheme of the invention is as follows:
1. based on Fe3O4The optical miRNA sensing detection method of the @ C nano-particles is characterized in that the steps of the design of catalytic hairpin self-assembly and coupling magnetic nano-particles are as follows:
(1) designing a nucleic acid sequence H1 containing partial complementarity of miRNA as a main body; designing a nucleic acid sequence H3 and covalently coupling the nucleic acid sequence H3 to the magnetic nanoparticles to form a quenching probe; designing a nucleic acid sequence H2 with fluorescent modification as a signal molecule, wherein a partial sequence of the signal molecule is complementary with a partial sequence of the nucleic acid sequence H1, a partial sequence of the signal molecule is complementary with H3, and after the signal molecule is complementary, the fluorescence can be quenched by a quenching probe;
(2) mixing a nucleic acid sequence H1 with miRNAs with different concentrations, and incubating at 5-60 ℃ until the hybridization reaction is completed;
(3) mixing the signal molecule H2 and the quenching probe until the hybridization reaction is completed;
(4) mixing the step (2) and the step (3) to complete competitive hybridization reaction;
(5) sticking the magnet substances to the outer wall of the vessel filled with the solution reacted in the step (4), realizing solid-liquid separation, and obtaining a supernatant solution; and measuring the fluorescence intensity of the supernatant solution to finish the extracellular detection.
2. Based on Fe3O4The optical miRNA sensing detection method of the @ C nano-particles is characterized in that the steps of the design of catalytic hairpin self-assembly and coupling magnetic nano-particles are as follows:
(1) designing a nucleic acid sequence H1 containing partial complementarity of miRNA as a main body; designing a nucleic acid sequence H3 and covalently coupling the nucleic acid sequence H3 to the magnetic nanoparticles to form a quenching probe; designing a nucleic acid sequence H2 with fluorescent modification as a signal molecule, wherein a partial sequence of the signal molecule is complementary with a partial sequence of the nucleic acid sequence H1, a partial sequence of the signal molecule is complementary with H3, and after the signal molecule is complementary, the fluorescence can be quenched by a quenching probe
(2) Mixing a signal molecule H2 and a quenching probe at the temperature of 5-60 ℃ until the hybridization reaction is completed;
(3) mixing the solution after the hybridization reaction in the step (2) with a nucleic acid sequence H1, transfecting the solution and the nucleic acid sequence H1 into cells together, washing off the untransfected solution, and continuing to incubate the cells;
(4) and (4) observing the change of the fluorescence intensity of the cells in the step (3) along with time to finish the intracellular detection.
The invention has the beneficial effects that:
(1) the fluorescence background value is reduced by utilizing the strong quenching capacity of the carbon material and through a covalent coupling nucleic acid mode.
(2) The magnetic nano particles are separated from the signal molecules, so that the desorption of the signal molecules is promoted, and the fluorescence recovery efficiency is improved.
(3) By utilizing the cyclic amplification of the catalytic hairpin self-assembly, the fluorescence recovery efficiency is enhanced, and the detection performance of miRNA is improved.
(4) The fluorescence sensor realizes high sensitivity and high selectivity detection of miRNA in vitro and successfully realizes practical detection application in human cells.
Drawings
FIG. 1 is a Fe-based alloy according to the present invention3O4The design process and the detection mechanism of the miRNA optical sensing detection method of the @ C nano-particles are schematically shown.
Legend:
FIG. 2 is a standard working curve of the nano-optical sensor obtained by the method of the present invention applied to miRNA.
Detailed Description
The following detailed description of the embodiments of the invention is provided in connection with the accompanying drawings and the detailed description of the embodiments.
