Construction method and application of SERS aptasensor 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: 2017-10-24
ZHEJIANG UNIV
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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. Gen...

Method used

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  • Construction method and application of SERS aptasensor based on nano-assembly structure
  • Construction method and application of SERS aptasensor based on nano-assembly structure

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Embodiment Construction

[0046] The present invention will be further described below with reference to the drawings and embodiments.

[0047] The embodiment of the present invention is as follows:

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

[0049] 1.1) The first DNA modified by carboxyl group (sequence is 5'-AGTCACAGTGAGACTTTTTTT-CH(6)-SH-3', the base sequence is as SEQ ID No. 1) is mixed with freshly prepared TCEP solution and kept at room temperature Let stand for 1 hour, the molar ratio of TCEP to DNA is TCEP:DNA=10:1;

[0050] 1.2) Adjust the pH of 1×TBE buffer (containing 0.02% SDS and 500mM NaCl) to 3.0 with 1M HCl solution, and then freshly prepare and purified gold nanorods (with an average length of 65.8nm, 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 10 minutes. The molar ratio of DNA to gold nanorods is DNA: AuNRs=1000:1, then centrifuged and resuspended...

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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-assembled structure, in particular to 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 EU has the most stringent requirements for mycotoxins limits. The limits of OTA in agricultural products and foods generally range from 0.3-50μg / kg (including infant food). Considering that the sample extraction process is generally diluted, which requires that the new detection method developed must have extremely high sensitivity. At the same time, the food matrix components are complex and easily interfere with the measured substances, so the selectivity or specificity of the method is particularly important. In addition, the detection method must be simple and practical to ...

Claims

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Application Information

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IPC IPC(8): G01N33/532G01N21/65
CPCG01N21/658G01N33/532
Inventor 徐霞徐程楠应义斌
Owner ZHEJIANG UNIV
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