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SERS biosensor and application thereof in preparation of detection system for detecting myocardial infarction miRNA

A biosensor and system technology, applied in the field of bioanalysis and detection, can solve the problem of low sensitivity

Pending Publication Date: 2021-11-09
ANHUI UNIVERSITY OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the very small cross-section of Raman spectroscopy, its sensitivity is very low

Method used

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  • SERS biosensor and application thereof in preparation of detection system for detecting myocardial infarction miRNA
  • SERS biosensor and application thereof in preparation of detection system for detecting myocardial infarction miRNA
  • SERS biosensor and application thereof in preparation of detection system for detecting myocardial infarction miRNA

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] The hybrid chain amplification strategy was confirmed on a 1.5% agarose gel. Firstly, the two hairpin rings S2-HP1 and S2-HP2 were individually modified on AuNPs of about 16nm, and then, different concentrations of S1-HP2 (customized, the same as the product chain after enzyme digestion) and 10mM MgAc 2 The mixture was incubated at 37°C for 3 hours to form a hybrid chain amplification reaction. Electrophoresis was performed at 8 V / cm 3 in 0.5×TBE buffer. The final electrophoresis results are captured by a smartphone, such as figure 2 .

[0055] SERS sensor preparation: first take 500μL, 0.2mg / mL Fe 3 o 4 , add EDC and NHS and shake overnight (10mMEDC 5μL, 1mM NHS 5μL), the Fe 3 o 4 activation. Then the hairpin probe S1-HP and the above-mentioned activated Fe 3 o 4 Add it to 1 mL of PBS buffer with a pH of 7.4, incubate with shaking at room temperature for more than 10 h, and use H with the aid of a magnet. 2 O was washed once and redissolved in PBS buffer. T...

Embodiment 2

[0061] In Example 2 on the basis of Example 1, the detection target miRNA-499 was replaced with the detection target miRNA-328, and the steps and methods were the same as in Example 1. Such as Figure 6is the relationship graph and linear relationship between the concentration of miRNA-328 and Raman intensity, adding a series of miRNA-328 with a concentration ranging from 1fM to 1nM can significantly enhance the Raman intensity, which makes the relationship between Raman intensity and miRNA-328 concentration There is a linear relationship between the numbers. The effect is the same as miRNA-499.

Embodiment 3

[0063] In Example 3 on the basis of Example 1, the detection target miRNA-499 is replaced by the detection target miRNA-208, such as Figure 7 is the relationship graph and linear relationship between the concentration of miRNA-208 and Raman intensity, adding a series of miRNA-208 with a concentration ranging from 1fM to 1nM can significantly enhance the Raman intensity, which makes the relationship between Raman intensity and miRNA-208 concentration There is a linear relationship between the numbers. The effect is the same as miRNA-499.

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Abstract

The invention relates to an SERS biosensor; hairpin probe DNA is modified on the surface of Fe3O4; MCH is added for incubation to prevent non-specific binding; miRNA, auxiliary chain DA and auxiliary chain DB are added to generate an enzyme digestion product; hairpin DNA 1 and hairpin DNA 2 are modified on the surface of AuNP in advance; the dispersed AuNP modified by the hairpin DNA 1 and the hairpin DNA 2 is added into the incubated Fe3O4 to carry out a hybrid chain amplification reaction; reaction is carried out overnight; and centrifugal washing is carried out. According to the invention, in a nano-assembly SERS detection system, for unreacted gold nano-cluster elements and aggregates generated by non-triggering, Fe3O4 is adopted as a magnetic separation substrate; an assembly triggered by hybrid chain amplification is rapidly separated out through surface functionalization and exclusion of non-specific adsorption, so that the aggregates only containing gold nanoparticles on the Fe3O4 surface are obtained; a very high SERS signal is achieved; and myocardial infarction miRNA detection is achieved.

Description

technical field [0001] The invention relates to the field of biological analysis and detection, in particular to a SERS biosensor and its application in the preparation of a detection system for detecting myocardial infarction miRNA Background technique [0002] MicroRNA (miRNA) is a class of single-stranded RNA molecules of 18–25 bases. Current studies have confirmed that there are disease-related miRNAs in the blood circulation system, these circulating miRNAs are not constant, and usually change with the development of the disease, in different life activities of different organs, tissues, and cell types , the type and expression of miRNA are also very different. More and more studies have shown that with the development of major diseases such as cancer and myocardial infarction, the expression of circulating miRNAs will be dysregulated. Therefore, miRNA in blood has been proved to be an important marker for disease diagnosis and prevention. Analysis of circulating miR...

Claims

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Application Information

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IPC IPC(8): C12Q1/682C12Q1/6883
CPCC12Q1/682C12Q1/6883C12Q2600/158C12Q2600/178C12Q2525/301C12Q2521/345C12Q2565/632C12Q2563/143C12Q2563/149C12Q2563/137C12Q2563/155
Inventor 孙玉蝶易洋刘佳旭方腊张奎
Owner ANHUI UNIVERSITY OF TECHNOLOGY
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