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Tumour-associated protein detection method based on monomolecular counting

A tumor-related and detection method technology, applied in the field of detection of tumor-related proteins based on single-molecule counting, can solve the problems of low sensitivity, high detection limit, false positive reaction, etc., and achieve high detection sensitivity, easy operation, and sample use Small amount of effect

Active Publication Date: 2013-06-12
INST OF CHEM CHINESE ACAD OF SCI
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Problems solved by technology

[0003] The protein detection method represented by enzyme-linked immunosorbent assay (ELISA) is one of the most common protein detection technologies on the market at present. Ultra-low concentration detection of protein; at the same time, the use of biological enzymes to amplify the signal may cause false positive reactions and affect the authenticity of the detection

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  • Tumour-associated protein detection method based on monomolecular counting
  • Tumour-associated protein detection method based on monomolecular counting
  • Tumour-associated protein detection method based on monomolecular counting

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Embodiment 1

[0031] 1. Total Internal Reflection Microscope

[0032] Such as figure 1 Shown, the total internal reflection microscope among the present invention comprises following optical parts: laser device 1, excitation color filter 2, spectroscope 3, objective lens 4, glass plate 5, reflecting mirror 6, emission color filter 7 and detector CCD 8. The objective lens is a 100X special objective lens for total internal reflection with a numerical aperture of 1.45, and lens oil needs to be added when using it; the sample stage is used to carry and capture the protein glass sheet (refractive index 1.52) to be tested, and the detector is a high-sensitivity CCD. The working process of the optical path is as follows: the laser is used as the excitation light source to emit laser light, the laser light passes through the emission filter and reaches the beam splitter, then is reflected into the objective lens, and finally reaches the sample; the sample emits fluorescence after being excited, t...

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Abstract

The invention discloses a tumour-associated protein detection method based on monomolecular counting. The tumour-associated protein detection method provided by the invention includes the following steps: 1) the tumour-associated protein molecule to be detected is captured onto a glass sheet by specificity interaction of ligand and receptor or specificity interaction of antibody and antigen, thenfluorescent labelled antibody directing to the tumour-associated protein molecule to be detected is added on the glass sheet, so as to obtain the glass sheet to be detected; 2) the glass sheet to be detected obtained by the step 1) is placed on the objective lens of a total internal reflection microscope to carry out detection of tumour-associated protein. The invention belongs to protein detection method, has the advantages of convenient operation, high detection sensibility, less sample usage and no enzyme amplification compared with similar technology and can detect low concentration protein molecule.

Description

technical field [0001] The invention relates to a method for detecting tumor-associated proteins based on single-molecule counting. Background technique [0002] Cancer is the leading cause of death among residents in my country. Early detection and early intervention of cancer can effectively improve the cure rate of cancer. In the early stages of cancer, tumor markers represented by tumor-associated proteins can be used as important indicators for cancer diagnosis. Therefore, the development of ultra-sensitive and ultra-low detection limit protein analysis and detection methods is an inevitable requirement for the ultra-sensitive detection of tumor markers. It is of great significance to the early detection of tumors and early diagnosis of other important diseases, and it is also a current hot research direction in the field of biochemical analysis. [0003] The protein detection method represented by enzyme-linked immunosorbent assay (ELISA) is one of the most common p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/574G01N33/577G01N33/52
Inventor 方晓红张伟周翠松
Owner INST OF CHEM CHINESE ACAD OF SCI
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