Specific detection method of human breast cancer cells MCF-7 based on surface-enhanced Raman spectroscopy

A surface-enhanced Raman, human breast cancer cell technology, applied in Raman scattering, material excitation analysis, etc., can solve the problems of cell destruction and invasiveness, and achieve strong specificity, high sensitivity, and high Raman signal enhancement ability Effect

Inactive Publication Date: 2012-07-25
NANJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Current technologies for single-cell research mainly include fluorescence spectroscopy, scanning probe microscopy, microfluidic technology, and capillary electrophoresis, but most of them are invasive and can damage cells.

Method used

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  • Specific detection method of human breast cancer cells MCF-7 based on surface-enhanced Raman spectroscopy
  • Specific detection method of human breast cancer cells MCF-7 based on surface-enhanced Raman spectroscopy
  • Specific detection method of human breast cancer cells MCF-7 based on surface-enhanced Raman spectroscopy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Dissolve the aptamer (S2.2, sequence 5'-GCA GTT GAT CCT TTG GAT ACC CTG G-3') in 0.1 mol / L phosphate buffer (PBS, pH 7.4) at a concentration of 300 ng / mL ; the NaAuCl of 1mol / L 4 and 1mol / L AgNO 3 After the solution is mixed evenly at a volume ratio of 1:1, it is added to the nucleic acid aptamer solution, and the Au 3+ 、Ag + The mass ratio of base and base is 1:1:5. Stir the mixture continuously at 4°C for 0.5–2 hours, store it in the dark for 12 hours, and transfer it to a 1cm×1cm quartz cuvette In this method, the Au-Ag-aptamer nanocomposite was obtained by irradiating for 30 minutes under a UV lamp with a wave number of 254 nm; a 0.1 mol / L Rh 6G methanol solution was prepared and added to the Au-Ag-aptamer nanocomposite dispersion In the solution, control the final concentration of Rh 6G to 1 μmol / L, stir at 4 °C for 1 h in the dark, centrifuge at 10,000 rpm for 10 min, and wash the precipitate with 0.1 mol / L PBS solution (pH 7.4) for a total of 3 times , to rem...

Embodiment 2

[0034] Dissolve the aptamer (S2.2, sequence 5'-GCA GTT GAT CCT TTG GAT ACC CTG G-3') in 0.1 mol / L phosphate buffer (PBS, pH 7.4) at a concentration of 300 ng / mL ; the NaAuCl of 1mol / L 4 and 1mol / L AgNO 3 After the solution is mixed uniformly at a volume ratio of 2:1, it is added to the nucleic acid aptamer solution, and the Au 3+ 、Ag + The mass ratio of base and base is 2:1:5, and the mixture is continuously stirred at 4°C for 0.5–2 hours, and stored in a dark place away from light for 12 hours, and then transferred to a 1cm×1cm quartz cuvette In this method, the Au-Ag-aptamer nanocomposite was obtained by irradiating for 30 minutes under a UV lamp with a wave number of 254 nm; a 0.1 mol / L Rh 6G methanol solution was prepared and added to the Au-Ag-aptamer nanocomposite dispersion In the solution, control the final concentration of Rh 6G to 1 μmol / L, stir at 4 °C for 1 h in the dark, centrifuge at 10,000 rpm for 10 min, and wash the precipitate with 0.1 mol / L PBS solution (...

Embodiment 3

[0036] Dissolve the aptamer (S2.2, sequence 5'-GCA GTT GAT CCT TTG GAT ACC CTG G-3') in 0.1 mol / L phosphate buffer (PBS, pH 7.4) at a concentration of 300 ng / mL ; the NaAuCl of 1mol / L 4 and 1mol / L AgNO 3 After the solution is mixed uniformly at a volume ratio of 4:1, it is added to the nucleic acid aptamer solution, and the Au 3+ 、Ag + The mass ratio of base and base is 4:1:5. Stir the mixture continuously at 4°C for 0.5–2 hours, and store it in the dark for 12 hours, and transfer it to a 1cm×1cm quartz cuvette In this method, the Au-Ag-aptamer nanocomposite was obtained by irradiating for 30 minutes under a UV lamp with a wave number of 254 nm; a 0.1 mol / L Rh 6G methanol solution was prepared and added to the Au-Ag-aptamer nanocomposite dispersion In the solution, control the final concentration of Rh 6G to 1 μmol / L, stir at 4 °C for 1 h in the dark, centrifuge at 10,000 rpm for 10 min, and wash the precipitate with 0.1 mol / L PBS solution (pH 7.4) for a total of 3 times ,...

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Abstract

The invention relates to a specific detection method of human breast cancer cells MCF-7 based on surface-enhanced Raman spectroscopy. MCF-7 cells act with a probe compound Rh6G-Au-Ag-aptamer, a characteristic absorption peak of Rh6G appears on the obtained Raman spectrum, and MCF-7 cells are detected by surface-enhanced Raman spectroscopy. The probe compound is prepared by the steps of using nucleic acid apatamer S2.2 as a template, depositing gold and silver bimetal alloy nanoparticles on the bases of the DNA chain of the nucleic acid apatamer S2.2 by using a photocatalysis method, and carrying a Raman signaling molecule Rh6G on the surface of the nucleic acid apatamer S2.2. The method is based on the probe compound which is in target binding with the MCF-7 cells and has high surface-enhanced Raman activity, and carries out specific detection of MCF-7 cells by using the specific target binding capacity of the apatamer with the MCF-7 cells and the enhancement effect of the alloy nanoparticles on the surface of the apatamer on the Raman signal of the Rh6G molecule.

Description

technical field [0001] The invention belongs to the technical field of cell detection methods, and relates to a detection method for human breast cancer cell MCF-7, in particular to a method for specifically detecting human breast cancer cell MCF-7 based on surface-enhanced Raman spectroscopy, which is a sensitivity A method for detecting human breast cancer cell MCF-7 with high and specificity. Background technique [0002] Cancer is an important cause of human death. According to the statistics of the World Health Organization, about 7 million people die of various cancer diseases every year. Among women, breast cancer is the cancer with the highest incidence rate, and the incidence rate of breast cancer has shown an obvious upward trend in recent years. In 2008, the incidence rate of breast cancer in the world was as high as 41.4%. Early diagnosis and treatment of breast cancer are of great significance to reduce breast cancer mortality and improve the 5-year survival ra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/65
Inventor 吴萍蔡称心张卉
Owner NANJING NORMAL UNIVERSITY
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