An electrochemical sensor for rapid detection of unstable substances and a method for rapid quantitative detection of ATP using the same
An electrochemical and unstable technology, applied in the direction of material electrochemical variables, measuring devices, scientific instruments, etc., can solve problems such as low accuracy and unstable properties, and achieve accelerated molecular diffusion, high-sensitivity content detection, and biocompatibility The effect of improving the conductivity and conductivity
Active Publication Date: 2020-10-13
SHAANXI NORMAL UNIV
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Problems solved by technology
[0005] At present, although the use of biosensors based on ATP nucleic acid aptamers to detect ATP has been reported (J.AM.CHEM.SOC.2007, 129, 1042-1043.), the use of conventional large electrodes compatible with ATP nucleic acid The biosensor prepared by ligand assembly needs to hybridize with ATP for several hours when used for ATP detection, and in view of the instability of ATP's own properties, for the conventional ATP detection method that needs to hybridize with ATP for several hours, less accurate
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Embodiment 1
[0037] The method for rapid quantitative detection of ATP of the present invention is realized by the following steps:
[0038] (1) Mix anti-ATP aptamers with a concentration of 1 μmol / L and their complementary strands into tris(2-carboxyethyl)phosphine (abbreviated as TCEP, 98+%, purchased from Sigma) containing 1 mmol / L and 1mol / L NaClO 4 , in a phosphate buffer solution with a pH of 7.4, heated to 80°C, and slowly cooled to room temperature to obtain dsDNA;
[0039] The anti-ATP aptamer and its complementary chain were synthesized by Shanghai Sangon Bioengineering Co., Ltd.
[0040] The sequence of the anti-ATP aptamer is:
[0041] 5'-ferrocene-C6-ACCTGGGGGAGTATTGCGGAGGAAGGTTT-C6-SH-3';
[0042] The sequence of the complementary strand is:
[0043] 5'-ACCTTCC TCCGC AATAC TCCCC CAGGT-3'.
[0044] (2) Ultrasonically clean the Pt disk ultramicroelectrode with ultrapure water for 5 minutes, use the Pt disk ultramicroelectrode as the working electrode, and put in a solution...
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Abstract
The invention relates to an electrochemical sensor used for rapid detection of non-stable substances, and a method used for rapid quantitative determination of ATP using the electrochemical sensor. According to a preparation method, the surface of an ultramicro electrode is coated with a layer of nanoflower formed via Au nanometer particle accumulation through electrodeposition, so that an Au nanoflower ultramicro electrode is obtained; and then, assembling of double-chain DNA (dsDNA) onto the surface of the Au nanoflower ultramicro electrode is carried out. In quantitative determination of ATP, the electrochemical sensor is subjected to post-treatment so as to blocking active sites on the electrode surface; hybridization reaction of the Au nanoflower electrode assembled with dsDNA is carried out in an ATP solution so as to unblock the active sites on the electrode surface; square wave voltammetry can be adopted for rapid quantitative determination of ATP; rapid, accurate determinationon substances with unstable properties is realized; detection results are accurate; reaction sensitivity is high; and problems in the prior art such as long time consumption and large detection result error are solved.
Description
technical field [0001] The invention belongs to the technical field of ATP detection, in particular to an electrochemical sensor for rapidly detecting unstable substances and a method for rapidly and quantitatively detecting ATP based on the electrochemical sensor. Background technique [0002] ATP (adenosine triphosphate) is an unstable high-energy compound composed of one molecule of adenine, one molecule of ribose and three molecules of phosphate groups. It can release a large amount of energy when hydrolyzed, and is the most direct energy source in the body. As the main carrier of chemical energy in organisms, ATP is an important substrate in organisms and plays an important role in the regulation and integration of cell metabolism. Due to its ubiquity in organisms, it is also used as an indicator of cell survival and cell damage. Because the level of ATP in cells is also related to many diseases, such as cardiovascular disease, cardiomyopathy and Parkinson's disease, e...
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IPC IPC(8): G01N27/30G01N27/327G01N27/48
CPCG01N27/30G01N27/327G01N27/48
Inventor 郭志慧唐丽芳郑行望
Owner SHAANXI NORMAL UNIV