There has been a need for rapid detection of Avian Influenza virus (AIV) H5N1 due to it being a potential pandemic threat. Most of the current methods, including culture isolation and PCR, are very sensitive and specific but require specialized laboratories and trained personnel in order to complete the tests and are time-consuming. The goal of this study was to design a biosensor that would be able to rapidly detect AIV H5N1 using aptamers as biosensing material and a quartz crystal microbalance (QCM) for transducing method. Specific DNA aptamers against AIV H5N1 were immobilized, through biotin and streptavidin conjugation, onto the gold surface of QCM sensor to capture the target virus. Magnetic nanobeads (150 nm in diameter) were then added as amplifiers considering its large surface/volume ratio which allows for faster movement and a higher target molecule binding rate. The result showed that the captured AIV caused frequency change, and more change was observed when the AIV concentration increased. The nanobead amplification was effective at the lower concentrations of AIV, however, it was not significant when the AIV concentration was 1 HA or higher. The detection limit of the aptasensor was 1 HAU with a detection time of 1 h. The capture of the target virus on to the surface of QCM sensor and the binding of magnetic nanobeads with the virus was confirmed with electron microscopy. Aptamers have unlimited shelf life and are temperature stable which allows this aptasensor to give much more consistent results specifically for in field applications.
A. Burns, D. van der Mensbrugghe and H. Timmer, “Evaluating the Economic Consequences of Avian Influenza,” World Bank Report, 2008.
B. Charlton, B. Crossley and S. Hietala, “Conventional and Future Diagnostics for Avian Influenza,” Comparative Immunology Microbiology and Infectious Diseases, Vol. 32, No. 4, 2009, pp. 341-350.
Y. Amano and Q. Cheng, “Detection of Influenza Virus: Traditional Approaches and Development of Biosensors,” Analytical and Bioanalytical Chemistry, Vol. 381, No. 1, 2005, pp. 156-164.
C. Estmer-Nilsson, S. Abbas, M. Bennemo, A. Larsson, M. D. H?m?l?inen and ?. Frostell-Karlsson, “A Novel Assay for Influenza Virus Quantification Using Surface Plasmon Resonance,” Vaccine, Vol. 28, No. 3, 2010, pp. 759-766. http://dx.doi.org/10.1016/j.vaccine.2009.10.070
J. Xu, D. Suarez and D. S. Gottfried, “Detection of Avian Influenza Virus Using an Interferometric Biosensor,” Analytical and Bioanalytical Chemistry, Vol. 389, No. 1, 2007, pp. 1193-1199.
R. Wang, Y. Wang, K. Lassiter, Y. Li, B. Hargis, S. Tung, et al., “Interdigitated Array Microelectrode Based Impedance Immunosensor for the Detection of Avian Influenza Virus HN1,” Talanta, Vol. 79, No. 2, 2009, pp. 159- 164. http://dx.doi.org/10.1016/j.talanta.2009.03.017
J. Lum, R. Wang, K. Lassiter, B. Srinivasan, D. Abi-Ghanem, L. Berghman, et al., “Rapid Detection of Avian In- fluenza H5N1 Virus Using Impedance Measurement of Immuno-Reaction Coupled with RBC Amplification,” Biosensors and Bioelectronics, Vol. 38, 2012, pp. 67-73.
D. Ivnitski, I. Abdel-Hamid, P. Atanasov and E. Wilkins, “Biosensors for Detection of Pathogenic Bacteria,” Bio- sensors and Bioelectronics, Vol. 14, 1999, pp. 599-624.
F. Liu, Y. Li, X. Su, M. F. Slavik, Y. Ying and J. Wang, “QCM Immunosensor with Nanoparticle Amplification for Detection of Escherichia coli O157:H7,” Sensing and Instrumentation for Food Quality, Vol. 1, 2007, pp. 161-168. http://dx.doi.org/10.1007/s11694-007-9021-1
T. M. P. Hewa, G. A. Tannock, D. E. Mainwaring, S. Harrison and J. V. Fecondo, “The Detection of Influenza A and B Viruses in Clinical Specimens Using a Quartz Crystal Microbalance,” Journal of Virological Methods, Vol. 162, No. 1-2, 2009, pp. 14-21.
D. Li, J. Wang, R. Wang, Y. Li, D. Abi-Ghanem, L. Berghman, B. Hargis and H. Lu, “A Nanobeads Amplified QCM Immunosensor for the Detection of Avian Influenza Virus H5N1,” Biosensors and Bioelectronics, Vol. 26, 2011, pp. 4146-4154.
T. W. Owen, R. O. Al-Kaysi, C. J. Bardeen and Q. Cheng, “Microgravimetric Immunosensor for Direct Detection of Aerosolized Influenza A Virus Particles,” Sensors and Actuators B, Vol. 126, 2007, pp. 691-699.
C. Tuerk and L. Gold, “Systematic Evolution of Ligands by Exponential Enrichment: RNA Ligands to Bacteriophage T4 DNA Polymerase,” Science, Vol. 249, No. 4968, 1990, pp. 505-510.
K. Sefah, J. A. Phillips, X. Xiong, L. Meng, D. Van Simaeys, H. Chen, et al., “Nucleic Acid Aptamers for Biosensors and Bio-Analytical Applications,” Analyst, Vol. 134, 2009, pp. 1765-1775.
Z. Cui, Q. Ren, H. Wei, Z. Chen, J. Deng, Z. Zhang, et al., “Quantum Dot-Aptamer Nanoprobes for Recognizing and Labeling Influenza A Virus Particles,” Nanoscale, Vol. 3, 2011, pp. 2454-2457.