Target molecule detection method based on nanometer-gold and nucleic acid structure and kit thereof

A detection method and technology of target molecules, which are applied in the direction of chemical reaction of materials for analysis, chemiluminescence/bioluminescence, etc., can solve the problems of complicated operation and limited sensitivity, and achieve the effects of high sensitivity, improved sensitivity and rapid detection.

Inactive Publication Date: 2010-09-15
苏州市长三角系统生物交叉科学研究院有限公司
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  • Abstract
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Problems solved by technology

[0009] The technical problem to be solved by the present invention is to overcome the defects of different design of detection schemes, complicated operation and limited sensitivity for different target molecules in the existing colorimetric method for detecting target DNA of nano gold, and provide a simple and fast , general-purpose, high-sensitivity colorimetric method for detecting target substances with gold nanoparticles and its kit, which can easily detect any target substance

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  • Target molecule detection method based on nanometer-gold and nucleic acid structure and kit thereof
  • Target molecule detection method based on nanometer-gold and nucleic acid structure and kit thereof
  • Target molecule detection method based on nanometer-gold and nucleic acid structure and kit thereof

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Embodiment 1

[0040] The detection of embodiment 1 cocaine

[0041] Steps: take 2 μL cocaine-nucleic acid aptamer solution with a concentration of 100 μM and 2 μL aqueous solution of cocaine hydrochloride with a concentration of 1-100 μM, add 16 μL buffer solution (25mM Tris, pH8.2, 0.3M NaCl) and mix well, React at room temperature for 10 minutes. At the same time, no cocaine was added, 2 μL of water was added as a blank group, and two groups of 2 μL of 1 mM benzoylecgonine (N1) and ecgonine methyl ester (N2) were added as controls. Then take 2 μL of the above-mentioned reaction solution and add 100 μL of the nano-gold solution (13nm, 3.5nM) prepared above, and react at room temperature for 5 minutes, then add the above buffer solution (25mM Tris, pH8.2, 0.3M NaCl) to make up 0.2mL Record the fluorescence spectra of the experimental group and the control group.

[0042] Result: See the fluorescence spectrum figure 2 , it can be seen that in a group of solutions containing cocaine, the ...

Embodiment 2

[0043] The detection of embodiment 2ATP

[0044] Steps: Take 2 μL of ATP-nucleic acid aptamer solution with a concentration of 100 μM and 2 μL of an aqueous solution of ATP with a concentration of 0.1 μM to 100 mM, add 16 μL of buffer solution (25mM Tris, pH 8.2, 0.15M NaCl) and mix well, at room temperature Under reaction 30min. At the same time, a group without adding ATP and adding 2 μL of water was used as a blank, and a group of adding 2 μL of 1 mM CTP, UTP, and GTP was used as a control. Then take 2 μL of the above-mentioned reaction solution and add 100 μL of the above-prepared gold nano-gold solution (20nm, 1nM), react at room temperature for 5 minutes, add the above-mentioned buffer solution to make up 0.2mL, and record the fluorescence spectra of the experimental group and the control group.

[0045] Result: according to figure 1 According to the principle and operation steps shown, the fluorescence spectrum showed that the fluorescence of the group added with ATP ...

Embodiment 3

[0046] Embodiment 3 is to the detection of divalent mercury ion

[0047] Step: Take 1 μL of Hg with a concentration of 1 μM 2+ -MSO solution with 1 μL of Hg at a concentration of 10 nM to 0.1 mM 2+ Add 18 μL of water, mix well, and react for 1 min at room temperature. without adding Hg 2+ , add 1 μL of water to a group as a blank experiment. In addition, selectivity analysis was carried out through two groups of experiments, one group added 2 μL of 25 mM Ca 2+ , Mg 2+ The other group added 0.5mM mixed ions (Fe 2+ , Cu 2+ ,Co 2+ , Mn 2+ , Ni 2+ , Zn 2+ , Cd 2+ ). Then take 5 μL of the above-mentioned reaction solution and add 5 μL of the nano-gold solution (10nm, 200nM) prepared above, and react at room temperature for 5 minutes, add 0.034% citric acid solution (to prevent the nano-gold from being unstable and causing aggregation and discoloration) to make up 0.2mL Then record the fluorescence spectra of the experimental group and the control group.

[0048] Resul...

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Abstract

The invention relates to a target molecule detection method based on nanometer-gold and nucleic acid structure and a kit thereof. The target molecule detection method comprises the following steps of: (1) reacting specific DNA which can be specifically combined with the target molecules with a solution to be detected; (2) mixing the reaction solution obtained in the step (1) with a nanometer-gold solution for reaction; and (3) observing the fluorescence spectrum of the reaction solution finished in the step (2). The detection method has generality, the detected object can be an arbitrary target molecule, and the application range is wide. By utilizing the invention, the target molecules can be fast, sensitively and selectively detected.

Description

technical field [0001] The invention particularly relates to a target molecule detection method based on nano gold and nucleic acid structure and a kit thereof. Background technique [0002] Nucleic acid aptamer (aptamer) refers to the short single-stranded oligonucleotide pairing obtained by screening from a random oligonucleotide library using the SELEX (systematic evolution of ligands by exponential enrichment) in vitro screening technology established at the end of the last century. group, it can specifically bind to the target molecule, thereby undergoing a conformational change itself. [0003] Through in vitro screening technology, nucleic acid aptamers of any substance can theoretically be screened, coupled with the technical characteristics of high-throughput screening and the characteristics of precise recognition of nucleic acid aptamers, easy in vitro synthesis and modification, etc., making nucleic acid aptamers in analytical chemistry and It has broad applicat...

Claims

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Application Information

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IPC IPC(8): G01N21/76
Inventor 樊春海王丽华宋世平
Owner 苏州市长三角系统生物交叉科学研究院有限公司
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