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Method for preparing multi-target DNA biosensor using metal ions as markers

A technology of biosensors and metal ions, which is applied in the field of preparation of multi-target DNA biosensors, to achieve the effects of simple detection process, high sensitivity, fast and easy sensitivity

Inactive Publication Date: 2014-02-19
NORTHWEST NORMAL UNIVERSITY
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Problems solved by technology

However, the use of metal ions as markers for probe DNA has not been reported

Method used

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  • Method for preparing multi-target DNA biosensor using metal ions as markers
  • Method for preparing multi-target DNA biosensor using metal ions as markers
  • Method for preparing multi-target DNA biosensor using metal ions as markers

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Embodiment Construction

[0024] Used instrument and reagent in the implementation process of the present invention:

[0025] CHI 660C electrochemical workstation (Shanghai Chenhua Instrument Co., Ltd.) was used to conduct square wave anodic stripping voltammetry and cyclic voltammetry experiments. Electronic balance (Beijing Sartorius Instrument Co., Ltd.) was used to weigh medicines. An ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.) was used to ultrasonically clean the electrodes. Aluminum oxide grinding powder (0.30 , 0.05 , Shanghai Chenhua Instrument Reagent Company) for processing glassy carbon electrodes. Ag / AgCl is the reference electrode, and platinum wire is the counter electrode. Metallothionein (purity greater than 99%, Dalian United Botai Biotechnology Co., Ltd.), 4-(N-maleimidomethyl) cyclohexane-1-carboxylic acid succinimidyl ester (Pierce), Dithiothreitol, hexanethiol (Aladdin), bismuth chloride (Shanghai Reagent General Factory No. 2 Branch Factory), zinc sulfate, c...

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Abstract

The invention provides a method for preparing a multi-target DNA (Deoxyribose Nucleic Acid) biosensor using metal ions as markers. According to the invention, the method that the metal ions are used as the markers to construct the DNA biosensor and metallothionein (MT) is used as a bridge to achieve connection of the metal ions and a DNA detection probe sequence is proposed for the first time. Firstly, Zn<2+>, Cd<2+> and Pb<2+> are respectively marked on detection probes of three virus DNA segments by the metallothionein, then the marked detection probes are crossbred with corresponding DNA capture probes fixed on a gold substrate and target DANs in samples to form a sandwich structure, and finally, the metal ions are dissociated by hydrochloric acid and are detected by an anodic-stripping voltammetric method, so that the purpose of detecting the target DNAs is achieved. The sensor prepared by the method can simultaneously detect various target DNAs and has the characteristics of high sensitivity and low limit of detection.

Description

technical field [0001] The invention relates to a preparation method of a multi-target DNA biosensor marked with metal ions. [0002] Background technique [0003] The bismuth film electrode is sensitive to heavy metal ions (such as Cu + , Zn 2+ 、Cd 2+ , Pb 2+ etc.) with excellent electrochemical response. Similar to mercury electrodes, bismuth ions and heavy metal ions can be reduced to alloys under the action of reduction potential, and the alloys can be re-oxidized to metal ions, showing sharp and symmetrical oxidation signals on the electrode. Compared with mercury electrodes, bismuth is an environmentally friendly material that has been favored by scientists in recent years. Studies have shown that bismuth film electrodes can not only detect different heavy metal ions simultaneously, but also have a low detection limit. However, the use of metal ions as labels for probe DNA has not been reported yet. [0004] Metallothionein (MT) is a class of low molecular weig...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/48
Inventor 刘秀辉郑李纯刘元香汪维维
Owner NORTHWEST NORMAL UNIVERSITY
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