Method for detecting microRNA in lung cancer cells based on non-substrate and non-labeled electrocatalytic amplification biosensor

A technology of biosensors and lung cancer cells, applied in biochemical equipment and methods, measurement/inspection of microorganisms, electrochemical variables of materials, etc., can solve the problems of limited detection sensitivity, instability and easy decomposition, etc. The method is simple and low cost , The effect of simplifying the operation procedure

Active Publication Date: 2019-05-07
SHANDONG NORMAL UNIV
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  • 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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  • Method for detecting microRNA in lung cancer cells based on non-substrate and non-labeled electrocatalytic amplification biosensor
  • Method for detecting microRNA in lung cancer cells based on non-substrate and non-labeled electrocatalytic amplification biosensor
  • Method for detecting microRNA in lung cancer cells based on non-substrate and non-labeled electrocatalytic amplification biosensor

Examples

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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, followed by polyadenylation 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. F...

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Abstract

The invention discloses a method for detecting microRNA in lung cancer cells based on a non-substrate and non-labeled electrocatalytic amplification biosensor. The preparation method of the adopted biosensor comprises the following steps of: heating dicyandiamide and ferrous salt to 490-510 DEG C in an inert gas atmosphere to self-assemble to form a precursor, heating the precursor to 890-910 DEGC in the inert gas atmosphere, and performing calcining to obtain iron-containing nitrogen-rich carbon nano-tubes; and modifying the iron-containing nitrogen-rich carbon nano-tubes and gold nano-particles to a glassy carbon electrode to obtain an AuNP/ FeCN/ GCE electrode, and fixing a thiolated capture probe on the AuNP/ FeCN/ GCE electrode through a gold-sulfur bond, wherein the capture probe issingle-stranded DNA capable of hybridizing with target microRNA. In the invention, under the condition of no H2O2, a large amount of thiophen introduced by electrocatalytic reduction greatly enhanceselectrochemical signals.

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