Document Type : Original Paper
Authors
1 Mechanical Engineering of Biosystems Department, Faculty of Agriculture, Shahrekord University, 8818634141, Shahrekord, Islamic Republic of Iran
2 1 Mechanical Engineering of Biosystems Department, Faculty of Agriculture, Shahrekord University, 8818634141, Shahrekord, Islamic Republic of Iran 2 Nanotechnology Research Center, Shahrekord University, 8818634141, Shahrekord, Islamic Republic of Iran
3 Department of Genetics, Faculty of Basic Science, Shahrekord University, 8818634141, Shahrekord, Islamic Republic of Iran
4 Cellular & Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences (SKUMS), Shahrekord, Islamic Republic of Iran
Abstract
Nowadays the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has become an endemic disease throughout the world on the other hand intensive worldwide vaccination programs decreased the severity of the affection but early virus detection and disease diagnosis are still important healthcare management of infectious disease control. Therefore, in this research, we introduce an electrochemical genosensor based on a DNA probe that can hybridize directly to the viral genome or its transcripts and therefore does not need cDNA synthesis following RNA extraction from patient samples, a necessary and challenging step in routine RNA virus detection methods like Real-time PCR. Altogether, in this research, an electrochemical biosensor based on a virus-specific probe with thiol modification was designed and immobilization of the probe was carried out through self-assembly by thiol binding on reduced graphene oxide (RGO) and gold nanoparticles composite-modified pencil graphite electrode (PGE). The hybridizations of probe and target sequences were analyzed by electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) methods. The linear range was found to be within 10-12 - 10-6 M and the limit of detection (LOD) was at 3× 10 -13 M. The time of 20 minutes was chosen as the optimal hybridization time. The results showed that the fabricated biosensor can be recovered and reused up to 6 times. This means significant time, and expense savings when compared with other conventional detection methods for this virus. Therefore, this biosensor is suggested for clinical applications especially when time and sensitivity are the most limited elements.
Keywords
Main Subjects
- Eissa S. Diagnostic biosensors for coronaviruses and recent developments. Advanced Biosensors for Virus Detection. 2022; 8: 261-278. Doi: 10.1016/B978-0-12-824494-4.00008-4
- Moattari G, Izadi Z, Shakhsi-Niaei M. Development of an electrochemical genosensor for detection of viral hemorrhagic septicemia virus (VHSV) using glycoprotein (G) gene probe. Aquaculture. 2021; 536: 736451.
- Li J, Jin X, Feng M, Huang S, Feng J. Ulteransensitive and highly selective electrochemical biosensor for HIV gene detection based on Amino-reduced graphene oxide and b-cyclodextrin modified glassy carbon electrode. International Journal of Electrochemistry Science. 2020; 15: 2722-2738.
- Zhao F, Bai Y, Cao L, Han G, Fang C, Wei S, Chen Z. New electrochemical DNA sensor based on nanoflowers of Cu3 (PO4) 2-BSA-GO for hepatitis B virus DNA detection. Journal of Electroanalytical Chemistry. 2020; 867:114184.
- Chowdhury AD, Takemura K, Li T-C, Suzuki TM, Park EY. Electrical pulse-induced electrochemical biosensor for hepatitis E virus detection. Nat, Commun. 2019; 10: 3737.
- Lee T, Park SY, Jang H, Kim GH, Lee Y, Park C, Mohammadniaei M, Lee MH, Min J. Fabrication of electrochemical biosensor consisted of multi-functional DNA structure/porous au nanoparticle for avian influenza virus (H5N1) in chicken serum. Material Science Engineering. 2019; 99: 511-519.
- Faria HAM, Zucolotto V. Label-free electrochemical DNA biosensor for zika virus identification. Biosensors and Bioelectronics. 2019; 131: 149-155.
- Sabzi RE, Sehatnia B, Pournaghi-Azar MH, Hejazi MS. Electrochemical detection of human papilloma virus ( HPV ) target DNA using MB on pencil graphite electrode, . Journal of the Iranian Chemical Society. 2008; 5: 476–483. https://doi.org/10.1007/BF03246005
- Campos-Ferreira DS, Souza EVM, Nascimento GA, Zanforlin DML, Arruda MS, Beltrão MFS.et al. Electrochemical DNA biosensor for the detection of human papillomavirus E6 gene inserted in recombinant plasmid. Arabian Journal of Chemistry. 2016; 9: 443–450.
