Genosensor for rapid,sensitive, specific point-of-care detection of H1N1 influenza (swine flu) |
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| Institution: | 1. Centre for Medical Biotechnology, Maharshi Dayanand University, Rohtak, Haryana, 124007, India;2. DCR University of Science and Technology, Murthal, Sonipat, 131027, India;3. Pt.BhagwatDyal Sharma Post Graduate Institute of Medical Sciences, Rohtak, Haryana 124001, India;4. Department of Biotechnology, Jamia Hamdard, Delhi, India;5. Department of Animal Biotechnology, Lala Lajpat Rai University of Veterinary & Animal Sciences, Hisar, 125004, Haryana India;1. Department of Analytical Chemistry, Institute of Chemistry, State University of São Paulo (UNESP), 14801-970, Araraquara, SP, Brazil;2. Grupo de Eletroquímica Aplicada a Polímeros e Sensores, Instituto de Química, Universidade Federal de Uberlândia, 38700-128, Patos de Minas, MG, Brazil;3. Oswaldo Cruz-Fiocruz Foundation, Aggeu Magalhães Institute, Av. Prof. Moraes Rego s/nº, Cidade Universitária, 50670-420, P.O. Box 7.472, Recife, PE, Brazil;4. Biomedical Engineering Laboratory, Federal University of Pernambuco, Av. Prof. Moraes Rego, 1235, Cidade Universitaria, 50670-901, Recife, PE, Brazil;1. Department of Bioscience and Biotechnology, Konkuk University, Gwangjin-gu, Seoul, 05029, South Korea;2. Department of Emergency Medicine, Kyung Hee University Hospital at Gangdong, Seoul, 05278, South Korea;3. Department of Pediatrics, Korea Cancer Center Hospital, Nowon-gu, Seoul, 01812, South Korea;1. Polish Academy of Sciences, The Szewalski Institute of Fluid-Flow Machinery, 14 Fiszera St., 80-231, Gdańsk, Poland;2. Department of Analytical Chemistry, Faculty of Chemistry, University of Gdańsk, 63 Wita Stwosza St., 80-308, Gdańsk, Poland;3. Department of Metrology and Optoelectronics, Faculty of Electronics, Telecommunications and Informatics, Gdańsk University of Technology, 11/12 G. Narutowicza St., 80-233, Gdańsk, Poland;4. Institute of Biotechnology and Molecular Medicine, 3 Trzy Lipy St., 80-172, Gdańsk, Poland;5. SensDx, 14b Postępu St., 02-676, Warszawa, Poland;1. Luminescent MD, LLC, Hagerstown, MD 21742, United States;2. Virginia Polytech Inst & State Univ, Dept Wood Sci & Froest Prod, Blacksburg, VA 24061, United States;1. Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran;2. Zabol University of Medical Sciences, Zabol, Iran;3. Drag Applied Research Center, Tabriz University of Medical Sciences, Tabriz 51664, Iran;4. Department of Biotechnology, Higher Education Institute of Rab-Rashid, Tabriz, Iran;5. Department of Analytical Chemistry, University of Valencia, Dr. Moliner 50, 46100 Burjassot, Valencia, Spain |
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| Abstract: | A 5′ amine group-linked haemagglutinin (HA) gene-specific probe was attached over the surface of a working electrode to develop a rapid, specific, and sensitive point of care detection assay for H1N1 (swine flu) in human respiratory nasal swabs. The probe was attached with a cysteine covered screen-printed gold electrode via 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide (EDC/NHS). The electrochemical assay was performed using differential pulse voltammetry with the use of the redox indicator methylene blue for the detection of different concentrations of the single-stranded viral genome. The developed genosensor showed high sensitivity for H1N1 influenza virus with a detection limit of 0.002 ng/6 μL of viral nucleic acid in the sample. Samples were analysed by quantitative real-time Polymerase Chain Reaction as well as by conventional PCR. The genosensor showed high specificity, as no cross-reaction was observed with the heterologous nucleic acid of different pathogens (Salmonella typhi, Neisseria meningitides, and Streptococcus pyogenes) and human DNA, and it was specific for H1N1 with a sensitivity of ∼49 μA cm−2 ng-1. Genosensor is based on a very simple methodology that can be followed based on its easy-to-access approach. It is quick and could be used as a point-of-care test for the detection of influenza virus within 30 min. |
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| Keywords: | Biosensor DPV EDC-NHS H1N1 RNA Genosensor POC test |
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