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Coupled enhancement SERS (surface enhanced Raman scattering) high-flux biosensing method based on Raman activated nanoparticle mixture assembly for circulating tumor cells

A tumor cell and nanoparticle technology, which is applied in the field of surface-enhanced Raman resonance analysis technology, can solve the problems of complex operation, low separation efficiency, and expensive equipment, and achieves simple operation process, rich spectral information, and good specificity. Effect

Active Publication Date: 2015-04-22
HUNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, there are immunomagnetic bead separation methods for CTC detection, which have low separation efficiency and are easily interfered by white blood cells, resulting in false positive signals; there are reverse transcription-polymerase chain reaction methods, which cannot achieve quantitative analysis of peripheral blood tumor cells ; There is also flow cytometry sorting method, which requires fluorescent labeling, complex operation, time-consuming, and expensive instruments
At present, it is still a great challenge to realize the quantitative analysis of trace CTCs in whole blood with high sensitivity and high specificity.

Method used

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  • Coupled enhancement SERS (surface enhanced Raman scattering) high-flux biosensing method based on Raman activated nanoparticle mixture assembly for circulating tumor cells
  • Coupled enhancement SERS (surface enhanced Raman scattering) high-flux biosensing method based on Raman activated nanoparticle mixture assembly for circulating tumor cells

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Experimental program
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Effect test

Embodiment 1

[0033] Example 1: High-throughput detection of circulating tumor cells

[0034]Two tumor cells (CEM cells and Ramos cells) were selected as model systems, and the nucleic acid sequences used in the experiment were: DNA-1: 5'-HS-TTTTTGGCTTAGGGAAACG (SEQ ID NO.1); DNA-2: 5' - HS-TTTTTCGTTT CCCTAAGCC (SEQ ID NO. 2); aptamer-1: 5'-HS-TTTTTTTTTTATTCTAACTGCTGCGCCGCCGGGAAAA TACTGTACGGTTAGA (SEQ ID NO. 3);

[0035] The specific operation steps are:

[0036] (1) Preparation of Raman dye-labeled gold nanorod probes 1 and 2 that can specifically bind to tumor cells

[0037] Gold nanorod probe 1 specifically binds to membrane proteins of CEM cells, gold nanorod probe 2 specifically binds to membrane proteins of Ramos cells, and the nucleic acid aptamers that can specifically bind to membrane proteins of CEM cells and Ramos cells are labeled as aptamer- 1 and aptamer-2.

[0038] Preparation method of gold nanorod probe 1 modified with raman dye-1 (5,5'-dithiobis(2-nitrobenzoic acid)), a...

Embodiment 2

[0049] Example 2: High-throughput detection of circulating tumor cells in human whole blood

[0050] Two tumor cells (CEM cells and Ramos cells) were selected as model systems, and the nucleic acid sequences used in the experiment were: DNA-1: 5'-HS-TTTTTGGCTTAGGGAAACG (SEQ ID NO.1); DNA-2: 5' - HS-TTTTTCGTTT CCCTAAGCC (SEQ ID NO. 2); aptamer-1: 5'-HS-TTTTTTTTTTATTCTAACTGCTGCGCCGCCGGGAAAA TACTGTACGGTTAGA (SEQ ID NO. 3);

[0051] The specific operation steps are:

[0052] (1) Preparation of Raman dye-labeled gold nanorod probes 1 and 2 that can specifically bind to tumor cells

[0053] Gold nanorod probe 1 specifically binds to membrane proteins of CEM cells, gold nanorod probe 2 specifically binds to membrane proteins of Ramos cells, and the nucleic acid aptamers that can specifically bind to membrane proteins of CEM cells and Ramos cells are labeled as aptamer- 1 and aptamer-2.

[0054] Preparation method of gold nanorod probe 1 modified with raman dye-1 (5,5'-dithiobis(2-...

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Abstract

The invention discloses a coupled enhancement SERS (surface enhanced Raman scattering) high-flux biosensing method based on a Raman activated nanoparticles mixture assembly for circulating tumor cells. The coupled enhancement SERS high-flux biosensing method includes that gold nanorod probes and gold nanoparticles probes in nucleic acid functionalization are prepared, the gold nanorod probes are capable of realizing specific binding with CTCs (circulating tumor cells) and are decorated by Raman dye, the gold nanorod probe decorated with different aptamer are respectively bonded with the corresponding circulating tumor cells, redundant gold nanorod probes are centrifugally removed, the gold nanoparticles probes capable of being hybridized with the gold nanorod probes are added, nanoparticles are agglomerated, and a coupled enhancement effect is realized by the aid of an electromagnetic field. Besides, characteristics of a Raman spectrum are combined, spectrogram information is rich and unique, and high-flux SERS detection for the circulating tumor cells is realized. Besides, the method has the advantages that an operation process is simple and fast, test samples do not need to be separated or enriched, sensitivity is high, specificity is good, and the method can be expected to become a novel method for treatment evaluation and screening of tumor researching chemotherapy drugs.

Description

technical field [0001] The invention belongs to a high-throughput detection method for circulating tumor cells, specifically referring to the specific combination of nucleic acid aptamer (aptamer) and tumor cell membrane protein, functional modification of the surface of nanoparticles, and surface-enhanced Raman resonance (SERS) analysis technology. Background technique [0002] Circulating Tumor Cells (CTCs) refer to tumor cells that enter peripheral blood spontaneously or during diagnostic and therapeutic operations. CTCs with high viability and high metastatic potential can survive in the circulation system and proliferate in a suitable environment, resulting in Tumor recurrence and metastasis. At present, there are immunomagnetic bead separation methods for CTC detection, which have low separation efficiency and are easily interfered by white blood cells, resulting in false positive signals; there are reverse transcription-polymerase chain reaction methods, which cannot...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/65
Inventor 王玉蒋健晖楚霞唐丽娟俞汝勤
Owner HUNAN UNIV