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Application of dual-signal-enhancement paper-based biosensor in miRNA detection

A biosensor and dual-signal technology, applied in the preparation of fluorescence/colorimetric dual-mode sensors, adopts the field of nuclease digestion signal amplification to achieve the effect of improving sensitivity and accurate fluorescence measurement

Inactive Publication Date: 2019-04-16
UNIV OF JINAN
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Nevertheless, high-sensitivity, high-throughput, and specific detection technology is still a challenge

Method used

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  • Application of dual-signal-enhancement paper-based biosensor in miRNA detection
  • Application of dual-signal-enhancement paper-based biosensor in miRNA detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The application of the fluorescent / colorimetric dual-mode paper chip in the detection of let-7a is characterized in that it comprises the following steps:

[0034] (1) Design the hydrophobic wax printing area and hydrophilic working area of ​​the fluorescent layer and colorimetric layer of the paper chip (attached figure 1 ). The paper material of the present invention is chromatographic paper, and the obtained fluorescence / colorimetric dual-mode paper chip pattern is designed with Adobe Illustrator CS4 software. The preparation process of the microfluidic paper chip is as follows: design the microfluidic paper chip on the computer. Hydrophobic wax batch printing patterns, as attached figure 1 As shown, the microfluidic paper chip includes a colorimetric reaction area on the left, a working area in the middle, and a fluorescent reaction area on the right. The diameter of the hydrophilic area is 6 mm, and the channel width is 2 mm;

[0035] (2) Growth of Pt NPs in the ...

Embodiment 2

[0041] The detection steps are the same as in Example 1, except that the detection substance let-7a is replaced with other small molecules such as miRNA or DNA, and the corresponding DNA sequence used in step (3) also needs to be changed.

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Abstract

The invention discloses a preparation method of a low-cost and high-sensitivity fluorescent / colorimetric dual-mode sensor for visual miRNA detection. The sensor is successfully used for miRNA detection. A paper chip is prepared by utilizing a wax printing technology, platinum nanoparticles grow in a working area of a fluorescent layer, the quenching effect of graphite-phase carbon nitride nanosheets on palladium nanoclusters and duplex specific nuclease are utilized to achieve nucleic acid circulation signal amplification, and accordingly accurate measurement of small molecule miRNA is achieved. The prepared paper chip is folded, and visual predetermination can be achieved by dropwise adding a color developing solution in a reaction area; then the paper chip is placed in a fluorescent dishto achieve accurate fluorescent miRNA determination. The operation is simple, convenient and fast, time and effort are saved, and a new reliable method is provided for miRNA detection.

Description

technical field [0001] The invention relates to the technical field of low-cost, high-sensitivity, and visualized analysis and detection of small biological molecules, more specifically, the preparation of a fluorescence / colorimetric dual-mode sensor capable of detecting miRNA. The invention also relates to the use of nuclease cleavage signal amplification technology. Background technique [0002] miRNA is an endogenous small molecule non-coding RNA with a length of about 19-22 mature nucleotides. To date, miRNAs are found in all plants and animals, accounting for approximately 4% of all genomes. At the same time, miRNA has been proven to have an important impact on cell division, growth and apoptosis. Some common diseases, including cancer, cardiovascular and cerebrovascular diseases, and viral infections, are closely related to the abnormal expression of certain specific miRNAs. The exact role and function of each miRNA in a specific organ, tissue or cell remains to be s...

Claims

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Application Information

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IPC IPC(8): C12Q1/6825C12Q1/682
CPCC12Q1/682C12Q1/6825C12Q2525/207C12Q2563/107C12Q2521/345C12Q2565/607C12Q2563/155C12Q2565/629
Inventor 于京华殷雪梅葛慎光
Owner UNIV OF JINAN
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