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Construction method and application of sers aptamer sensor based on nano-assembly structure

An aptamer sensor and nano-assembly technology, which is applied in the direction of instruments, measuring devices, scientific instruments, etc., can solve the problems of measurement substance interference, inability to guarantee the specificity and selectivity required for detection, complex food matrix components, etc., and achieve simple operation , the effect of improving the sensitivity

Active Publication Date: 2019-03-01
ZHEJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, the composition of the food matrix is ​​complex, and it is easy to interfere with the measured substances. Relying solely on SERS technology cannot guarantee the specificity and selectivity required for detection. Generally, it is necessary to use chemometric methods or establish models for discrimination and identification, especially for OTA. Molecules, the combination of SERS technology and nucleic acid aptamers can just make up for this deficiency

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  • Construction method and application of sers aptamer sensor based on nano-assembly structure
  • Construction method and application of sers aptamer sensor based on nano-assembly structure

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Embodiment Construction

[0046]The present invention will be further described below in conjunction with drawings and embodiments.

[0047] Embodiments of the present invention are as follows:

[0048] 1) Preparation of gold nanorod DNA probe (AuNRs-DNA)

[0049] 1.1) The carboxy-modified first DNA (sequence is 5'-AGTCACAGTGAGACTTTTTTT-CH(6)-SH-3', the base sequence of which is as SEQ ID No.1) is mixed with freshly prepared TCEP solution and heated at room temperature Stand still for 1h, the mol ratio of TCEP and DNA is TCEP:DNA=10:1;

[0050] 1.2) Adjust the pH of 1×TBE buffer solution (containing 0.02% SDS and 500 mM NaCl) to 3.0 with 1 M HCl solution, and then prepare freshly prepared purified gold nanorods (average length: 65.8 nm, The average width is 23.5nm, the radial ratio is 2.8) and the activated DNA solution obtained in step 1.1) is added to the pH-adjusted TBE buffer, mixed and shaken for 10min, and the molar ratio of DNA to gold nanorods is DNA: AuNRs=1000:1, then centrifuged and resus...

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Abstract

The invention discloses a construction method and application of SERS aptasensor based on nano-assembly structure. The method comprises the following steps: respectively modifying surfaces of gold nano-rod and gold nano-particle with two kinds of DNA complementary with an ochratoxin A aptamer segment part to form two nano-probes; then assembling an assembly nano-structure under complementary pairing connection of the two nano-probes and the DNA aptamer, and labeling a Raman beacon molecule on the assembly nano-structure; and adding a solution of a substance for detecting, and detecting the concentration of the assembly nano-structure by utilizing the fact that the SERS signal intensity of the assembly nano-structure is influenced by the substance for detecting. By adopting the method, reaction in a liquid environment is uniform, the detection has high sensitivity and high specificity, only one-step reaction is required, and the operation is simple.

Description

technical field [0001] The invention relates to a construction method and application of a SERS aptamer sensor based on a nano-assembly structure, especially for the application in the detection of ochratoxin A. Background technique [0002] Ochratoxin A (OTA) can cause serious harm to human health through contaminated agricultural products and food, and can cause cancer and even death. Worldwide, the European Union has the strictest limit requirements for mycotoxins, and the limit standard range of OTA in agricultural products and foods is generally between 0.3-50 μg / kg (including infant food). Considering that the sample is generally diluted during extraction, this requires that the developed new detection method must have extremely high sensitivity. At the same time, the composition of the food matrix is ​​complex, which is easy to interfere with the measured substances, so the selectivity or specificity of the method is particularly important. In addition, the detectio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/532G01N21/65
CPCG01N21/658G01N33/532
Inventor 徐霞徐程楠应义斌
Owner ZHEJIANG UNIV