Example 1
Determination of miRNA in the prepared water sample:
(1) design of catalytic hairpin self-assembling nucleic acids: h1 is designed as a main body, H2 is used as a signal molecule, and H3 modified amino is covalently coupled to Fe3O4@ C nanoparticle surface, forming quenching probes. Respectively diluting substrate chains H1, H2 and H3 with different sequences to 43 μ M, 150nM and 100nM, respectively placing in a water bath, heating to 95 deg.C, maintaining for 5-10min, and naturally cooling to room temperature;
(2) preparation of quenching probe: fe3O4Mixing the @ C nano particles and EDC at 4-50 ℃ for 20 min; further adding NHS, wherein Fe3O4The proportion of the mass of the @ C nano particles, the mole of EDC and the mole of NHS is 0.02g to 0.0004 to 0.0001; mixing, and adding Fe at 4-50 deg.C3O4@ C nanoparticle surface group activation for 1 h; activated Fe3O4Mixing the @ C nanoparticles with H3, incubating for 24H at 4-50 deg.C to form covalent coupling to form quenching probe (H3-Fe)3O4@ C); wherein the activated Fe3O4The molar ratio of the mass of the @ C nanoparticles to H3 was 0.02g:0.000000043 mol;
(3) binding of signal molecule to quenching probe: under the temperature condition of 20-37 ℃, the fluorescence modified H2 is continuously reacted with H3-Fe3O4@ C incubated together for 24H to form H2/H3-Fe3O4@ C, where H3-Fe3O4The molar ratio of the mass of @ C to H2 is 0.001g:0.0000000001875mol, washed three times with water and left as substrate H2/H3-Fe3O4@ C plus Bis-Tris buffer and freezing for preservation at-4 to-20 ℃;
(4) in-vitro detection of miRNA: mixing H2/H3-Fe3O4@ C, H1 is mixed with miRNA buffer solution to be tested, wherein H3-Fe3O4The mass of @ C and the molar ratio of H1 are 0.000037g:0.000000000005mol, the mixture is transferred to a quartz cuvette after reacting for 2-4H at 25-37 ℃, water is added, and the change curve of the fluorescence intensity of the system along with the emission wavelength is recorded;
the nucleic acid design is as follows:
(5) drawing a standard working curve:
in the step (4), the fluorescence intensity of the reaction system at the fluorescent group absorption peak is continuously increased along with the increase of the miRNA concentration in the sample, the fluorescence intensity of the reaction system has a good linear relation with the miRNA concentration in the range of 225fM to 225pM, and the linear correlation coefficient R20.99 (fig. 2).
Example 2
Determination of intracellular mirnas:
(1) design of catalytic hairpin self-assembling nucleic acids: h1 is designed as a main body, H2 is used as a signal molecule, and H3 modified amino is covalently coupled to Fe3O4@ C nanoparticle surface, forming quenching probes. Respectively diluting substrate chains H1, H2 and H3 with different sequences to 43 μ M, 150nM and 100nM, respectively placing in a water bath, heating to 95 deg.C, maintaining for 5-10min, and naturally cooling to room temperature.
(2) Preparation of quenching probe: fe3O4Mixing the @ C nano particles and EDC at 4-50 ℃ for 20 min; further adding NHS, wherein Fe3O4The proportion of the mass of the @ C nano particles, the mole of EDC and the mole of NHS is 0.02g to 0.0004 to 0.0001; mixing, and adding Fe at 4-50 deg.C3O4@ C nanoparticle tableActivating a surface group for 1 h; activated Fe3O4Mixing the @ C nanoparticles with H3, incubating for 24H at 4-50 deg.C to form covalent coupling to form quenching probe (H3-Fe)3O4@ C); wherein the activated Fe3O4The molar ratio of the mass of the @ C nanoparticles to H3 was 0.02g:0.000000043 mol;
(3) binding of signal molecule to quenching probe: at the temperature of 20-37 ℃, the fluorescence modified H2 is continuously reacted with H3-Fe3O4@ C incubated together for 24H to form H2/H3-Fe3O4@ C, where H3-Fe3O4The molar ratio of the mass of @ C to H2 is 0.001g:0.0000000001875mol, washed three times with water and left as substrate H2/H3-Fe3O4@ C plus Bis-Tris buffer and freezing for preservation at-4 to-20 ℃;
(4) and (3) miRNA intracellular detection: mixing H2/H3-Fe3O4@ C, H1 was incubated with the cells for 4h, washed three times with PBS buffer, and the cells were cultured and observed for changes in fluorescence intensity over time;
the nucleic acid design is as follows:
(5) observing the change of the fluorescence intensity of the cells;
in the step (4), the fluorescence intensity of the reaction system is continuously increased along with the increase of time of the sample in 0-24 h.
