Magnetic bead electrochemiluminescence gene sensor-based method for detecting mercury ions and application thereof

A luminescent gene and electrochemical technology, applied in the field of environmental pollutant detection, achieves high accuracy, improved sensitivity, and good specificity

Inactive Publication Date: 2010-11-24
SOUTH CHINA NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Compared with traditional methods, these methods have the advantages of simplicity, economy, speed and high sensitivity, but they all have the disadvantage of limited selectivity. In the ecological environment, trace amounts of mercury ions are always accompanied by a large number of other ions. , so the development of highly sensitive and highly selective mercury ion detection methods is of great significance for environmental detection and food safety monitoring

Method used

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  • Magnetic bead electrochemiluminescence gene sensor-based method for detecting mercury ions and application thereof
  • Magnetic bead electrochemiluminescence gene sensor-based method for detecting mercury ions and application thereof
  • Magnetic bead electrochemiluminescence gene sensor-based method for detecting mercury ions and application thereof

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Embodiment 1

[0047] The method of the invention is applied to mercury ion detection.

[0048] (1) Design two DNA probe sequences, as shown in Table 1, synthesized by Shanghai Sangon Bioengineering Technology Co., Ltd.

[0049] Table 1 DNA probe sequence

[0050]

[0051] (2) A. Preparation of magnetic beaded DNA capture probe 1: Take 100 μl carboxy-modified magnetic beads (particle size 2.8 μm, concentration 10 mg / ml) (Affimag PSC from Tianjin Beisile Company), and use 200 μl imidazole buffer (concentration 100mM, pH 7.0) was washed three times; 500 μl of imidazole buffer (concentration 100 mM, pH 7.0, containing 30 mM carbodiimide hydrochloride (EDC)) was added to the washed magnetic beads, shaken at 300 rpm for 30 minutes, Make the carboxyl group react with EDC to activate the carboxyl group; wash the activated magnetic beads three times with 200 μl imidazole buffer (concentration 100 mM, pH 7.0), and then add 100 μL 500 nM DNA capture probe 1 (as shown in Table 1) , shake at 300 rp...

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Abstract

The invention discloses a magnetic bead electrochemiluminescence gene sensor-based method for detecting mercury ions and an application thereof. In the method, two probes are designed, one is a capture probe to be labelled on the surface of magnetic beads and form a magnetic bead probe; and the other probe is labelled with tris(2,2'-bipyridyl)ruthenium(II) to form an electrochemiluminescence probe. As the two probes have T:T mismatch structures, the two probes can not form a matched double helix structure at room temperature in the absence of mercury ions. When mercury ions exist, the mismatched T:T can form a stable T-Hg2 +-T coordination-complex structure with mercury ions according to the specificity of mercury ions so that mercury ions can mediate the two probes to form the double helix structure. Therefore, the magnetic bead probe can capture the electrochemiluminescence probe on the surface in the presence of mercury ions and then mercury ions can be detected through magnetic bead separation and electrochemiluminescence detection. The method of the invention has high specificity and sensitivity and facilitates the qualitative measurement and quantitative measurement of mercury ions.

Description

technical field [0001] The invention belongs to the technical field of environmental pollutant detection, and in particular relates to a method and application for detecting mercury ions based on a magnetic bead electrochemiluminescence gene sensor. Background technique [0002] With the development of modern industry, heavy metal pollution has caused more and more serious harm to human beings and the environment. Due to its unique chemical and physical properties, mercury is widely used in various industrial production processes, such as electronics industry, mining industry, pesticides, machinery, paper industry, etc. In industrial production, mercury is mostly converted into mercury cations and is Emissions into nature, making mercury ions a major environmental pollutant. Inorganic mercury ions in the environment can be converted into methylmercury by organisms under certain conditions. Inorganic mercury mainly affects the kidneys, while methylmercury mainly affects the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N21/76
Inventor 周小明邢达
Owner SOUTH CHINA NORMAL UNIVERSITY
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