Preparation and applications of electrochemical immunosensor used for detecting circulating tumor cells

A tumor cell and sensor technology, applied in the field of electrochemical cell immunosensor preparation, can solve the problems of expensive, cumbersome, and low detection of the instrument, and achieve the effects of avoiding the influence of subjective factors, good repeatability, and fast and simple operation.

Inactive Publication Date: 2016-01-13
UNIV OF JINAN
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, RIA requires radioactive operation, and the price of the instrument is expensive, which has inspired scientists to explore non-radioactive-labeled immunoassay methods, and rapidly developed enzyme-linked immunoassay (ELISA), luminescence immunoassay (LIA), electrochemical Immunoassay (ECIA) and other methods
However, these immunoassay methods generally take a long time and have cumbersome operations such as adding samples, incubating, and washing, and the operators have more contact with immune substances, which may cause harm to the health of the operators.
Moreover, some detection methods with fast determination speed, high sensitivity and low detection performance require expensive instruments and cannot be popularized in developing countries.

Method used

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  • Preparation and applications of electrochemical immunosensor used for detecting circulating tumor cells

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preparation example Construction

[0032] (1) Preparation of vanadium pentoxide nanowires: It is characterized by mixing water-soluble vanadyl sulfate and oxidant potassium bromate at room temperature, and reacting with magnetic stirring for 30 minutes, and then reducing the acidity with nitric acid, and the aqueous solution of the mixture is heated in an autoclave at 180 React at ℃ for 24 hours, take out the autoclave and cool to room temperature, rinse with secondary water several times to get a dark yellow precipitate, dry the precipitate in an oven at 80°C for later use;

[0033] (2) Preparation of gold nanoparticle stock solution: it is characterized by adding 1.25mL of 10mM chloroauric acid into 42.65mL of secondary water, placing it in a three-neck round-bottomed flask with a capacity of 100mL, heating it for 30min under magnetic stirring, and then adding 2.5 mL of 1% sodium citrate, within 10 minutes, the solution first changed from yellow to blue-gray, and finally to soft pink. The solution was centrifu...

Embodiment 1

[0039] Example 1 (breast cancer cells, MCF-7)

[0040] (1) Select MCF-7 that is easy to transfer with the lymphatic system for determination;

[0041] (2) Preparation of vanadium pentoxide nanowires: Mix water-soluble vanadyl sulfate and oxidant potassium bromate at room temperature, and react with magnetic stirring for 30 minutes, then reduce the acidity with nitric acid, and react the aqueous solution of the mixture in an autoclave at 180°C for 24 hours , take out the autoclave and cool to room temperature, rinse with secondary water several times to get a dark yellow precipitate, dry the precipitate in an oven at 80°C for later use;

[0042] (3) Preparation of gold nanoparticle stock solution: add 1.25mL of 10mM chloroauric acid to 42.65mL of secondary water, place in a 100mL three-necked round-bottomed flask, heat for 30min under magnetic stirring, and then add 2.5mL of 1% Sodium citrate, the solution first changed from yellow to blue-gray within 10 minutes, and finally...

Embodiment 2

[0049] Example 2 (breast cancer cells, T47D)

[0050] (1) Select T47D that is easy to transfer with the lymphatic system for determination;

[0051] (2) Preparation of vanadium pentoxide nanowires: Mix water-soluble vanadyl sulfate and oxidant potassium bromate at room temperature, and react with magnetic stirring for 30 minutes, then reduce the acidity with nitric acid, and react the aqueous solution of the mixture in an autoclave at 180°C for 24 hours , take out the autoclave and cool to room temperature, rinse with secondary water several times to get a dark yellow precipitate, dry the precipitate in an oven at 80°C for later use;

[0052] (3) Preparation of gold nanoparticle stock solution: add 1.25mL of 10mM chloroauric acid to 42.65mL of secondary water, place in a 100mL three-necked round-bottomed flask, heat for 30min under magnetic stirring, and then add 2.5mL of 1% Sodium citrate, the solution first changed from yellow to blue-gray within 10 minutes, and finally t...

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Abstract

The invention discloses a sensor used for detecting circulating tumor cells via differential pulse voltammetry. A preparation method of the sensor comprises following steps: vanadium pentoxide nanowires and gold-silver core-shell nanoparticles are prepared via a conventional method; the vanadium pentoxide nanowires and gold-silver core-shell nanoparticles are combined so as to modify aptamers; the surface of an electrode is modified with gold nanoparticles, and then the modified electrode is connected with the vanadium pentoxide nanowire gold-silver core-shell nanoparticle modified thiolation aptamers; the obtained electrochemical sensor is used for detecting circulating tumor cells via combination with an electrochemical workstation; and the electrochemical workstation is used for detecting. The sensor is high in specificity and sensitivity; operation is simple; and detection limit is low.

Description

technical field [0001] The invention relates to the technical field of cell detection, and more specifically relates to a reduction current generated by semiconductor catalysis, and an alloy enhances the reduction current to detect the number of circulating tumor cells, which belongs to the preparation technology of an ultrasensitive electrochemical cell immune sensor. Background technique [0002] In recent years, immunoassay is an important means of diagnosis and treatment of tumor diseases in clinical examination. Malignant tumors, that is, cancer, have always been a serious threat to human health and life safety. Cancer cells refer to a new organism produced by malignant transformation and clonal proliferation of some cells with division potential under the action of carcinogenic factors. In addition to their own uncontrolled growth, cancer cells can also invade the surrounding normal tissues and even transfer to other parts of the body through the body's circulatory sy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/48G01N33/574B82Y30/00B82Y40/00
Inventor 颜梅邓文平于京华葛慎光刘海云张彦申蕾王秀杨春蕾朱少军杨光鑫
Owner UNIV OF JINAN
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