Claims (2)
1. Based on Fe3O4The optical miRNA sensing detection method of the @ C nano-particles is characterized in that the steps of the design of catalytic hairpin self-assembly and coupling magnetic nano-particles are as follows:
(1) designing a nucleic acid sequence H1 containing partial complementarity of miRNA as a main body; designing a nucleic acid sequence H3 and covalently coupling the nucleic acid sequence H3 to the magnetic nanoparticles to form a quenching probe; designing a nucleic acid sequence H2 with fluorescent modification as a signal molecule, wherein a partial sequence of the signal molecule is complementary with a partial sequence of the nucleic acid sequence H1, a partial sequence of the signal molecule is complementary with H3, and after the signal molecule is complementary, the fluorescence can be quenched by a quenching probe;
(2) mixing a nucleic acid sequence H1 with miRNAs with different concentrations, and incubating at 5-60 ℃ until the hybridization reaction is completed;
(3) mixing the signal molecule H2 and the quenching probe until the hybridization reaction is completed;
(4) mixing the step (2) and the step (3) to complete competitive hybridization reaction;
(5) sticking the magnet substances to the outer wall of the vessel filled with the solution reacted in the step (4), realizing solid-liquid separation, and obtaining a supernatant solution; and measuring the fluorescence intensity of the supernatant solution to finish the extracellular detection.
2. Based on Fe3O4The optical miRNA sensing detection method of the @ C nano-particles is characterized in that the steps of the design of catalytic hairpin self-assembly and coupling magnetic nano-particles are as follows:
(1) designing a nucleic acid sequence H1 containing partial complementarity of miRNA as a main body; designing a nucleic acid sequence H3 and covalently coupling the nucleic acid sequence H3 to the magnetic nanoparticles to form a quenching probe; designing a nucleic acid sequence H2 with fluorescent modification as a signal molecule, wherein a partial sequence of the signal molecule is complementary with a partial sequence of the nucleic acid sequence H1, a partial sequence of the signal molecule is complementary with H3, and after the signal molecule is complementary, the fluorescence can be quenched by a quenching probe
(2) Mixing a signal molecule H2 and a quenching probe at the temperature of 5-60 ℃ until the hybridization reaction is completed;
(3) mixing the solution after the hybridization reaction in the step (2) with a nucleic acid sequence H1, transfecting the solution and the nucleic acid sequence H1 into cells together, washing off the untransfected solution, and continuing to incubate the cells;
(4) and (4) observing the change of the fluorescence intensity of the cells in the step (3) along with time to finish the intracellular detection.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040067503A1 (en) * | 2002-04-22 | 2004-04-08 | Weihong Tan | Functionalized nanoparticles and methods of use |
| US20150218662A1 (en) * | 2012-08-30 | 2015-08-06 | Southwest Hospital Of Chongquing | Method for detecting and typing nucleic acids of pathogenic microorganism without amplification |
| CN109486906A (en) * | 2018-11-16 | 2019-03-19 | 中山大学附属第五医院 | MicroRNA non-enzymatic amplification detection method intracellular based on the affine nanometer transport vehicle of electrostatic and cell imaging |
| CN110095608A (en) * | 2019-04-12 | 2019-08-06 | 南方医科大学南方医院 | Tumour excretion body nano fluorescent sensor based on Magnetic Isolation and DNA self assembly |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040067503A1 (en) * | 2002-04-22 | 2004-04-08 | Weihong Tan | Functionalized nanoparticles and methods of use |
| US20150218662A1 (en) * | 2012-08-30 | 2015-08-06 | Southwest Hospital Of Chongquing | Method for detecting and typing nucleic acids of pathogenic microorganism without amplification |
| CN109486906A (en) * | 2018-11-16 | 2019-03-19 | 中山大学附属第五医院 | MicroRNA non-enzymatic amplification detection method intracellular based on the affine nanometer transport vehicle of electrostatic and cell imaging |
| CN110095608A (en) * | 2019-04-12 | 2019-08-06 | 南方医科大学南方医院 | Tumour excretion body nano fluorescent sensor based on Magnetic Isolation and DNA self assembly |
Non-Patent Citations (4)
| Title |
|---|
| DWIGHT S. SEFEROS等: "Nano-Flares: Probes for Transfection and mRNA Detection in Living Cells", 《J. AM. CHEM. SOC.》 * |
| JIANYUAN DAI等: "Rapid DNA detection based on self-replicating catalyzed hairpin assembly using nucleotide base analog pyrrolo-deoxycytidine as fluorophore", 《TALANTA》 * |
| 杨立敏等: "纳米荧光探针用于核酸分子的检测及成像研究", 《化学学报》 * |
| 郑爱仙等: "核酸功能化纳米探针在细胞荧光成像中的应用", 《中国光学》 * |
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