Electro-Nano Diagnostic Platform Based on Antibody–Antigen Interaction: An Electrochemical Immunosensor for Influenza A Virus Detection

H1N1 is a kind of influenza A virus that causes serious health issues throughout the world. Its symptoms are more serious than seasonal flu and can sometimes be lethal. For this reason, rapid, accurate, and effective diagnostic tests are needed. In this study, an electrochemical immunosensor for the sensitive, selective, and practical detection of the H1N1 virus was developed. The sensor platform included multi-walled carbon nanotube gold-platinum (MWCNT-Au-Pt) hybrid nanomaterial and anti-hemagglutinin (anti-H1) monoclonal antibody. For the construction of this biosensor, a gold screen-printed electrode (AuSPE) was used as a transducer. Firstly, AuSPE was modified with MWCNT-Au-Pt hybrid nanomaterial via drop casting. Anti-H1 antibody was immobilized onto the electrode surface after the modification process with cysteamine was applied. Then, the effect of the interaction time with cysteamine for surface modification was investigated. Following that, the experimental parameters, such as the amount of hybrid nanomaterial and the concentration of anti-H1 were optimized. Under the optimized conditions, the analytical characteristics of the developed electrochemical immunosensor were investigated for the H1N1 virus by using electrochemical impedance spectroscopy. As a result, a linear range was obtained between 2.5–25.0 µg/mL with a limit of the detection value of 3.54 µg/mL. The relative standard deviation value for 20 µg/mL of the H1N1 virus was also calculated and found as 0.45% (n = 3). In order to determine the selectivity of the developed anti-H1-based electrochemical influenza A immunosensor, the response of this system towards the H3N2 virus was investigated. The matrix effect was also investigated by using synthetic saliva supplemented with H1N1 virus.

[1]  K. Kuronuma,et al.  Comparative study of rapid antigen testing and two nucleic acid amplification tests for influenza virus detection. , 2022, Journal of infection and chemotherapy : official journal of the Japan Society of Chemotherapy.

[2]  B. Cowling,et al.  Human seasonal influenza under COVID-19 and the potential consequences of influenza lineage elimination , 2021, Nature Communications.

[3]  Yudum Tepeli Büyüksünetçi,et al.  An impedimetric approach for COVID-19 detection. , 2021, The Analyst.

[4]  Ülkü Anık,et al.  Development and application of a SARS-CoV-2 colorimetric biosensor based on the peroxidase-mimic activity of γ-Fe2O3 nanoparticles , 2021, Microchimica Acta.

[5]  L. Brammer,et al.  Changes in Influenza and Other Respiratory Virus Activity During the COVID-19 Pandemic — United States, 2020–2021 , 2021, MMWR. Morbidity and mortality weekly report.

[6]  L. Brammer,et al.  Decreased influenza activity during the COVID-19 pandemic—United States, Australia, Chile, and South Africa, 2020 , 2020, American Journal of Transplantation.

[7]  Moustafa Almunla,et al.  Development of Apple Tissue Based Biocathode and MWCNT−Pt−Au Nanomaterial Based Bioanode Biofuel Cell , 2020 .

[8]  David W. Smith,et al.  Where has all the influenza gone? The impact of COVID-19 on the circulation of influenza and other respiratory viruses, Australia, March to September 2020 , 2020, Euro surveillance : bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin.

[9]  Ravina,et al.  Genosensor for rapid, sensitive, specific point-of-care detection of H1N1 influenza (swine flu) , 2020 .

[10]  L. Brammer,et al.  Decreased influenza activity during the COVID-19 pandemic—United States, Australia, Chile, and South Africa, 2020 , 2020, American Journal of Transplantation.

[11]  P. Karimian,et al.  Comparative study of clinical symptoms, laboratory results and imaging features of coronavirus and influenza virus, including similarities and differences of their pathogenesis , 2020 .

