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Composite membrane modified biosensor and preparation method and application thereof

A biosensor and composite membrane technology, applied in the field of biosensors, can solve the problems of difficult gene sequence detection, inability to conduct online monitoring, and difficulty in analysis and detection, and achieve the effects of improved detection sensitivity, convenient production, and broad application prospects

Inactive Publication Date: 2011-03-02
HUNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the high requirements of the concentration and purity of the sample to be tested by conventional ultraviolet spectrophotometry, it is difficult to analyze and detect gene sequences with low concentration or high turbidity, and online monitoring cannot be carried out
However, due to the limited sensitivity of the existing gene sensing technology, it is not suitable for sub-picomolar (pM, 10 -12 M) gene sequence is also difficult to detect

Method used

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  • Composite membrane modified biosensor and preparation method and application thereof
  • Composite membrane modified biosensor and preparation method and application thereof
  • Composite membrane modified biosensor and preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0053] a kind of like figure 1 The biosensor modified by the composite membrane of the present invention includes a base electrode 1, the base electrode 1 of this embodiment is a glassy carbon electrode, and the sensing end of the base electrode 1 is coated with a 1,5-naphthalene diamine A polymer film 2, the 1,5-naphthalene diamine polymer film 2 is decorated with a multi-walled carbon nanotube-Congo red combination layer 3, and the multi-walled carbon nanotube-Congo red combination layer 3 is adsorbed with nano Gold particle layer 4.

[0054] The biosensor of this embodiment is prepared through the following steps:

[0055] (1) Glassy carbon electrode pretreatment: use conventional methods to pretreat the glassy carbon electrode. Specifically, in this example, the glassy carbon electrode is first suspended on the polishing paper with 0.3 μm and 0.05 μm alumina Polished to the mirror surface, and then sequentially with nitric acid solution (HNO 3 :H 2 O=1:1), absolute eth...

Embodiment 2

[0062] a kind of like image 3 The application of the biosensor modified by the composite membrane of the present invention in the detection of lignin degrading enzymes (this embodiment specifically detects cellobiose dehydrogenase CDH) coding genes specifically includes the following steps:

[0063] (1) Design probe sequences, target strands and PCR amplification primers: select all coding gene sequences corresponding to the CDH produced by Phanerochaete chrysosporium in GenBank, and use DNAMAN software and NCBI BLAST server for sequence comparison, Determine the conserved region of the coding gene sequence corresponding to CDH, and then use Primer Premier 5.0 and Oligo 6.0 software to design capture probes, response probes, target strands, mismatched strands, and PCR primers for the functional genes encoding CDH. All gene sequences were synthesized by Shanghai Sangon Bioengineering Company. The design results are shown in Table 1 below:

[0064] Table 1: Probe sequences, t...

Embodiment 3

[0089] In Example 3 of the present invention, inoculate and cultivate the strain Phanerochaete chrysosporium producing lignin-degrading enzymes, then extract its total DNA, and perform PCR on a shorter fragment containing the target chain gene through a pair of specific amplification primers Amplify, and its product is used as the target chain gene to be tested, and then use the biosensor of Example 1 and the specific method of Example 2 to investigate the specificity and sensitivity of the present invention for the detection of actual cultured products.

[0090] The application of the composite membrane-modified biosensor of this embodiment in the detection of culture products containing lignin-degrading enzyme coding genes specifically includes the following steps:

[0091] (1) Solid-state culture of Phanerochaete chrysosporium: Prepare PDA medium: 20g of potatoes, 2g of glucose, 2g of agar and 100mL of distilled water; fully dissolve the above components, autoclave at 120°C ...

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Abstract

The invention discloses a composite membrane modified biosensor and preparation and application thereof. The biosensor comprises a matrix electrode, the induction end of the matrix electrode is wrapped with an aromatic diamine polymer membrane, a carbon nano tube-Congo red combination layer is modified on the membrane, and a nano gold particle layer is adsorbed on the combination layer. The preparation method comprises the following steps of: forming the polymer membrane at the induction end of the matrix electrode by electric polymerization reaction; and modifying the carbon nano tube-Congo red combination layer on the membrane, and finally electrically depositing nano gold on the combination layer. The biosensor can be applied in detecting target genes; and during detecting, a probe is first designed, then a capture probe is modified, and whether the target genes are contained in hybridization solution to be detected and the concentration of the target genes can be judged by first and second hybridization and enzyme catalytic reaction. The biosensor has the advantages of good electric conductivity, good biocompatibility, high sensitivity, storing anti-interference performance, good selectivity, low manufacturing cost and the like.

Description

technical field [0001] The invention relates to the field of biosensors, in particular to a biosensor modified by a composite membrane, a preparation method and application thereof. Background technique [0002] Biosensor is a subject field with great development potential, and its fast and sensitive detection performance provides indispensable information for many subjects. With the continuous development of nanotechnology and the rapid change of detection methods, the integration of various biosensors based on nanomaterials is an important trend in the development of biosensors. At present, many researchers at home and abroad have conducted a large number of theoretical and experimental studies on the application of carbon nanomaterials and metal nanomaterials in biosensors, and have made breakthroughs, creating a new era for the application of nanomaterials in biosensors. situation. [0003] Carbon nanotubes (CNTs) are good sensing and carrier materials, and their excel...

Claims

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Application Information

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IPC IPC(8): G01N27/26B81C1/00C12Q1/68
Inventor 李贞曾光明汤琳伍梦诗刘灿庞娅章毅黎媛萍雷晓霞
Owner HUNAN UNIV
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