Micro-fluidic filter chip, AuNPs-based nucleic acid triple detection kit and AuNPs-based nucleic acid triple detection method

A technology of microfluidics and kits, applied in the direction of supporting/immobilizing microorganisms, sterilization methods, chemical instruments and methods, etc., can solve the problem of low detection accuracy of colorimetry, and achieve simple assembly and practicability Extensive, easy-to-use effects

Pending Publication Date: 2021-06-22
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to solve the problem that the detection accuracy of the existing AuNPs colorimetric method is not high, and to provide a microfluidic chip, kit and method for miRNA triple detection based on AuNPs

Method used

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  • Micro-fluidic filter chip, AuNPs-based nucleic acid triple detection kit and AuNPs-based nucleic acid triple detection method
  • Micro-fluidic filter chip, AuNPs-based nucleic acid triple detection kit and AuNPs-based nucleic acid triple detection method
  • Micro-fluidic filter chip, AuNPs-based nucleic acid triple detection kit and AuNPs-based nucleic acid triple detection method

Examples

Experimental program
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Effect test

Embodiment 1

[0042] Embodiment 1: the first microfluidic filter chip provided by the present invention, such as figure 1 As shown, the structure of the chip includes a suction layer 1 , a sample application layer 2 , a filter layer 3 , a detection layer 4 , and a base layer 5 sequentially from top to bottom.

[0043] Such as figure 2 As shown, the suction layer 1 is provided with a suction port 11, a first micro-channel 12 connected to the suction port 11, and a through hole for connecting the lower sample loading layer. Wherein, the suction port 11 is used for connecting a micropump.

[0044] Such as image 3 As shown, a first microcavity 13 and a fourth microcavity 14 are provided on the sample application layer 2 . Wherein, the first microcavity 13 corresponds to the through hole reserved on the pumping layer 1 , and is used for pouring the mixture into the first microcavity 13 from above the pumping layer 1 . The first micro-channel 12 is connected with the fourth micro-cavity 14,...

Embodiment 2

[0050] Example 2 The second microfluidic filter chip provided by the present invention, such as Figure 7 As shown, its main structure is the same as that of the microfluidic filter chip in Example 1, the only difference is that the chip in this example does not contain a poly-lysine substrate, and a Valve control layer 6, such as Figure 8 As shown, on the valve control layer 6, a water injection port 19 and a valve 21 are provided to connect the valve and the flow channel 20 of the water injection port.

[0051] The working principle and usage method of the microfluidic filter chip of this embodiment are as follows: after the chip is assembled, the mixture solution is poured into the first microcavity 13 from above the suction layer 1, and at the suction port 11 of the suction layer 1 The negative pressure suction force provided by the micropump is used as the driving force, which acts on the first microchannel 12, the fourth microcavity 14, the third microcavity 18, and th...

Embodiment 3

[0053] Embodiment 3 The third embodiment provided by the present invention is: a nucleic acid triple detection kit based on AuNPs, which includes: a microfluidic filter chip as provided in Embodiment 1, a Cy3 fluorescent group modified DNA probe solution, AuNPs solution; miRNA standard solution.

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Abstract

The invention belongs to the technical field of nucleic acid detection, and relates to a micro-fluidic chip, a kit and a method for nucleic acid detection. The micro-fluidic filter chip sequentially comprises a sample adding layer, a filter layer and a detection layer from top to bottom, wherein the sample adding layer is provided with a first microcavity; a filter membrane is arranged at a position, corresponding to the first microcavity, on the filter layer; a second microcavity is formed in the position, corresponding to the filtering membrane, of the detection layer; and the mixture entering the first microcavity enters the second microcavity through the filtering membrane under the action of driving force, so that solid-liquid separation is realized. According to the invention, an AuNPs solution after colorimetric reaction is filtered and separated by using the chip, along with the increase of miRNA concentration, formed double strands are increased, DNA probes adsorbed on the surface of AuNPs are reduced, and the Raman spectrum intensity of fluorophores which stay on a filter membrane and are combined with the DNA probes is reduced; the fluorescence intensity of free double chains in the solution is higher and higher, and triple verification is performed in combination with a colorimetric result, so that the detection accuracy is improved.

Description

technical field [0001] The invention belongs to the technical field of nucleic acid detection, and relates to a microfluidic chip, a kit and a method for nucleic acid detection. Background technique [0002] In recent years, researchers have successively found that miRNA is abnormally expressed in the early stage of cancer, and it has received more and more attention as a new type of tumor marker. Traditional miRNA detection methods such as Northern blot, PCR and microarray. These methods require transcription and amplification, and the detection steps are cumbersome, time-consuming and labor-intensive. [0003] For absolute or relative quantitative detection of miRNAs, many novel sensing methods have been developed in the prior art by using colorimetry, fluorescence, SERS, electrochemistry, and surface plasmon resonance as detection platforms. Among them, the colorimetric method is simple, low-cost, easy to operate, and can monitor signal changes intuitively, so it has at...

Claims

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Application Information

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IPC IPC(8): C12M1/00C12M1/36C12M1/34C12M1/12B01L3/00
CPCC12Q1/6816B01L3/5027B01L3/502753B01L2200/10B01L2300/0887C12Q2525/207C12Q2563/107C12Q2565/629C12Q2565/632C12Q2563/137
Inventor 韩琳高亚坤张宇
Owner SHANDONG UNIV
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