[12]  S. Yao,et al.  A sensitive electrochemical sensor for bisphenol A on the basis of the AuPd incorporated carboxylic multi-walled carbon nanotubes. , 2019, Food chemistry.

[13]  Akira Matsumoto,et al.  Human influenza virus detection using sialyllactose-functionalized organic electrochemical transistors , 2018 .

[14]  Ying-Feng Chang,et al.  Simple Strategy for Rapid and Sensitive Detection of Avian Influenza A H7N9 Virus Based on Intensity-Modulated SPR Biosensor and New Generated Antibody. , 2018, Analytical chemistry.

[15]  D. Pang,et al.  Digital Single Virus Electrochemical Enzyme-Linked Immunoassay for Ultrasensitive H7N9 Avian Influenza Virus Counting. , 2018, Analytical chemistry.

[16]  S. Kaushik,et al.  Evaluation of clinical applicability of reverse transcription-loop-mediated isothermal amplification assay for detection and subtyping of Influenza A viruses , 2017, Journal of Virological Methods.

[17]  Yudum Tepeli,et al.  Electrochemical biosensors for influenza virus a detection: The potential of adaptation of these devices to POC systems , 2018 .

[18]  W. Goddard,et al.  A rapid-response ultrasensitive biosensor for influenza virus detection using antibody modified boron-doped diamond , 2017, Scientific Reports.

[19]  B. Muszyńska,et al.  Artificial saliva and its use in biological experiments. , 2017, Journal of physiology and pharmacology : an official journal of the Polish Physiological Society.

[20]  Ronghui Wang,et al.  A nanowell-based QCM aptasensor for rapid and sensitive detection of avian influenza virus , 2017 .

[21]  M. Diouani,et al.  Towards the electrochemical diagnostic of influenza virus: development of a graphene-Au hybrid nanocomposite modified influenza virus biosensor based on neuraminidase activity. , 2017, The Analyst.

[22]  M. Diouani,et al.  Fabrication of Electrochemical Model Influenza A Virus Biosensor Based on the Measurements of Neuroaminidase Enzyme Activity. , 2016, Analytical chemistry.

[23]  O. Liesenfeld,et al.  Performance of the Cobas® Influenza A/B Assay for Rapid Pcr-Based Detection of Influenza Compared to Prodesse ProFlu+ and Viral Culture , 2015, European journal of microbiology & immunology.

[24]  D. Dwyer,et al.  Detection of influenza A and B with the Alere™ i Influenza A & B: a novel isothermal nucleic acid amplification assay , 2015, Influenza and other respiratory viruses.

[25]  Ey,et al.  Surface Orchestration of Gold Nanoparticles Using Cysteamine as Linker and Folate as Navigating Molecule for Synaphic Delivery of Doxorubicin , 2014 .

[26]  Kai Ludwig,et al.  Receptor binding and pH stability - how influenza A virus hemagglutinin affects host-specific virus infection. , 2014, Biochimica et biophysica acta.

[27]  G. Gao,et al.  Bat-derived influenza-like viruses H17N10 and H18N11 , 2014, Trends in Microbiology.

[28]  Yingchun Fu,et al.  Exploiting enzyme catalysis in ultra-low ion strength media for impedance biosensing of avian influenza virus using a bare interdigitated electrode. , 2014, Analytical chemistry.

[29]  J. Pedersen Hemagglutination-inhibition assay for influenza virus subtype identification and the detection and quantitation of serum antibodies to influenza virus. , 2014, Methods in molecular biology.

[30]  Hua Yang,et al.  New World Bats Harbor Diverse Influenza A Viruses , 2013, PLoS pathogens.

[31]  J. Katz,et al.  Immunological assessment of influenza vaccines and immune correlates of protection , 2013, Expert review of vaccines.

[32]  S. E. Diltemiz,et al.  4-Aminophenyl boronic acid modified gold platforms for influenza diagnosis. , 2013, Materials science & engineering. C, Materials for biological applications.

