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Electrochemical immunodetection method

An immunodetection method and electrochemical technology are applied in the field of electrochemical immunodetection of thionine-graphene nanocomposite, which can solve the problem that it is difficult to detect trace abnormal prothrombin, affects the accuracy of primary liver cancer, and has low sensitivity. question

Inactive Publication Date: 2011-10-26
THE THIRD AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIV OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the low sensitivity of ELISA, it is difficult to detect trace amounts of abnormal prothrombin (APT), which affects the accuracy of primary liver cancer detection.

Method used

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  • Electrochemical immunodetection method

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Experimental program
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Effect test

Embodiment 1

[0046] The preparation of the nanocomposite of embodiment 1 nano-platinum chitosan double wrapping thionine-graphene

[0047] Add 5 mg of thionine to 4.5 mL of graphene aqueous solution with a mass fraction of 0.05%, continue ultrasonication at 20° C. for 4 h, and centrifuge and wash with ethanol and deionized water, respectively, to remove thionine that is not combined with graphene, and obtain thionine- Graphene composite: Add the thionine-graphene composite to 5.5mL of 10mg / mL chitosan solution, stir continuously at 60°C for 12h, add 30μL of 0.1mol / L acetic acid dropwise, and stir at room temperature for 10min , continue heating in an oil bath at 90° C. for 1 h, and filter with a 0.2 μm nylon membrane to obtain chitosan-wrapped thionine-graphene nanoparticles; the chitosan-wrapped thionine-graphene nanoparticles Ultrasonic dispersion in water, take 5 mg and 80 μL of 1% HPtCl 6 Mix well and add to 10mL water, stir vigorously for 5min, add 0.25mL 100mmol / L NaBH dropwise 4 ,...

Embodiment 2

[0048] The preparation of the nanocomposite of embodiment 2 nano-platinum chitosan double coating thionine-graphene

[0049]Add 5 mg of thionine to 4.5 mL of graphene aqueous solution with a mass fraction of 0.05%, continue ultrasonication at 20° C. for 4 h, and centrifuge and wash with ethanol and deionized water, respectively, to remove thionine that is not combined with graphene, and obtain thionine- Graphene composite: Add the thionine-graphene composite to 5.5mL of 10mg / mL chitosan solution, stir continuously at 60°C for 12h, add 30μL of 0.1mol / L acetic acid dropwise, and stir at room temperature for 10min , continue heating in an oil bath at 90° C. for 1 h, and filter with a 0.26 μm nylon membrane to obtain thionine-graphene nanoparticles wrapped in chitosan; the thionine-graphene nanoparticles wrapped in chitosan Ultrasonic dispersion in water, take 5 mg and 120 μL of 1% HPtCl 6 Mix well and add to 10mL water, stir vigorously for 5min, add 0.25mL 100mmol / L NaBH dropwis...

Embodiment 3

[0050] The preparation of the nanocomposite of embodiment 3 nano-platinum chitosan double coating thionine-graphene

[0051] Add 5 mg of thionine to 4.5 mL of graphene aqueous solution with a mass fraction of 0.05%, continue ultrasonication at 20° C. for 4 h, and centrifuge and wash with ethanol and deionized water, respectively, to remove thionine that is not combined with graphene, and obtain thionine- Graphene composite: Add the thionine-graphene composite to 5.5mL of 10mg / mL chitosan solution, stir continuously at 60°C for 12h, add 30μL of 0.1mol / L acetic acid dropwise, and stir at room temperature for 10min , continue heating in an oil bath at 90° C. for 1 h, and filter with a 0.22 μm nylon membrane to obtain thionine-graphene nanoparticles wrapped in chitosan; the thionine-graphene nanoparticles wrapped in chitosan Ultrasonic dispersion in water, take 5 mg and 150 μL of 1% HPtCl 6 Mix well and add to 10mL water, stir vigorously for 5min, add 0.25mL 100mmol / L NaBH dropwi...

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Abstract

The invention discloses an electrochemical immunodetection method based on a thionine-graphene nano composite double coated with nano platinum chitosan. According to the invention, double antibody sandwiched method is adopted, an abnormal prothrombin(APT) antibody supported on the surface of electrodes is subject to an immunization reaction with an abnormal prothrombin(APT) in a sample solution, then combines with the coupling object of the thionine-graphene nano composite double coated with nano platinum chitosan and the abnormal prothrombin(APT). Based on the electrochemical activity of thethionine-graphene nano composite double coated with nano platinum chitosan, peak current value of reduction peak of cyclic voltammetry is detected so as to further detect the concentration of the abnormal prothrombin in the sample under test. The method has the linear response range of 0.8-800ng / ml, the lower limit of the detection of 0.23ng / ml, with good specificity and high sensitivity, and is significant to the diagnosis of the abnormal prothrombin(APT).

Description

technical field [0001] The invention relates to the field of electrochemical immunity, in particular to an electrochemical immunodetection method based on a thionine-graphene nanocomposite double-wrapped by nano-platinum chitosan. Background technique [0002] Primary liver cancer (primary carcinoma of the liver) is one of the common malignant tumors in my country. The mortality rate is high, ranking third after gastric cancer and esophageal cancer in the mortality rate of malignant tumors, and ranking second in rural areas in some areas, second only to gastric cancer. About 110,000 people die from liver cancer every year in my country, accounting for 45% of the world's liver cancer deaths. Therefore, it is particularly important to make an early diagnosis of primary liver cancer, formulate a reasonable treatment plan, and evaluate the curative effect. At present, the main methods used for the auxiliary diagnosis of primary liver cancer are chest X-ray, CT, and sputum cyto...

Claims

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Application Information

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IPC IPC(8): G01N27/26G01N27/327G01N21/76G01N33/68B82Y40/00
Inventor 仲召阳卿毅王东李梦侠戴楠关伟单锦露
Owner THE THIRD AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIV OF PLA
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