Nucleic acid nano-gold biosensor for detecting mercury ions and kit

A biosensor and nucleic acid nanotechnology, applied in the field of nucleic acid nanogold biosensors, can solve the problems of expensive instruments and complicated operations, and achieve the effects of rapid detection, simple preparation, and high sensitivity

Active Publication Date: 2013-09-11
GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These two methods have the characteristics of high sensitivity, less sample consumption, fast analysis speed, and can realize multi-component analysis. However, the instruments are relatively expensive and the operation is complicated, so it is difficult to be applied in some places lacking equipment resources.

Method used

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  • Nucleic acid nano-gold biosensor for detecting mercury ions and kit
  • Nucleic acid nano-gold biosensor for detecting mercury ions and kit
  • Nucleic acid nano-gold biosensor for detecting mercury ions and kit

Examples

Experimental program
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Effect test

Embodiment 1

[0014] Embodiment 1 Preparation of nucleic acid nano-gold biosensor and kit of the present invention

[0015] 1. Design of four nucleic acid sequences

[0016] In the 1990s, Thomas Carell discovered that the thymine base (T) in nucleic acid can specifically form T-Hg with mercury ions 2+ -T complex, the thermal stability of the complex is higher than that of the A-T base pair. The difference in melting temperature between the double strands formed by the two can reach up to 7°C.

[0017] According to this principle, the present invention has synthesized the following four nucleotide sequences through experimental screening:

[0018] The first segment is an oligonucleotide sequence specifically binding to mercury ions, which contains 64 bases, as shown in SEQ ID NO:1. It includes a segment in Hg 2+ A sequence that, when present, folds into a hairpin conformation, a nicking enzyme (eg, Nt.BbvCI) recognition sequence, and a random sequence.

[0019] SEQ ID NO:1

[0020] 5'-...

Embodiment 2

[0053] Embodiment 2: detection experiment of nucleic acid nano gold biosensor

[0054] The nucleic acid nano-gold biosensor prepared in Example 1 was used to conduct the following experiments to verify its detection effect.

[0055] 1. Prepare mercury ion standard solution gradients with concentrations of 2μM, 1μM, 0.5μM, 0.1μM, 50nM, 10nM, and 5nM, and store at room temperature.

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Abstract

The invention relates to a nucleic acid nano-gold biosensor for detecting mercury ions and a corresponding kit. The nucleic acid nano-gold biosensor comprises a sample pad, a gold-labeled pad, a nitrocellulose membrane and a water-absorbing pad, wherein the nitrocellulose membrane is provided with a detection line and a quality control line; colloidal gold is sprayed on the gold-labeled pad; the detection line is marked on the nitrocellulose membrane after streptavidin and a biotin-labeled SEQ ID NO:3 sequence react; the quality control line is marked on the nitrocellulose membrane after streptavidin and a biotin-labeled SEQ ID NO:4 sequence react; colloidal gold is bonded to an SEQ ID NO:2 sequence, and 5'-terminal of the sequence is modified by sulfydryl. According to the nucleic acid nano-gold biosensor and the corresponding kit, the mercury ions are detected rapidly, simply, conveniently and directly, the defect that a large instrument is required in the conventional detection method is overcome, and the detection sensitivity can be guaranteed.

Description

technical field [0001] The invention relates to the field of metal ion detection, in particular to a nucleic acid nano-gold biosensor for detecting mercury ions and a corresponding kit. Background technique [0002] At present, the traditional methods for detecting mercury ions at home and abroad mainly include spectroscopic analysis, mass spectrometry and chromatography. Spectroscopic analysis refers to the analytical methods that use the principles and experimental methods of spectroscopy to determine the structure and chemical composition of substances, including visible spectrophotometry, atomic absorption spectroscopy, atomic fluorescence spectroscopy, atomic emission spectroscopy, etc. Visible spectrophotometry analysis of mercury ions has low sensitivity and low selectivity, and requires the use of highly toxic substances and volatile organic solvents, which are potentially harmful to experimenters. The experimental conditions of flame atomic absorption spectrometry ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/48C12Q1/34
Inventor 曾令文刘杰陈凌波邢雪荣
Owner GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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