Direct detection of unamplified hepatitis C virus RNA using unmodified gold nanoparticles |
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| Authors: | Sherif M. Shawky Dirk Bald Hassan M.E. Azzazy |
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| Affiliation: | 1. Department of Chemistry & Yousef Jameel Science & Technology Research Center, the American University in Cairo, P.O. Box 74 New Cairo 11835, Egypt;2. Department of Structural Biology, Faculty of Earth & Life Science, Free University, Amsterdam, The Netherlands;1. Institute of Nano Electronic Engineering, Universiti Malaysia Perlis, 01000 Kangar, Perlis, Malaysia;2. School of Microelectronic Engineering, Universiti Malaysia Perlis, 01000 Kangar, Perlis, Malaysia;3. School of Bioprocess Engineering, Universiti Malaysia Perlis, 02600 Arau, Perlis, Malaysia;2. School of Basic Medical Science, Jiangxi University of Chinese Traditional Medicine, Nanchang, Jiangxi, 330000, China;3. Food and Nutritional Science, School of Biological Sciences, The University of Hong Kong, Pokfulam Road, Hong Kong;1. School of Biomedical Engineering, Sun Yat-sen University, Shenzhen, 518107, China;2. School of Life Sciences, Jilin University, Changchun, 130012, China;3. Engineering Research Center of Chinese Ministry of Education for Edible and Medicinal Fungi, Jilin Agricultural University, Changchun, 130118, China;4. Shanghai Institute of Immunology, Department of Immunology and Microbiology, Shanghai Jiao Tong University School of Medicine, Shanghai, China;5. Institute of Environmental Research at Greater Bay Area, Key Laboratory for Water Quality and Conservation of the Pearl River Delta, Ministry of Education, Guangzhou University, Guangzhou, 510006, China;6. Laboratory of Biomaterials and Translational Medicine, Center for Nanomedicine, The Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, 510630, China;7. Research Center of Translational Medicine, Shanghai Children’s Hospital, Shanghai, 200025, China;8. Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China;1. CENTEX Shrimp, Faculty of Science, Mahidol University, Rama 6 Road, Bangkok 10400, Thailand;2. National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Pathumthani 12120, Thailand;1. School of Ocean Sciences, Bangor University, Menai Bridge, Anglesey, UK;2. School of Natural Sciences, Bangor University, Deiniol Road, Bangor, Gwynedd, UK;3. Centre for Environment, Fisheries and Aquaculture Science, Weymouth, Dorset, UK |
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| Abstract: | BackgroundGold nanoparticles (AuNPs) exhibit a unique phenomenon known as Surface Plasmon Resonance, which is responsible for their intense red color. This color changes to blue upon aggregation of AuNPs.ObjectiveThis work aims to develop a rapid, simple and cheap assay for direct detection of unamplified HCV RNA extracted from clinical samples using unmodified AuNPs.MethodsSerum samples were collected from healthy volunteers (n = 45) and chronic HCV patients (n = 30). Extracted RNA, hybridization buffer containing PBS, and a primer targeting the 5′UTR of HCV were mixed. The mixture was denatured, annealed, and then cooled to room temperature for 10 min followed by addition of AuNPs.ResultsSalt, primer, AuNPs concentrations and annealing temperature and time were all optimized. In HCV positive specimens, the color of the solution changed from red to blue within 1 min. The assay has a sensitivity of 92%, a specificity of 88.9%, and a detection limit of 50 copies/reaction.ConclusionsTo our knowledge, this is the first assay that allows the detection of unamplified HCV RNA in clinical specimens using unmodified AuNPs. The developed assay is highly sensitive, has a turnaround time of 30 min, and eliminates the need for thermal cycling and detection instruments. |
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