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SERS-SPR dual-mode sensor as well as preparation method and application thereof

A sensor and detection method technology, applied in the field of functional nanomaterials and biological detection, can solve problems such as insufficient sensitivity and specificity

Active Publication Date: 2020-10-23
NANJING UNIV OF POSTS & TELECOMM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The present invention proposes a SERS-SPR dual-mode sensor for nucleic acid detection, which can effectively solve the problems of insufficient sensitivity and specificity in most current detection methods

Method used

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  • SERS-SPR dual-mode sensor as well as preparation method and application thereof
  • SERS-SPR dual-mode sensor as well as preparation method and application thereof
  • SERS-SPR dual-mode sensor as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1 Preparation of silver nanohole-nanorod array substrate

[0063] 1. Silver nanohole-nanorod arrays were prepared by nanosphere etching (NSL), reactive ion etching (RIE) and physical evaporation deposition techniques. figure 1 The fabrication process of the silver nanopore-nanorod array is shown. In the first step, 500nm polystyrene (PS) microspheres are assembled on a clean glass or silicon wafer in a monolayer close-packed manner by the air-water interface method to form a microsphere template, a glass or silicon wafer The size is 0.9×2.5em.

[0064] 2. In the second step, use O 2The plasma etches the PS microspheres on the surface of the substrate to reduce the diameter of the microspheres on the surface of the substrate. In the etching process, Trion Technology Phantom III RIE / ICP system was used to operate, the pressure was 40mTorr, the oxygen flow rate was 10sccm, the ICP power was 25W, the RF power was 10W, and the duration was 350s.

[0065] 3. Ph...

Embodiment 2

[0067] Example 2 Characterization of Refractive Index Response Performance of Silver Nanohole-Nanorod Array Substrate

[0068] The silver nanohole-nanorod array substrate (structure such as figure 2 shown) were soaked in different refractive index solvents: methanol (n=1.328), acetone (n=1.359), 1-hexanol (n=1.418), chloroform (n=1.446) and toluene (n=1.496), Afterwards, the polarized and non-polarized transmission spectra of the silver nanohole-nanorod array substrate were tested respectively. Under the irradiation of non-polarized, 0-degree polarized and 90-degree polarized light, the transmission spectrum of the silver nanohole-nanorod array substrate exhibits a refractive index dependence, such as image 3 shown. In addition, both the spectral profile and the peak-to-valley variation of the transmission spectrum exhibit obvious polarization dependence. According to the response of different peaks and valleys to different refractive index solvents, the response of peaks...

Embodiment 3

[0069] Example 3 Preparation of SERS-SPR dual-mode sensor

[0070] 1. Preparation of tetrahedral DNA probes. Such as Figure 5 As shown, tetrahedral DNA is formed by base complementary hybridization self-assembly of four single strands A, B, C, and D. Mix equimolar amounts (1 μM) of four DNA single strands in 300 μL TM buffer (20 mM Tris-HCl, 50 mM MgCl2, pH 8.0), mix them, and anneal (heat to 95 ° C for 5 min, then naturally cool to room temperature) ), forming tetrahedral DNA, the concentration of tetrahedral DNA in the final solution was 1 μM. Tetrahedral DNA formation was characterized by 10% polyacrylamide gel electrophoresis, Figure 6 a is the electrophoretic gel image corresponding to the formation process of tetrahedral DNA. Compared with the combination of one (lane 1), two (lane 2) and three DNA strands (lane 3), four DNA strands are mixed and self-assembled to form a tetrahedron DNA moved the slowest in lane 4, indicating that tetrahedral DNA was formed success...

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Abstract

The invention discloses an SERS-SPR dual-mode sensor for nucleic acid detection. The SERS-SPR dual-mode sensor comprises a detection chip and a DNA probe, wherein the detection chip is a silver nanopore-nanorod array substrate of which the surface is modified with tetrahedral DNA. The invention further discloses a preparation method and a detection method of the SERS-SPR dual-mode sensor. The detection method comprises the following steps of: sequentially mixing the detection chip with a to-be-detected liquid sample and a DNA probe solution; forming a 'detection chip-target DNA-DNA probe' compound through complementary pairing; and then sequentially carrying out a transmission spectrum test and an SERS test. According to the detection method, high-sensitivity and specific dual-mode sensingdetection on nucleic acid in serum is realized through wavelength change of characteristic valleys of a transmission spectrum, an SERS spectrum and characteristic signal intensity values of the SERSspectrum, the detection limits of the SERS sensing mode and the SPR sensing mode reach the sub-femtomole per liter magnitude and the sub-picomole per liter magnitude respectively, and the nucleic acidmarker can be detected in complex environments such as serum.

Description

technical field [0001] The invention belongs to the field of functional nanomaterials and biological detection, and in particular relates to a SERS-SPR dual-mode sensor for nucleic acid detection and its preparation method and application. Background technique [0002] With the continuous development of molecular biology and medical fields, and further research on the mechanism of cancer, people have gradually realized that cancer is a disease caused by changes in genetic material, that is, the corresponding cells secrete cancer markers, including The presence or content of nucleic acid (such as microRNA), protein, hormone, etc. in the blood will reflect the status of cancer. However, in the early stage of lesions, the content of cancer markers is very low. Due to the limitation of sensitivity of conventional clinical detection techniques, it is easy to miss detection during the detection process of markers; in addition, the samples for cancer marker detection are generally ...

Claims

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Application Information

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IPC IPC(8): G01N21/65G01N21/552
CPCG01N21/658G01N21/553
Inventor 宋春元蒋新宇张晶晶汪联辉
Owner NANJING UNIV OF POSTS & TELECOMM
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