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Mercuric ion or cysteine detecting electrochemical DNA biosensor and manufacturing method and application thereof

A cysteine, electrochemical technology, applied in the field of biosensors, can solve the problems of low detection limit, can not be used to detect a variety of substances, etc., to achieve the effect of low detection limit, wide detection range, strong anti-interference ability

Active Publication Date: 2016-06-15
HUNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of these sensors have good linear range and low detection limit, however most of them are specific to one target object and cannot be used to detect multiple substances

Method used

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  • Mercuric ion or cysteine detecting electrochemical DNA biosensor and manufacturing method and application thereof
  • Mercuric ion or cysteine detecting electrochemical DNA biosensor and manufacturing method and application thereof
  • Mercuric ion or cysteine detecting electrochemical DNA biosensor and manufacturing method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] An electrochemical DNA sensor for detecting mercury ions or cysteine, including a glassy carbon electrode used as a working electrode in a three-electrode system, the surface of the glassy carbon electrode reaction end is modified with a composite film, and the composite film is from the inside to the The composite film is composed of self-doped polyaniline nanofibers, ordered mesoporous carbon and gold nanoparticles arranged in sequence on the outside, and DNA capture probes are self-assembled on the surface of the composite membrane, and the DNA capture probes can be passed through T-Hg 2+ -T structural mismatch forms a hairpin structure.

[0061] The DNA capture probe is the nucleotide sequence of SEQ ID NO.1, specifically:

[0062] 5'-SH-(CH 2 ) 6 - TTCTTTCTTCCCCTTGTTTGTT-3'

[0063] When the above electrochemical DNA sensor is placed in the solution to be tested containing mercury ions, the 3' end "TTGTTTGTT" of the DNA capture probe will fold with the 5' end "T...

Embodiment 2

[0069] A preparation method of the above-mentioned electrochemical DNA sensor for detecting mercury ions or cysteine, comprising the following steps:

[0070] S1. Modified self-doped polyaniline nanofibers (SPAN): 6 μL of SPAN suspension was added dropwise on the surface of the cleaned glassy carbon electrode to obtain a SPAN-modified glassy carbon electrode, namely SPAN / GCE. image 3 Shows the SPAN / GCE in 5.0mM ferricyanide solution containing 0.1MKCl ([Fe(CN) 6 ] 3- / 4- ) of the cyclic voltammograms measured in ) (please confirm).

[0071] The SPAN is synthesized by a one-step chemical oxidation method, and the specific steps are:

[0072] Dissolve 0.22g of cetyltrimethylammonium bromide (CTAB), 0.46g of aniline (AN), and 0.86g of 2-aminobenzenesulfonic acid (ABS) in 80mL of 0.15M hydrochloric acid solution, and then add 40mL of 0.06M ammonium persulfate (APS) solution, put it at 5°C for 24h, and finally filter the resulting dark green precipitate, wash it with deionized wat...

Embodiment 3

[0082] The application of the electrochemical DNA sensor of a kind of embodiment 1 in detecting mercury ion, comprises the following steps:

[0083] (1) Soak the gold nanoparticles / ordered mesoporous carbon / self-doped polyaniline nanofiber-modified glassy carbon electrode reaction ends assembled with DNA capture probes in solutions containing different concentrations of mercury ions for 1 h (mercury ions Concentrations were 1×10 - 6 M, 1×10 -7 M, 1×10 -8 M, 1×10 -9 M, 1×10 -10 M, 1×10 -11 M, 1×10 -12 M, 1×10 -13 M, 1×10 -14 M), take out the glassy carbon electrode and rinse it with TBS buffer solution, then soak it in the solution containing the signal indicator AQDS for 6h, take it out, rinse it and dry it for later use.

[0084] (2) the glassy carbon electrode after step (1) is processed as a working electrode, the saturated calomel electrode is used as a reference electrode, and the platinum electrode is used as a counter electrode to establish a three-electrode sy...

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Abstract

The invention discloses a mercuric ion or cysteine detecting electrochemical DNA biosensor and a manufacturing method and application thereof.The electrochemical DNA biosensor comprises a glassy carbon electrode serving as a working electrode of a three-electrode system; the surface of a reactive end of the glassy carbon electrode is modified by composite membrane composed of auto-doped polyaniline nanofiber, ordered mesoporous carbon and nano-scale gold particles; a DNA capture probe, with the nucleotide sequence SEQ ID NO.1 is assembled on the surface of the composite membrane.The manufacturing method includes steps of modifying auto-doped polyaniline nanofiber and ordered mesoporous carbon, electrodepositing nano-scale gold particles, assembling the DNA capture probe and the like.The mercuric ion or cysteine detecting electrochemical DNA biosensor capable of detecting two different target substances has the advantages of high stability, long service life, high interference resistance capacity, wide detection scope and low limit in detection and is applied to detection of mercuric ion or cysteine.

Description

technical field [0001] The invention relates to the technical field of biosensors, in particular to an electrochemical DNA sensor for detecting mercury ions or cysteine, a preparation method and application thereof. Background technique [0002] As one of the heavy metal pollutants, mercury is highly toxic and has serious hazards to human health and ecosystems. Mercury exists in many forms in the environment, among which mercury ions (Hg 2+ ), as the most common and stable form, is widely distributed in various water bodies. The World Health Organization (WHO) once stipulated that the total content of mercury ions in drinking water should not exceed 0.001mg / L. It can be seen that it is extremely necessary to invent a sensor that is environmentally friendly, has low cost, and can quickly and sensitively detect the concentration of mercury ions in water. [0003] Cysteine ​​(Cys) is a sulfur-containing natural amino acid that plays an important role in biological organisms ...

Claims

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Application Information

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IPC IPC(8): G01N27/26
CPCG01N27/26
Inventor 汤琳谢霞周耀渝曾光明彭博龙蓓青邓垚成
Owner HUNAN UNIV
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