[33]  A. K. Al-zobaei Comparison between Haemagglutination Inhibition and Complement Fixation Tests in Detecting Antibodies Responses Following Influenza Viral Infection , 2012 .

[34]  G. Fischer,et al.  Performance of five FDA‐approved rapid antigen tests in the detection of 2009 H1N1 influenza A virus , 2012, Journal of medical virology.

[35]  N. Sriwilaijaroen,et al.  Molecular basis of the structure and function of H1 hemagglutinin of influenza virus , 2012, Proceedings of the Japan Academy. Series B, Physical and biological sciences.

[36]  S. Shahrokhian,et al.  Construction of an electrochemical sensor based on the electrodeposition of Au-Pt nanoparticles mixtures on multi-walled carbon nanotubes film for voltammetric determination of cefotaxime. , 2012, The Analyst.

[37]  T. Mahmood,et al.  Influenza A H1N1 2009 (Swine Flu) and Pregnancy , 2011, Journal of obstetrics and gynaecology of India.

[38]  Wei Wang,et al.  Nanoparticle-based biosensor for the detection of emerging pandemic influenza strains. , 2010, Biosensors & bioelectronics.

[39]  U. Liebert,et al.  Swine-origin H1N1 influenza A virus and dental practice: a critical review , 2010, Clinical Oral Investigations.

[40]  M. Oyama,et al.  Effects of linker molecules on the attachment and growth of gold nanoparticles on indium tin oxide surfaces , 2009 .

[41]  G. Tannock,et al.  The detection of influenza A and B viruses in clinical specimens using a quartz crystal microbalance , 2009, Journal of Virological Methods.

[42]  Ron A M Fouchier,et al.  Antigenic and Genetic Characteristics of Swine-Origin 2009 A(H1N1) Influenza Viruses Circulating in Humans , 2009, Science.

[43]  M. Ciotti,et al.  Origin of the 2009 Mexico influenza virus: a comparative phylogenetic analysis of the principal external antigens and matrix protein , 2009, Archives of Virology.

[44]  Xin Wang,et al.  Graphene−Metal Particle Nanocomposites , 2008 .

[45]  N. Jaffrezic‐Renault,et al.  Miniaturized biosensor for avian influenza virus detection , 2008 .

[46]  Marta Ligaj,et al.  Application of DNA Hybridization Biosensor as a Screening Method for the Detection of Genetically Modified Food Components , 2008, Sensors.

[47]  C. Naeve,et al.  Large-Scale Sequence Analysis of Avian Influenza Isolates , 2006, Science.

[48]  James C Paulson,et al.  Glycan microarray analysis of the hemagglutinins from modern and pandemic influenza viruses reveals different receptor specificities. , 2006, Journal of molecular biology.

[49]  Jing-Juan Xu,et al.  A glucose biosensor based on chitosan-glucose oxidase-gold nanoparticles biocomposite formed by one-step electrodeposition. , 2004, Analytical biochemistry.

[50]  H. Aizawa,et al.  Differences in clinical features between influenza A H1N1, A H3N2, and B in adult patients , 2003, Respirology.

[51]  J. Taubenberger,et al.  The 1918 Spanish influenza: integrating history and biology. , 2001, Microbes and infection.

[52]  L. Mitnaul,et al.  Balanced Hemagglutinin and Neuraminidase Activities Are Critical for Efficient Replication of Influenza A Virus , 2000, Journal of Virology.

[53]  Leif Nyholm,et al.  Self-Assembled Monolayers of Cystamine and Cysteamine on Gold Studied by XPS and Voltammetry , 1999 .

[54]  A. Steel,et al.  Electrochemical quantitation of DNA immobilized on gold. , 1998, Analytical chemistry.

[55]  T. M. Herne,et al.  Characterization of DNA Probes Immobilized on Gold Surfaces , 1997 .

[56]  R. Lamb,et al.  Orthomyxoviridae: The Viruses and Their Replication. , 1996 .

[57]  W. S. Hummers,et al.  Preparation of Graphitic Oxide , 1958 .