Research On Hantan Virus Coulometric Immunosensor Based On Printed Carbon Electrodes

People have been threatened by diseases caused by various viruses for so long.Hantaan virus(HTNV)is one of the viruses that threaten human health seriously.Hantaan virus infection mainly causes hemorrhagic fever with renal syndrome(HFRS)in China.The main clinical symptoms of HFRS are fever,hemorrhage,hyperemia,hypotension shock and kidney damage.This disease has a wide range of epidemics,a large number of cases,and a high rate of mortality.Up to the present day,there is no effective therapeutic drug in clinical practice,so HTNV infection early diagnosis is great significant for the treatment of HFRS.Virus-like particles(VLPs)are hollow particles containing the original viral structural proteins,which have good biosafety,no virulence,and no infection problems.The ectrochemical coulometric immunosensors use an antigen or an antibody as a molecular recognition component,and converts various signals change caused by the immune reaction into corresponding electrical signals through a signal converter,which own superiority of high precision,simple equipment,easy automation fast,high sensitivity,high selectivity and low cost.Electrochemical coulometric immunosensor have attracted more and more attention in virus detection In this paper,Screen-printed carbon electode(SPCE)is used as the working electrode,antibodies are used as molecular recognition substances,HTNV VLPs and IgM are the targets,and Horseradish Peroxidase(HRP)is used as the marker,to bulid a high-sensitivity Coulomb-type immunosensor for detecting HTNV and provided a criterion for the diagnosis of serum HTNV infection.This paper consists of three parts.Chapter 1 is the introduction.This chapter first gives a brief overview of HTNV,including its clinical manifestations and present detection methods,also introduce the virus-like particles,paticularly the application of virus-like particles were illustrated.Followed by a detailed introduction of biosensors and electrochemical immunosensors,especially for biosensors and electrochemical immunosensors.Then introduce the screen printing technology and its application in virus detection;followed by a brief overview of the electron mediator;Chapter 2 is a study for applying Coulomb-type electrochemical immunosensor to detect Hantaan virus,based on screen-printed carbon electrodes.First of all,the surface of the SPCE electrode is functionalized by electropolymerization,and then the 1A8 antibody is covalently bonded on the surface of the electrode by activating the electrode with glutaraldehyde.HTNV VLPs and HRP-labeled 1A8 antibodies were subsequently immobilized on the electrode surface by specific recognition.After that,by electrochemically depositing a layer of ruthenium-containing macromolecules on the surface of the electrode as an electron mediator,the charge is accumulated at a constant potential of-0.1 V.The purpose of detecting HTNV is based on the relationship between the amount of reduction in 1200 s and the concentration of HTNV VLPs on the surface of the electrode.The experimental results show that the response of the SPCE Coulometric immunosensor has a good linear relationship with the concentration of HTNV VLPs in the range of 100 pfu mL-1 to 10000 pfu·mL-1.The detection limit is 10 pfu·L-1.Chapter 3 is the early diagnosis of Hantaan virus infection by Coulomb-type electrochemical immunosensors for.This experiment is based on the ELISA commonly used in clinical diagnosis,and aims to provide a criterion for the determination of the Coulomb type immunosensor.In this study,the surface of the SPCE electrode was functionalized by electropolymerization,and activated by glutaraldehyde to immobilized M12 antibody,which specifically recognizes IgM in serum when the serum is positive,there is an IgM in the serum that can connected with HTNV The IgM captures the HTNV VLPs in the solution,and further fixes the HRP-labeled 1A8 antibody to the surface of the electrode by immunospecific recognition,which will reduces the H2O2 in the catalytic solution,and outputs a charge signal.The negative serum sample absent this IgM,so there is no 1A8-HRP on the electrode surface to change the signal.Therefore,the reduce power of the positive serum sample in 1200s is greater than the negative serum sample,and there is a relatively fixed value between them.The experimental results show that the value of 1.72 can use as a criterion,to judge whether the serum supplier is infected with HTNV or not.As for the practical sample,when the ratio of unknown serum to negative serum-like electricity is bigger than this value,the serum sample is positive,vice versa.This immunosensor provides a new method for early diagnosis of HTNV infection.