- Balvedi RPA, Castro ACH, Madurro JM, Brito-Madurro AG. Detection of a specific biomarker for epstein-barr virus using a polymer-based genosensor. International Journal of Molecular Sciences. 2014; 15: 9051–9066.
- Dong S, Zhao R, Zhu J, Lu X, Li Y Qiu S, et al. Electrochemical DNA Biosensor Based on a Tetrahedral Nanostructure Probe for the Detection of Avian Influenza A (H7N9) Virus. ACS Applied Material Interfaces. 2015; 7: 8834–8842.
- Shakoori Z, Salimian S, Kharrazi S, Adabi M, Saber R. Electrochemical DNA biosensor based on gold nanorods for detecting hepatitis B virus. Analytical and Bioanalytical Chemistry. 2015; 407: 455–461.
- Manzano M, Viezzi S, Mazerat S, Marks RS, Vidic J. Rapid and label-free electrochemical DNA biosensor for detecting Hepatitis A virus. Biosensors and Bioelectronics. 2018; 100: 89-95.
- Shawky SM, Awad AM, Allam W, Alkordi MH, EL-Khamisy SF. Gold aggregating gold: A novel nanoparticle biosensor approach for the direct quantification of hepatitis C virus RNA in clinical samples. Biosensors and Bioelectronics. 2017; 92: 349–356.
- Mohammadi J, Moattari A, Sattarahmady N, Pirbonyeh N, Yadegari H, Heli H. Electrochemical biosensing of influenza A subtype genome based on meso/macroporous cobalt (II) oxide nanoflakes-applied to human samples. Analytica Chimica Acta. 2017; 979: 51–57.
- Ilkhani H, Farhad S. A novel electrochemical DNA biosensor for Ebola virus detection. Analytical Biochemistry. 2018; 557: 151–155.
- Marrazza G, Ramalingam M, Jaisankar A, Cheng L, Selvolini G, Vitale IA. Advancements and emerging technologies in biosensors for rapid and accurate virus detection. TrAC Trends in Analytical Chemistry. 2024; 172: 117609.
|
- Rezaei B, Jamei HR, and Ensafi AA. An ultrasensitive and selective electrochemical aptasensor based on RGO-MWCNTs/Chitosan/carbon quantum dot for the detection of lysozyme. Biosensors and Bioelectronics. 2018; 115: 37-44.
- Izadi Z, Sheikh-Zeinoddin M, Ensafi AA, Soleimanian-Zad S. Fabrication of an electrochemical DNA-based biosensor for Bacillus cereus detection in milk and infant formula. Biosensors and Bioelectronics. 2016; 80: 582–589. https://doi.org/10.1016/j.bios.2016.02.032.
- Mansor NA, Zain ZM, Hamzah HH, Noorden MSA, Jaapar SS, Beni V, Ibupoto ZH. Detection of Breast Cancer 1 (BRCA1) Gene Using an Electrochemical DNA Biosensor Based on Immobilized ZnO Nanowires. Open Journal of Applied Biosensors. 2014; 3: 9–17.
- Izuan J, Rashid A, Azah N. The strategies of DNA immobilization and hybridization detection mechanism in the construction of electrochemical DNA sensor : A review. Sensing and Bio-Sensing Research. 2017; 16: 19–31.
- Chun L, Kim SE, Cho M, Cheo WS, Nam J, Lee DW, Lee Y. Electrochemical detection of HER2 using single stranded DNA aptamer modified gold nanoparticles electrode. Sensors and Actuators B. 2013; 186: 446–450.
- Kavita V J. DNA Biosensors-A Review. Journal of Bioengineering Biomedical Science. 2017; 7: 1-5. DOI: 10.4172/2155-9538.1000222.
- Gunasekaran B M, Srinivasan S, Ezhilan M, Nesakumar N. Nucleic acid-based electrochemical biosensors. Clinica Chimica Acta. 2024; 559: 119715. https://doi.org/10.1016/j.cca.2024.119715.
- Eksin E, Yildirim A, Bozoglu A, Zor E, Erdem, A. Paper-based nucleic acid biosensors. TrAC Trends in Analytical Chemistry. 2024; 171: 117511. https://doi.org/10.1016/j.trac.2023.117511.