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Self-energized nucleic acid aptamer sensor and preparation method and application thereof

A nucleic acid aptamer and sensor technology, which is applied in the field of electrochemical analysis and environmental detection, can solve the problems of limiting ATZ rapid and sensitive detection, time-consuming analysis, and high requirements for analysts, so as to improve selective recognition ability and high sensitivity The effect of recognition ability and improvement of detection efficiency

Active Publication Date: 2018-10-12
TONGJI UNIV
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Problems solved by technology

[0003] At present, the detection methods for ATZ mainly include high performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LC-MS), capillary electrophoresis, protein phosphatase inhibition (PPIA), etc. Although these methods have high sensitivity, they are The shortcomings of expensive instruments, complicated sample pretreatment, time-consuming analysis, and high requirements for analysts limit the rapid and sensitive detection of ATZ.

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  • Self-energized nucleic acid aptamer sensor and preparation method and application thereof

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Embodiment 1

[0039] An ATZ detection sensor based on bio-enzyme fuel cell self-powered sensing and nucleic acid aptamer sensing, its negative and positive electrodes are obtained through the following steps:

[0040] 1) Pretreatment of multi-walled carbon nanotubes (CNTs): Weigh about 0.05g of CNTs in 50mL 3mol / L nitric acid (HNO 3 ) solution, placed in a round bottom flask, condensed and refluxed in an oil bath at 110°C for 8h. Take it out and wash it with distilled water. During the process, use a high-speed centrifuge (14000r / min) to separate the CNTs and washing liquid until they are neutral, and then dry them in an oven at 70°C.

[0041] 2) Preparation of GCE / CNTs-COOH / GDH catalytic anode: take out about 0.08 g of dried and pretreated CNTs obtained in step 1), add 10 mL of absolute ethanol solution, and sonicate for 15 minutes, so that the CNTs are evenly dispersed; Take 5 μL of the dispersion liquid and drop it on the surface of the GCE substrate electrode, and dry it in the air for...

Embodiment 2

[0050] A self-powered nucleic acid aptamer sensor, the sensor includes an anode electrode and a cathode electrode, the anode electrode is GCE / CNTs-COOH / GDH for spontaneous catalytic oxidation reaction, and the cathode electrode is Au / Apt;

[0051] Among them, GCE / CNTs-COOH / GDH is a glassy carbon electrode modified by carboxylated multi-walled carbon nanotubes loaded with glucose dehydrogenase, and Au / Apt is a gold electrode loaded with nucleic acid aptamer.

[0052] The sensor also includes an anolyte, a catholyte, and a proton exchange membrane disposed between the anolyte and the catholyte. The anolyte is a phosphate buffer solution containing glucose and reduced nicotinamide adenine dinucleotide, and the catholyte is a solution containing K 3 [Fe(CN) 6 ] in phosphate buffered saline.

[0053] The preparation method of the self-powered nucleic acid aptamer sensor comprises the following steps:

[0054] 1) Pretreatment of CNTs: Add CNTs to nitric acid solution, then condense...

Embodiment 3

[0062] In this embodiment, the pretreatment process of CNTs is as follows: adding CNTs to nitric acid solution, then condensing and refluxing at 100° C. for 10 h, and obtaining CNTs-COOH after washing.

[0063] Interfering substances include glyphosate, trichlorfon and monosultap.

[0064] All the other are with embodiment 2.

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Abstract

The invention relates to a self-energized nucleic acid aptamer sensor and a preparation method and an application thereof. The sensor comprises a positive electrode and a negative electrode, the positive electrode is used for a spontaneous catalytic oxidation reaction of GCE / CNTs-COOH / GDH, and the negative electrode is Au / Apt; the preparation method of the sensor comprises the following steps: CNTs pretreatment, preparation of GCE / CNTs-COOH / GDH, and preparation of Au / Apt; and the sensor is used for detecting atrazine in liquid. Compared with the prior art, a self-energized sensing analysis method with simple operation and rapid detection as well as a nucleic acid aptamer technology having specific recognition effect on ATZ are combined, the invention provides a method for construction of the self-energized nucleic acid aptamer sensor based on a bio-enzyme fuel cell as well as high-sensitivity high-selectivity analysis detection method on pesticide atrazine in a water body, the detection sensitivity can achieve 7.5*10<-9> mol / L, and the sensor has good reappearance, the sensor has excellent selectivity identification in a hundredfold-concentration hinterferent-existed system, and the problems of low sensitivity, long analysis time consuming and complex operation in the current ATZ detection technology can be solved.

Description

technical field [0001] The invention belongs to the technical field of electrochemical analysis and environmental detection, and relates to a self-powered nucleic acid aptamer sensor and its preparation method and application. Background technique [0002] In recent years, the harmful effects of environmental endocrine disruptors on humans and wildlife have become an international hot issue. Among them, there are 44 kinds of pesticide endocrine disruptors, accounting for almost two-thirds of endocrine disruptors. The common triazine herbicide Atrazine (ATZ) has a large dosage, long residual time, and good water solubility. It is very easy to cause pollution to soil, crops, and surface water when used. Discharge into the sea will also cause damage to the marine environment and ecology. serious hazard. In 1986, the European Union restricted the concentration of atrazine pesticides and their degradation products in drinking water standards, and the concentration of each pesti...

Claims

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Application Information

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IPC IPC(8): G01N27/327G01N27/48H01M8/16
CPCG01N27/308G01N27/3277G01N27/3278G01N27/48H01M8/16Y02E60/50
Inventor 刘梅川王宇孙彩琴陆汉星赵国华
Owner TONGJI UNIV
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