Method and device for detecting carcinoembryonic antigen content

A carcinoembryonic antigen and detection method technology, which is applied in the field of biomedical analysis and detection, can solve the problems of low technical sensitivity and inability to detect early cancer, and achieve the effect of taking into account the separation effect, accurate carcinoembryonic antigen detection results, and reliable detection results

Inactive Publication Date: 2014-07-02
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Aiming at the above defects or improvement needs of the prior art, the present invention provides a carcinoembryonic antigen detection method and device, the purpose of which is to detect lo

Method used

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  • Method and device for detecting carcinoembryonic antigen content
  • Method and device for detecting carcinoembryonic antigen content

Examples

Experimental program
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Effect test

Embodiment 1

[0048] A carcinoembryonic antigen detection method, comprising the following steps:

[0049] (1) Preparation of carcinoembryonic antigen aptamer-fluorescent substance: the concentration was 10 -5 M. 30 μL of sodium thioglycolate-modified semiconductor zinc sulfide coated cadmium selenide core-shell (CdSe / ZnS) quantum dots mixed with a linker, the linker is 1-ethyl-3-(3-dimethylamino Propyl) carbodiimide hydrochloride and N-hydroxysulfosuccinimide molar ratio 5:1 mixture, this mixture is mixed with 50 μ L, 120 μ M carcinoembryonic antigen aptamer buffer solution, the buffer The solution is 0.2M phosphate buffer solution, the pH value is 8.4, so that the molar ratio of fluorescent substance, linker and carcinoembryonic antigen aptamer is 1:1000:20, so that condensation reaction occurs between fluorescent substance and aptamer After reacting for 3 hours, the mixture was centrifuged and ultrafiltered to remove excess carcinoembryonic antigen aptamer to obtain a carcinoembryonic a...

Embodiment 2

[0061] A carcinoembryonic antigen detection method, comprising the following steps:

[0062] (1) Preparation of carcinoembryonic antigen aptamer-fluorescent substance: the concentration was 10 -7 M. 30 μL bovine serum albumin-modified fluorescent nano-gold clusters are mixed with a linker, the linker is an aqueous solution of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride, and The mixture was mixed with 50μL, 6×10 -7 The carcinoembryonic antigen aptamer buffer solution of μ M is mixed evenly, and described buffer solution is the phosphate buffered saline solution of 0.2M, and pH value is 8.0, makes the molar ratio of fluorescent substance, linking agent and carcinoembryonic antigen aptamer be 1: 500:10, so that the condensation reaction between the fluorescent substance and the aptamer occurs. After 2 hours of reaction, the mixture is centrifuged and ultrafiltered to remove excess carcinoembryonic antigen aptamer, and the carcinoembryonic antigen-containing apta...

Embodiment 3

[0074] A carcinoembryonic antigen detection method, comprising the following steps:

[0075] (1) Preparation of carcinoembryonic antigen aptamer-fluorescent substance: the concentration was 10 -6 M. 30 μL of fluorescent nano-silver clusters mixed with linker, the linker is 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysulfosuccinide A 1:1 mixture of amines, the mixture was mixed evenly with 50 μL, 3 μM carcinoembryonic antigen aptamer buffer solution, the buffer solution was a 0.2M phosphate buffer solution with a pH value of 7.4, so that the fluorescent substance, the connecting The molar ratio of the reagent and the CEA aptamer is 1:200:5, so that a condensation reaction occurs between the fluorescent substance and the aptamer. After 4 hours of reaction, the mixture is centrifuged and ultrafiltered to remove excess CEA aptamer. Ligand, the obtained carcinoembryonic antigen-containing aptamer-fluorescent substance was dissolved by adding 100 μL of...

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Abstract

The invention discloses a method and a device for detecting the carcinoembryonic antigen content. The method comprises the following steps of (1) uniformly mixing a fluorescent substance with a connection agent to obtain a first mixture, and then mixing the first mixture with a carcinoembryonic antigen aptamer solution; (2) uniformly mixing a mixed solution with a graphene oxide solution to cause fluorescence quenching so as to obtain a carcinoembryonic antigen aptamer-phosphor/graphene oxide solution; (3) uniformly mixing a sample to be detected and the carcinoembryonic antigen aptamer-phosphor/graphene oxide solution to cause fluorescence recovery; and (4) performing capillary electrophoresis, performing fluorescent detection at a position meeting the requirement that the distance from the position to a positive electrode is 1/2-2/3 of the total length of a capillary, and measuring the concentration of a carcinoembryonic antigen. The device comprises a capillary electrophoresis device and a fluorescence inspection device, wherein the total length of the capillary of the capillary electrophoresis device is 50-100cm, and the inner diameter is 50-100 microns; a detection window is arranged in a position meeting the requirement that the distance from the position to a positive electrode buffering solution tank is 1/2-2/3 of the total length of the capillary; the fluorescence inspection device is arranged at the capillary detection window. The method and the device are high in sensitivity, low in detection limit and high in detection efficiency and can be applied to early screening of cancers.

Description

technical field [0001] The invention belongs to the field of biomedical analysis and detection, and more specifically relates to a method and device for detecting carcinoembryonic antigen content. Background technique [0002] Carcinoembryonic antigen (CEA) is a broad-spectrum tumor marker, which has important reference significance for the diagnosis and screening of colorectal cancer, breast cancer, lung cancer, liver cancer, ovarian cancer and other tumors. It is a good marker for the judgment of curative effect, disease development, detection and prognosis estimation. Therefore, it is of great significance to detect it with high sensitivity and high specificity in clinical medicine. [0003] At present, the methods for detecting the content of carcinoembryonic antigen include immunoassay, such as colorimetric immunoassay, fluorescence immunoassay, electrochemical immunoassay, chemiluminescence immunoassay, electrochemiluminescence immunoassay, etc. Aptamer and fluoresce...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/533
CPCG01N33/57473G01N33/54346G01N2550/00
Inventor 赵元弟周子明方碧云
Owner HUAZHONG UNIV OF SCI & TECH
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