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Biosensor, preparation method and application thereof, and electrochemical system for detecting ctDNA

A biosensor and system technology, applied in the field of ctDNA detection, can solve the problems of large instruments and unfavorable portability of fluorescence detection methods

Active Publication Date: 2022-02-25
YUNNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Fluorescence detection requires a larger instrument, which is not conducive to portability

Method used

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  • Biosensor, preparation method and application thereof, and electrochemical system for detecting ctDNA
  • Biosensor, preparation method and application thereof, and electrochemical system for detecting ctDNA
  • Biosensor, preparation method and application thereof, and electrochemical system for detecting ctDNA

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preparation example Construction

[0075] In a specific embodiment of the present invention, when the magnetic covalent organic framework is Fe 3 o 4 @COF, the Fe 3 o 4 The preparation method of @COF preferably includes the following steps:

[0076] Ferric chloride, ethylene glycol, sodium acetate and polyethylene glycol were mixed for solvothermal reaction to obtain Fe 3 o 4 nanospheres;

[0077] the Fe 3 o 4 After the nanospheres, benzidine and tetrahydrofuran (THF) are mixed and refluxed, a tetrahydrofuran solution of trialdehyde phloroglucinol (TP) is added to carry out a template-mediated precipitation reaction to obtain the Fe 3 o 4 @COF.

[0078] The present invention mixes ferric chloride, ethylene glycol, sodium acetate and polyethylene glycol, and carries out solvothermal reaction to obtain Fe 3 o 4 nanospheres.

[0079] In the present invention, the ferric chloride is preferably FeCl 3 .6H 2 O.

[0080] In the present invention, the dosage ratio of the ferric chloride, ethylene glycol,...

Embodiment 1

[0150] Take 1.35g FeCl 3 .6H 2 O mixed with 40mL ethylene glycol, added 3.6g sodium acetate and 1.0g polyethylene glycol, stirred for 0.5h, solvothermal reaction at 180°C for 6h, cooled to room temperature, washed several times with ethanol, dried at 60°C for 4h to obtain Fe 3 o 4 (nanospheres);

[0151] Take 32mg of said Fe 3 o 4 and 22 mL of THF solution containing 32 mg of BD, sonicated for 15 min, stirred and refluxed at 50°C for 0.5 h. Then add 8 mL of THF solution containing 24 mg TP dropwise at a rate of 8 drops / min, stir at 50°C for 12 hours, and filter with suction to obtain Fe 3 o 4 @COF;

[0152] 10mg of the Fe 3 o 4 @COF, 10mgAu-Pd and 10mL deionized water were mixed, after ultrasonic dispersion, 0.1mg methylene blue was added, stirred at room temperature for 24h, and the precipitate was collected after centrifugation to obtain Fe 3 o 4 @COF / Pd-Au / methylene blue;

[0153] 10mg Na 2 PdCl 4 , 2.1mg HAuCl 4 , 30mg W(CO) 6 After mixing with 2 mL of acet...

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Abstract

The invention relates to the technical field of ctDNA detection, in particular to a biosensor, a preparation method and application thereof and an electrochemical system for detecting ctDNA. The sensor comprises a substrate electrode, an Au-CP1 probe layer, an HT layer, a CRSPR / Cas12a system layer and a CP2 probe / magnetic covalent organic framework / Pd-Au / MB layer, wherein the Au-CP1 probe layer, the HT layer, the CRSPR / Cas12a system layer and the CP2 probe / magnetic covalent organic framework / Pd-Au / MB layer are sequentially loaded on the surface of the substrate electrode; the CRSPR / Cas12a system layer comprises crRNA, Cas12a, ssDNA and a sample to be detected; the CP2 probe / magnetic covalent organic framework / Pd-Au / MB layer comprises a magnetic covalent organic framework and Pd-Au and MB which are loaded in a pore structure of the magnetic covalent organic framework, and further comprises a CP2 probe; and Au in the Pd-Au is bonded with sulfydryl in the CP2 probe. The biosensor is convenient to carry and can specifically detect ctDNA.

Description

technical field [0001] The invention relates to the technical field of ctDNA detection, in particular to a biosensor, a preparation method and application thereof, and an electrochemical system for detecting ctDNA. Background technique [0002] Lung cancer is currently recognized as a malignant tumor with high morbidity and mortality in the world, so accurate and rapid early diagnosis is of great significance for the clinical treatment and prognosis of lung cancer patients. About 80% of lung cancer is non-small cell lung cancer (NSCLC), which has a high degree of malignancy and poor prognosis. Clinical studies have shown that more than 60% of NSCLC highly express epidermal growth factor receptor (EGFR), and EGFR has been considered as an important therapeutic target for the treatment of NSCLC. Most EGFR mutations occur in the tyrosine kinase domain in exons 18-21. Mutations in exon 21 (EGFR L858R) account for 40% to 45% of these mutations in most cases. EGFR L858R is high...

Claims

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Application Information

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IPC IPC(8): G01N27/26G01N27/30G01N27/327
CPCG01N27/26G01N27/30G01N27/3278
Inventor 赵卉李灿鹏张亚平刘凤
Owner YUNNAN UNIV
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