CN-120253998-B - COF-based HHV-8 detection electrochemical sensor, preparation method and application

Abstract

The invention belongs to the field of virus detection, and particularly relates to a COF-based HHV-8 detection electrochemical sensor, a preparation method and application. The preparation method comprises the steps of dissolving TMACL and KTPBCl in a solvent according to an equimolar ratio of 1:1, stirring for reaction, centrifuging to obtain precipitate, drying to obtain TMATPBCl, mixing PDA, TAPB and TMATPBCl, adding into DCE and the solvent, standing to obtain mixed solution, taking out a COF film, washing, performing ultrasonic dispersion to obtain a COF dispersion material, shearing carbon paper into a size of 1cm multiplied by 1cm for standby, sequentially performing ultrasonic treatment on a clamping piece electrode in water, absolute ethyl alcohol and water, clamping the carbon paper after the clamping piece electrode is dried, coating the COF dispersion material obtained in S2 on the surface of the carbon paper electrode, performing preservation and drying, and dripping HHV8 antibody to obtain the COF detection HHV8 electrochemical immunosensor.

Inventors

• CHEN MINGXIN
• SHANG SHUYONG
• Dai Tenglong
• LI JIAGEN
• HE ZIHANG
• QIU SI
• DU YAFEI

Assignees

• 成都师范学院

Dates

Publication Date

20260505

Application Date

20250408

Claims (3)

1. 1. The preparation method of the HHV-8 electrochemical sensor based on COF detection is characterized by comprising the following steps: s1, dissolving TMACL and KTPBCl in a solvent in an equimolar ratio of 1:1, stirring for reaction, centrifuging to obtain precipitate, and drying to obtain TMATPBCl, wherein the solvent in S1 is a mixed solution of methanol and water, and the volume ratio of the methanol to the water is 2:1; S2, mixing TMATPBCl obtained by PDA, TAPB and S1, adding into DCE and a solvent, standing for 24: 24h at the temperature of 2-8 ℃ to obtain a COF film, taking out the COF film, flushing the COF film by using absolute ethyl alcohol, adding the absolute ethyl alcohol after flushing, and performing ultrasonic dispersion to obtain a COF dispersion material, wherein the solvent in S2 is formed by mixing water, TMACL and concentrated hydrochloric acid; The molar ratio of PDA, TAPB and TMATPBCl is 1:1.5:1; And S3, cutting the carbon paper into the size of 1 cm multiplied by 1 cm for standby, sequentially carrying out ultrasonic treatment on the clamping sheet electrode in water, absolute ethyl alcohol and water, clamping the carbon paper after the clamping sheet electrode is dried, coating the COF dispersion material obtained in the S2 on the surface of the carbon paper electrode, preserving and drying, and dripping the HHV-8 antibody to obtain the COF detection HHV-8 electrochemical immunosensor.
2. 2. The method for preparing the COF-based HHV-8 electrochemical sensor according to claim 1, wherein the preservation time in S3 is 4 h and the preservation temperature is 4-8 ℃.
3. 3. A COF-based HHV-8 electrochemical sensor obtained by the preparation method according to any one of claims 1 to 2.

Description

COF-based HHV-8 detection electrochemical sensor, preparation method and application Technical Field The invention belongs to the field of virus detection, and particularly relates to a COF-based HHV-8 detection electrochemical sensor, a preparation method and application. Background Human herpesvirus type 8 (HHV-8) is one of the subfamilies of herpesvirus gamma. The virus is the causative agent of Kaposi's Sarcoma (KS), a cancerogenic disease that has a great impact on immunocompromised individuals, and in addition, HHV-8 has been closely related to a wide range of clinical diseases, with recent research efforts revealing its potential association with multicenter Kaposi's disease (MCD), a life threatening, virally induced b-lymphoproliferative disease. This clearly further widens its relevant scope in clinical medicine. Thus, the importance of HHV-8 infection is both critical and urgent. Currently, real-time fluorescent quantitative PCR technology has become the dominant means for detecting HHV-8 infection. The technique allows accurate quantification of viral DNA and transcript levels in blood, as well as other body fluids and organs. However, it should be ignored that part of the detection process still appears to be complex, cumbersome, time-consuming and of limited sensitivity. During the HHV-8 latent infection phase, antigens of the relevant viruses are stably expressed, which property makes them a marker biomarker for monitoring latent HHV-8 infection. This finding provides a powerful support for early diagnosis and scientific monitoring of HHV-8 related diseases. In view of this, it is particularly urgent and important to develop a novel, highly sensitive, rapid and cost-effective detection method, specifically for the detection of antigens in HHV-8. Disclosure of Invention Aiming at the problems and disadvantages of the prior art, the invention provides a preparation method of an HHV-8 electrochemical sensor based on COF detection, which comprises the following steps: S1, dissolving TMACL and KTPBCl in a solvent in an equimolar ratio of 1:1, stirring for reaction, centrifuging to obtain a precipitate, and drying to obtain TMATPBCl; S2, mixing TMATPBCl obtained by PDA, TAPB and S1, adding into DCE and solvent, standing to obtain mixed solution, taking out the COF film, washing, and performing ultrasonic dispersion to obtain a COF dispersion material; The molar ratio of PDA, TAPB and TMATPBCl is 1:1.5:1; And S3, cutting the carbon paper into the size of 1cm multiplied by 1cm for standby, sequentially carrying out ultrasonic treatment on the clamping piece electrode in water, absolute ethyl alcohol and water, clamping the carbon paper after the clamping piece electrode is dried, coating the COF dispersion material obtained in the S2 on the surface of the carbon paper electrode, preserving and drying, and dripping the HHV8 antibody to obtain the COF detection HHV8 electrochemical immunosensor. Further, the solvent in S1 is a mixed solution of methanol and water, and the volume ratio of the methanol to the water is 2:1. Further, the standing time in the step S2 is 24 hours, the standing temperature is 2-8 ℃, and absolute ethyl alcohol is used in the ultrasonic dispersion process. Further, the preservation time in S3 is 4 hours, and the preservation temperature is 4-8 ℃. An HHV-8 electrochemical sensor based on COF detection. Use of a COF-based detection HHV-8 electrochemical sensor for the preparation of a device for detecting HHV-8 type human herpesvirus. Advantageous effects The invention provides a preparation method of a HHV-8 electrochemical sensor based on COF detection, which successfully utilizes a hydrophilic TAPB-PDA COF film with high mechanical strength and carbon paper with excellent conductivity, the porous structure of the COF provides high specific surface area loaded antibodies, and the rapid electronic transmission of amplified signals of a carbon paper electrode, so that the ultra-sensitive detection of HHV-8 virus antigen is realized by the cooperation of the hydrophilic TAPB-PDA COF film and the carbon paper. This achievement benefits from the fact that the amino groups in the material have a hydrophilic character that effectively binds to the carboxyl groups of the HHV-8 antibody. The designed biological immune biosensor shows excellent performance indexes, namely, the detection range is wide, the linear range from 4.97X10 -16 g/mL to 3.68X10 -13 g/mL is covered, the detection sensitivity is extremely high, the detection limit is as low as 0.142X 10 -16 g/mL (signal to noise ratio S/N=3), and is far lower than the fluorescent quantitative PCR method commonly used in the current detection and is also lower than the detection limit (more than or equal to 10 -13 g/mL) of the current most electrochemical sensors. And the molar ratio of DA, TAPB and TMATPBCl is 1:1.5:1, so that the electrochemical sensor has better effect compared with the prior art. In the prior art, fe 3O4 @COF is ad