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A method for the detection of microRNAs in lung cancer cells based on a substrate-free, non-labeled electrocatalytic amplified biosensor

A biosensor, substrate-free technology, applied in the field of detection of microRNA in lung cancer cells, can solve the problems of instability and easy decomposition, limited detection sensitivity, etc., and achieve the effects of reducing detection cost, simple method and low background signal.

Active Publication Date: 2022-04-15
SHANDONG NORMAL UNIV
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Conventional electrocatalytic amplified electrochemical biosensors usually rely on H 2 o 2 The peroxidase-mediated reaction catalyzed by H 2 o 2 It is unstable and easy to decompose, resulting in limited detection sensitivity

Method used

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  • A method for the detection of microRNAs in lung cancer cells based on a substrate-free, non-labeled electrocatalytic amplified biosensor
  • A method for the detection of microRNAs in lung cancer cells based on a substrate-free, non-labeled electrocatalytic amplified biosensor
  • A method for the detection of microRNAs in lung cancer cells based on a substrate-free, non-labeled electrocatalytic amplified biosensor

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preparation example Construction

[0036] Preparation of iron-containing nitrogen-rich carbon nanotubes (FeCN): Heating dicyandiamide and ferrous salt to 490-510°C in an inert gas atmosphere to form a precursor for self-assembly, and heating the precursor to 890-890°C in an inert gas atmosphere 910°C, iron-containing nitrogen-rich carbon nanotubes were obtained after calcination;

[0037] Iron-containing nitrogen-rich carbon nanotubes and gold nanoparticles (AuNP) were modified to glassy carbon electrodes to obtain AuNP / FeCN / GCE electrodes, and then the thiolated capture probes were immobilized on AuNP / FeCN / GCE electrodes through gold-sulfur bonds, The capture probe is a single-stranded DNA capable of hybridizing with the target microRNA.

[0038] The inert gas mentioned in the present disclosure refers to a gas that can prevent oxidation of oxygen, such as nitrogen, helium, argon, and the like.

[0039] The ferrous salt described in the present disclosure is a compound that can dissolve in water and ionize fe...

Embodiment

[0061] The principle of this embodiment, such as figure 1 Shown:

[0062] The precursor Fe used 2+ -g -C 3 N 4 One-step self-assembly from cheap starting materials including dicyandiamide and iron(II) chloride tetrahydrate ( figure 1 A). One-step synthesis of FeCN can be achieved by treating Fe at 900 °C under argon 2+ Doped graphite Carbon nitride (Fe 2+ -g -C 3 N 4 ) to achieve simple heat treatment. Electrochemical biosensors ( figure 1 B) The construction is as follows: first, FeCN and AuNPs are modified onto the GCE electrode by layer-by-layer self-assembly, and then the thiolated capture probe is immobilized on the GCE electrode that has been modified with FeCN and AuNP through gold-sulfur bonds. The capture probe is then hybridized to the target microRNA, which is then polyadenylated at the 3'-OH end of the microRNA by poly(A) polymerase to form a poly that can further hybridize with the T-rich helper probe to form double-stranded DNA (dsDNA). (A) Sequence. ...

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Abstract

The invention discloses a method for detecting microRNA in lung cancer cells based on a substrate-free and non-labeled electrocatalytic amplified biosensor. Self-assemble at ~510°C to form a precursor, heat the precursor to 890-910°C in an inert gas atmosphere, and obtain iron-containing nitrogen-rich carbon nanotubes after calcination; modify iron-containing nitrogen-rich carbon nanotubes and gold nanoparticles to glass AuNP / FeCN / GCE electrodes were obtained as carbon electrodes, and then thiolated capture probes were immobilized on the AuNP / FeCN / GCE electrodes through gold-sulfur bonds, and the capture probes were single-stranded DNA capable of hybridizing with the target microRNA. The present invention is without H 2 o 2 In the case of , a large amount of thionine introduced by electrocatalytic reduction greatly enhanced the electrochemical signal.

Description

technical field [0001] The invention relates to a method for detecting microRNA in lung cancer cells based on a substrate-free and non-marked electrocatalytic amplification biosensor. Background technique [0002] The statements herein merely provide background information related to the present invention and may not necessarily constitute prior art. [0003] Electrochemical biosensors have attracted increasing attention due to their low cost, good flexibility, high sensitivity, rapidity and portability. The performance of electrochemical biosensors depends on the generation of electrochemical signals and the transfer of electrons on the electrode surface. In order to improve the analytical performance, some electrocatalysts have been introduced, such as noble metals (such as palladium, platinum, and gold), metal oxides (such as MnO 2 , Fe 3 o 4 , and CeO 2 ) and carbon-based nanomaterials for signal amplification. Despite the promising electrocatalytic performance of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12Q1/6825G01N27/327G01N27/48
Inventor 张春阳崔琳王蒙
Owner SHANDONG NORMAL UNIV