SERS sensor for nucleic acid detection and preparation and multielement detection method thereof

A detection method and sensor technology, applied in the field of functional nanomaterials and biological detection, can solve the problems of lack of joint detection methods, and achieve the effects of good detection sensitivity, no equipment requirements, and simple and effective detection structure.

Inactive Publication Date: 2016-06-08
NANJING UNIV OF POSTS & TELECOMM
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, there are few surface-enhanced Raman scattering sensors for nucleic acid detection, and methods for joint detection of multiple nucleic acid markers in serum are still relatively scarce

Method used

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  • SERS sensor for nucleic acid detection and preparation and multielement detection method thereof
  • SERS sensor for nucleic acid detection and preparation and multielement detection method thereof
  • SERS sensor for nucleic acid detection and preparation and multielement detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1 SERS sensor for detecting microRNA-21 and its preparation and detection methods

[0066] (1) The silver nanorod array SERS substrate is rinsed several times with ultrapure water;

[0067] (2) The probe chain 1 (1uM) was subjected to high-temperature cooling treatment, that is, kept in a constant temperature shaker at 95°C for 5 minutes, and then kept in a refrigerator at 4°C for 20 minutes, to purify the hairpin-type nucleic acid chain;

[0068] (3) Take 20uL probe chain 1 (1uM) and drop it on the SERS substrate, let it stand for 3h at 25°C and 80% humidity, and then wash it with buffer (10mM phosphate, 100mM sodium chloride, pH7.4) Clean; the SERS substrate of the modified probe chain is soaked in buffer (10mM phosphate, 100mM sodium chloride, pH7.4) for 20min to keep the DNA form stable;

[0069] (4) Soak in 1 mM mercaptohexanol solution for 10 min, seal the exposed sites on the surface of the substrate, and gently rinse with buffer solution to prepare a nu...

Embodiment 2

[0074] Example 2 MicroRNA-21 / 486 / 375 ternary serum detection SERS sensor and its preparation and detection method

[0075] (1) The silver nanorod array SERS substrate is rinsed several times with ultrapure water;

[0076] (2) DNA probe chain 1 (1uM) modified with ROX and corresponding to microRNA-21, DNA probe chain 2 (1uM) modified with Cy5 and corresponding to microRNA-486, and DNA probe chain 2 (1uM) modified with FAM and corresponding to microRNA-375 Needle chain 3 (1uM) was subjected to high-temperature cooling treatment respectively, that is, kept in a constant temperature shaker at 95°C for 5 minutes, and then kept in a refrigerator at 4°C for 20 minutes to purify hairpin-type DNA strands;

[0077] (3) Take three kinds of 20uL nucleic acid probe chains (1uM) and drop them on the SERS substrate, let it stand for 3 hours at 25°C and 80% humidity environment, and then wash it with buffer; the SERS substrate of the modified probe chain is in the buffer Soak in solution (10...

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Abstract

The invention discloses an SERS sensor for nucleic acid detection and a preparation and multielement detection method thereof. The sensor is composed of a solid SERS substrate and a nucleic acid probe chain. The nucleic acid probe chain is fixed to the surface of the solid SERS substrate through a gold-sulfur bond, after complementary base pairing with target nucleic acid molecules, sensing detection for nucleic acid is achieved by detecting the SERS signal change of Raman molecules on the probe chain, and exposed loci on the surface of the substrate are sealed through thiol hexyl alcohol. The solid SERS substrate is a silver nanorod array type substrate. The 3' end of the nucleic acid probe chain modifies thiol, the 5' end of the nucleic acid probe chain modifies dye molecules, part of base sequences in the probe chain are complementary to form a hairpin-shaped structure, and part of the base sequences and nucleic acid to be detected can be in complementary pairing. The detection limit of the sensor reaches the fmol level (fM), and joint detection of multiple nucleic acid markers can be achieved. The sensor is easy to prepare, high in detection sensitivity and good in reliability, and has wide application prospects in the fields of early tumor diagnosis and the like.

Description

technical field [0001] The invention belongs to the field of functional nanomaterials and biological detection, and in particular relates to a SERS (Surface Enhanced Raman Scattering) sensor for nucleic acid detection, a preparation method thereof and a multiple detection method. Background technique [0002] In recent years, studies have proved that the uncontrolled expression of miRNA is closely related to the occurrence and development of tumors. miRNA is expressed in specific tissues and developmental stages, with tissue specificity and timing, which determines the important role of miRNA in disease diagnosis. However, the content of these lung cancer markers is very low in the early stage of the disease, and conventional clinical detection techniques are likely to miss the markers due to the limitation of sensitivity; and a single lung cancer marker with good sensitivity and specificity has not been found so far; in addition, The samples for tumor marker detection are g...

Claims

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Application Information

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IPC IPC(8): C12M1/34C12Q1/68
CPCC12Q1/6825C12Q2565/632C12Q2525/207C12Q2525/301
Inventor 宋春元汪联辉杨琰君杨博玥苏邵
Owner NANJING UNIV OF POSTS & TELECOMM
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