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Preparation method and application of SERS biosensor for detecting tumor marker miRNA-141

A biosensor, tumor marker technology, applied in the field of detection of tumor marker miRNA-141, can solve the problem of undisclosed SERS biosensor and the like

Active Publication Date: 2020-06-19
合肥龙之韵医药技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are no relevant research reports on the preparation method of SERS biosensors for the detection of tumor marker miRNA-141 at home and abroad.

Method used

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  • Preparation method and application of SERS biosensor for detecting tumor marker miRNA-141
  • Preparation method and application of SERS biosensor for detecting tumor marker miRNA-141
  • Preparation method and application of SERS biosensor for detecting tumor marker miRNA-141

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Embodiment 1

[0041] A method for preparing a SERS biosensor for detecting tumor marker miRNA-141, comprising the following steps:

[0042] (1) Preparation of capture unit

[0043] a. Preparation of Fe 3 o 4 Magnetic nanoparticles solution: 0.8 g FeCl 3 ·6H 2 O and 0.4 g FeCl 2 4H 2 Dissolve O in 70 mL of water, pass nitrogen to remove oxygen, stir and mix evenly, heat up to 90 °C, slowly add 20-25wt% ammonia water dropwise, adjust pH = 9-10, continue heating and stirring for 1-2 h, stop heating, and Under the protection of nitrogen, stir and reflux, cool to room temperature, wash with water until neutral, and dilute to 50 mL with ethanol to obtain Fe 3 o 4 magnetic nanoparticle solution;

[0044] b. Preparation of aminated Fe 3 o 4 Magnetic nanoparticles solution: 45 mL Fe 3 o 4 Sonicate the magnetic nanoparticle solution for 0.7 h, add 0.25 mL of 3-aminopropyltriethoxysilane (APTES), stir for 4-5 h, add 1.5 mL of 0.3 mol / L nitric acid solution, continue stirring for 3-4 h, was...

Embodiment 2

[0058] With above-mentioned embodiment 1, its difference is:

[0059] (1) Preparation of capture unit

[0060] a. Preparation of Fe 3 o 4 Magnetic nanoparticles solution: 0.7 g FeCl 3 ·6H 2 O and 0.2 g FeCl 2 4H 2 O dissolved in 50 mL of water;

[0061] b. Preparation of aminated Fe 3 o 4 Magnetic nanoparticles solution: 40 mL Fe 3 o 4 Sonicate the magnetic nanoparticle solution for 0.5 h, add 0.2 mL of 3-aminopropyltriethoxysilane, stir for 4 h, add 1 mL of 0.5 mol / L nitric acid solution, and continue stirring for 3 to 4 h;

[0062] c. Preparation of Fe 3 o 4 @Au magnetic nanoparticle solution: 5 mL of aminated Fe 3 o 4 Magnetic nanoparticle solution and 10 mL of 1 wt% HAuCl 4 The solution was mixed at an ultrasonic frequency of 60 kHz, and after stirring for 1 h, 30 mL of 30 mmol / L trisodium citrate solution was slowly added and ultrasonicated for 2 h;

[0063] d. Prepare capture unit solution: Take 50 µL Fe 3 o 4 The @Au magnetic nanoparticle solution was ...

Embodiment 3

[0077] With above-mentioned embodiment 1, its difference is:

[0078] (1) Preparation of capture unit

[0079] a. Preparation of Fe 3 o 4Magnetic nanoparticles solution: 1.0 g FeCl 3 ·6H 2 O and 0.5 g FeCl 2 4H 2 O dissolved in 100 mL of water;

[0080] b. Preparation of aminated Fe 3 o 4 Magnetic nanoparticles solution: 50 mL Fe 3 o 4 Sonicate the magnetic nanoparticle solution for 1 h, add 0.3 mL of 3-aminopropyltriethoxysilane, stir for 5 h, add 2 mL of 0.1 mol / L nitric acid solution, and continue stirring for 3 to 4 h;

[0081] c. Preparation of Fe 3 o 4 @Au magnetic nanoparticle solution: 10 mL of aminated Fe 3 o 4 Magnetic nanoparticle solution and 20 mL of 1 wt% HAuCl 4 The solution was mixed at an ultrasonic frequency of 100 kHz, and after stirring for 1 h, 50 mL of 20 mmol / L trisodium citrate solution was slowly added, and ultrasonicated for 2 to 3 h;

[0082] d. Prepare capture unit solution: Take 100 µL Fe 3 o 4 The @Au magnetic nanoparticle soluti...

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Abstract

The invention discloses a preparation method and an application of a SERS biosensor for detecting a tumor marker miRNA 141. The method comprises the following steps: (1) combining a Fe3O4@Au magneticnanoparticle with a capture probe to obtain a capture unit solution; (2) combining a hollow gold nanosphere with an initiation probe to prepare a signal unit solution; and (3) reacting and combining acapture unit, miRNA 141 and a signal unit, and performing magnetic separation to obtain the SERS biosensor. The application is characterized in that an SERS signal is detected by a Raman spectrometer, SERS signal intensities corresponding to a series of miRNA-141 with different concentrations are detected, and a quantitative relationship between the concentration of the miRNA-141 and the SERS signal intensity is established; and the concentration of miRNA-141 in an unknown sample is detected according to the quantitative relationship. The SERS biosensor has the advantages of high sensitivity,high specificity, simple assembling process and fast detection speed.

Description

technical field [0001] The invention relates to a detection method for detecting tumor marker miRNA-141, in particular to a preparation method and application of a SERS biosensor for detecting tumor marker miRNA-141. Background technique [0002] Cancer, also known as malignant tumor, is essentially a polygenic disease caused by abnormal cell growth and proliferation mechanism, which seriously endangers human health. The International Agency for Research on Cancer under the United Nations World Health Organization predicts that 13.2 million people will die of cancer by 2030, and the number of confirmed cases will reach 21.4 million. Although the mortality rate of cancer is high, it is not incurable. Studies have shown that early cancer is the best time to cure cancer. Therefore, early detection, early diagnosis and early treatment of tumors can effectively prevent and treat cancer. The discovery of tumor markers makes early diagnosis possible. Tumor markers are substances ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68G01N33/574
CPCG01N33/57484
Inventor 卢静郭智勇贾亚茹邵慧丽胡宇芳王邃
Owner 合肥龙之韵医药技术有限公司
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