miRNA detection chip, manufacturing method and application thereof

A detection chip and RNA probe technology, applied in the field of miRNA detection chips, can solve the problems of cumbersome operation, easy pollution cost, difficult gene analysis, etc.

Inactive Publication Date: 2015-01-21
SHENZHEN INT TRAVEL HEALTHCARE CENT +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, traditional gene chip technology often requires fluorescent, radioactive, enzyme and other labels or PCR to achieve signal amplification. The operation is too cumbersome, prone to pollution and high cost, and it is difficult to adapt to large-scale gene analysis.

Method used

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  • miRNA detection chip, manufacturing method and application thereof
  • miRNA detection chip, manufacturing method and application thereof
  • miRNA detection chip, manufacturing method and application thereof

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Embodiment Construction

[0024] In the following, the miRNA detection chip, its preparation method and application will be further described in detail mainly in conjunction with the accompanying drawings and specific examples.

[0025] Such as figure 1 As shown, the miRNA detection chip 100 according to one embodiment includes a substrate 110 , a chromium film 120 , a gold film 130 , a self-assembled monolayer 140 and RNA probes 150 .

[0026] The base 110 is a flat glass substrate. The chromium film 120 is disposed on one surface of the substrate 110 with a thickness of 1-2 nm. The gold film 130 is provided on the chromium film 120 with a thickness of 50 nm.

[0027] One end of the self-assembled monolayer 140 formed by 12-mercaptododecanoic acid is fixed on the surface of the gold film 120 through a mercapto group, and the other end is a carboxyl end.

[0028] In this embodiment, the RNA probe 150 includes a specific sequence designed according to the conserved region sequence of respiratory viru...

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Abstract

The invention relates to an miRNA detection chip, which comprises a substrate, a chromium film arranged on the substrate, a gold film located on the chromium film, 12- sulfydryl lauric acid self-assembled monolayer introduced by a self-assembly way on the gold film and an RNA probe combined with the self- assembly monolayer. The specific RNA sequence of the miRNA detection chip can hybridize with cDNA formed by miRNA reverse transcriptase to form a hybridization sequence. When the cDNA formed by reverse transcriptase of the miRNA to be tested can hybridize with the specific RNA sequence, a RNase H is used for enzymatic hydrolysis of the specific RNA sequence in the RNA-cDNA hybrid, and the released cDNA can repeatedly combine with specific RNA sequence on the chip surface, and then hydrolysis is carried out until the RAN probe on the chip surface is completely hydrolyzed, thus forming an amplification detection based on non-PCR. The amplification detection has simple operation, and can realize high throughput and rapid detection of miRNA In addition, the invention also relates to a manufacturing method and application of the miRNA detection chip.

Description

technical field [0001] The invention relates to the field of biochemical detection, in particular to a miRNA detection chip, its production method and application. Background technique [0002] MicroRNA (microRNA, also known as miRNA) is a single-stranded small molecule RNA expressed endogenously in the body, located in the non-coding region of the genome, and has high conservation, timing and tissue specificity. miRNAs can prevent protein expression through imperfectly matched binding to target mRNAs. A single miRNA can regulate thousands of target genes. Studies have shown that miRNA can be produced by viruses. For example, influenza virus expresses 8 specific miRNAs (miRNA-1254, miRNA-1272, miRNA-17-5p, miRNA-17-3p, miRNA-106B, miRNA- 106B, miRNA-124-a and miRNA-124). Research also shows that most miRNAs in serum are wrapped by vesicles, which are resistant to RNases and prevent degradation by nucleases. They are relatively stable and can be stable for 4 hours at room ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C40B40/06C40B50/06
CPCC12Q1/6837C12Q2521/107C12Q2521/327C12Q2525/207C12Q2565/501
Inventor 何建安顾大勇姚旌羽刘春晓史蕾赵纯中徐云庆
Owner SHENZHEN INT TRAVEL HEALTHCARE CENT
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