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

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Problems solved by technology

Currently, there are few surface-enhanced Raman scattering sensors for nucleic acid detection, and

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
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Example Embodiment

[0065] Example 1 SERS sensor for unit detection of microRNA-21 and its preparation and detection method

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

[0067] (2) The probe strand 1 (1uM) is first 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 hairpin nucleic acid strands;

[0068] (3) Take 20uL probe strand 1 (1uM) and drop it on the SERS substrate, let it stand for 3h in an environment of 25℃ and 80% humidity, and then rinse with buffer (10mM phosphate, 100mM sodium chloride, pH7.4) Clean; The SERS substrate of the modified probe chain is soaked in a buffer (10mM phosphate, 100mM sodium chloride, pH7.4) for 20 minutes to keep the DNA morphology stable;

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

Example Embodiment

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

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

[0076] (2) The DNA probe strand 1 (1uM) corresponding to microRNA-21 with modified ROX, the DNA probe strand 2 (1uM) with modified Cy5 corresponding to microRNA-486, and the DNA probe with modified FAM corresponding to microRNA-375 Needle strand 3 (1uM) is separately 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 hairpin DNA strands;

[0077] (3) Take three kinds of 20uL nucleic acid probe strands (1uM) and drop them on the SERS substrate respectively, and let them stand for 3 hours in an environment of 25°C and 80% humidity, and then rinse them with buffer; the SERS substrate of the modified probe strands is in the buffer Soak in th...

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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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