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Cancer detection system based on exosome and method thereof

A cancer detection and exosome technology, applied in the field of exosome-based cancer detection systems, can solve problems such as lack of analysis methods, and achieve the effect of promoting accumulation and convenient operation

Inactive Publication Date: 2018-09-28
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] However, due to the lack of accurate, feasible and easy-to-operate analysis methods, it is still challenging to analyze the small differences in the surface proteins of different exosomes. The development of simple and reliable analysis methods for exosome surface proteins is very important for early diagnosis of tumors

Method used

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  • Cancer detection system based on exosome and method thereof
  • Cancer detection system based on exosome and method thereof
  • Cancer detection system based on exosome and method thereof

Examples

Experimental program
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Embodiment 1

[0082] Incubate exosome samples with fluorescently labeled aptamers, and the selected aptamers are oligonucleotides that can specifically bind to proteins or other small molecular substances screened by the in vitro screening technique SELEX (Systematic Evolution of Ligands by Exponential Enrichment) Fragments, specifically, the fluorescently labeled aptamer is a 40-base single-stranded DNA, and the diameter of the coil in the sample liquid is less than 5 nanometers, while the diameter of the exosome is 30-150 nanometers; it will specifically recognize the CD63 protein Aptamers were applied to exosomes in the culture supernatant of A375 cells (human melanoma cells). Fluorescent groups can be modified at the end of the aptamer by standard means. When the aptamer specifically interacts with the target protein on the surface of the exosome, the exosome is labeled with the fluorescence carried by the aptamer. The exosome sample in this example is the supernatant of the cell cultur...

Embodiment 2

[0087] In this example, using serum samples from patients with cervical cancer, 7 different aptamers were used to detect the abundance of 7 surface proteins (CD63, PTK7, EpCAM, HepG2, HER2, PSA, CA125) of exosomes in serum samples, and Comparison with healthy human serum samples.

[0088] The exosome manipulation method used, as well as the laser, sample chamber, microscope and CCD camera are the same.

[0089] combine Figure 4 As shown, it can be seen that the serum exosomes of this cervical cancer patient highly express CD63 protein, and cancer-related markers PTK7, EpCAM, HepG2, HER2, PSA and CA125, among which CA125 can be used as a traditional cervical cancer marker, and some cervical cancer patients There is high expression of HER2. It is generally believed that tumor markers PTK7 and EpCAM are related to various cancers, HepG2 is mainly specific to liver cancer, and PSA is mainly specific to prostate cancer. However, these tumor markers are not strictly correlated w...

Embodiment 3

[0093] In this embodiment, the micro-nano particles used are non-biological micro-nano particles, specifically fluorescent polystyrene microspheres, the brand is Thermofisher, the diameter is 50 to 200 nanometers, the mass fraction is 0.001%, and it is dissolved in Tween20 containing 0.02%. in aqueous solution. The laser, the sample chamber, the microscope and the CCD camera are all the same as the above-mentioned embodiments 1 and 2.

[0094] combined Figure 5 As shown, all the fluorescent microspheres with different diameters are highly converged at the laser spot, and according to the gray value of the fluorescence measurement and the fluorescence picture, it can be seen that the degree of convergence and the fluorescence intensity increase with the increase of the particle diameter, which is consistent with the work of this embodiment The principle is consistent, that is, large particles tend to aggregate more. This example illustrates that both biological and non-biolo...

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Abstract

The invention relates to a cancer detection system based on an exosome and a method thereof. The cancer detection system comprises a heating unit, a sample chamber unit, and a signal acquisition unit,the exosome combined with a corresponding fluorescent labeled aptamer is loaded in the sample chamber unit, the aptamer is specifically bind to one of target proteins on the surface of the exosome, the heating unit is used for heating the sample chamber unit, the sample chamber unit generates the thermophoresis effect, the exosome is gathered at a low-temperature side in the sample chamber unit,and a fluorescence signal labelled at the exosome in a sample room is amplified; the signal acquisition unit obtains the amplified fluorescence signal to obtain the abundance of the target protein onthe surface of the exosome, various aptamers are respectively combined with different target proteins on the surface of the exosome to obtain the protein spectrum on the surface of the exosome, and canceration detection is carried out. The microscale sample is required for completing the cancer detection, and the application scope is wide.

Description

technical field [0001] The present invention relates to the field of cancer detection, in particular to an exosome-based cancer detection system and method. Background technique [0002] In the prior art, micro-nano particles are detected to measure particle size, shape, concentration, activity, etc., which are widely used in hematology, immunology, molecular biology and other disciplines. In the prior art, the flow particle detection method is often used to detect micro-nano particles, which is a technology for quantitative analysis and sorting of particles in liquid one by one. The Coulter principle used in the detection refers to: suspension When the particles in the electrolyte pass through the small hole with the electrolyte, they replace the same volume of electrolyte, and in the circuit designed by constant current, the resistance between the two electrodes inside and outside the small hole changes instantaneously, generating a potential pulse, the size and number of ...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/533G01N33/558
CPCG01N33/533G01N33/558G01N33/57484G01N2333/4746
Inventor 孙佳姝
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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