Giuseppe
Emeritus
Development of reverse transcription-loop-mediated isothermal amplification assay for pandemic (H1N1) 2009 virus as a novel molecular based testing for pandemic influenza even in resource limited settings. (J Clin Microbiol., abstract, RA-1018, edited)
J Clin Microbiol. 2010 Jan 13. [Epub ahead of print]
Development of reverse transcription-loop-mediated isothermal amplification assay for pandemic (H1N1) 2009 virus as a novel molecular based testing for pandemic influenza even in resource limited settings.
Kubo T, Agoh M, Mai LQ, Fukushima K, Nishimura H, Yamaguchi A, Hirano M, Yoshikawa A, Hasebe F, Kohno S, Morita K. - Center for International Collaborative Research, Institute for Tropical Medicine, Nagasaki University, 1-12-4, Sakamoto, Nagasaki, 852-8523, Japan; Department of Virology, Institute of Tropical Medicine, Nagasaki University, 1-12-4, Sakamoto, Nagasaki, 852-8523, Japan; Nagasaki Prefectural Institute for Environmental Research and Public Health, 2-1306-11, Ikeda, Omura, Nagasaki, Japan; National Institute of Hygiene and Epidemiology, No1 Yersin Street, Hai Ba Trung District, Hanoi 10000, Vietnam; Division of Respiratory Medicine, Japanese Red Cross Nagasaki Genbaku Isahaya Hospital, 986-2, Keya, Tarami, Isahaya, Nagasaki, Japan; Virus Research Center, Clinical Research Division, Sendai Medical Center, National Hospital Organization, 2-8-8 Miyagino, Sendai, Miyagi, 983-8520, Japan; Department of Molecular Microbiology and Immunology, Nagasaki University Graduate School of Biomedical Sciences, 1-12-4, Sakamoto, Nagasaki, 852-8523, Japan.
This paper reports on the development of a one-step, real-time reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay targeting hemagglutitin (HA) gene for the rapid detection of pandemic (H1N1) 2009 virus. Detection limit of the pandemic (H1N1) 2009-HA-specific RT-LAMP was same as that of currently used real-time RT-PCR method. The assay detected pandemic (H1N1) 2009 HA gene in 136 RNA samples extracted from Japanese and Vietnamese patients' nasopharyngeal swab specimens. No cross-reactive amplification with other seasonal influenza virus RNA was observed and the detection of specific viral genome targets in clinical specimens was achieved in less than 40 minutes. The sensitivity and specificity of pandemic (H1N1) 2009-HA-specific RT-LAMP assay obtained in this study were 97.8% and 100%, respectively. Pandemic (H1N1) 2009-HA-specific RT-LAMP assay will enable faster and easier diagnoses of pandemic (H1N1) 2009 infection especially in the resource limited situations in developing countries. Here we propose a new method of rapid molecular based testing for pandemic influenza.
PMID: 20071551 [PubMed - as supplied by publisher]
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J Clin Microbiol. 2010 Jan 13. [Epub ahead of print]
Development of reverse transcription-loop-mediated isothermal amplification assay for pandemic (H1N1) 2009 virus as a novel molecular based testing for pandemic influenza even in resource limited settings.
Kubo T, Agoh M, Mai LQ, Fukushima K, Nishimura H, Yamaguchi A, Hirano M, Yoshikawa A, Hasebe F, Kohno S, Morita K. - Center for International Collaborative Research, Institute for Tropical Medicine, Nagasaki University, 1-12-4, Sakamoto, Nagasaki, 852-8523, Japan; Department of Virology, Institute of Tropical Medicine, Nagasaki University, 1-12-4, Sakamoto, Nagasaki, 852-8523, Japan; Nagasaki Prefectural Institute for Environmental Research and Public Health, 2-1306-11, Ikeda, Omura, Nagasaki, Japan; National Institute of Hygiene and Epidemiology, No1 Yersin Street, Hai Ba Trung District, Hanoi 10000, Vietnam; Division of Respiratory Medicine, Japanese Red Cross Nagasaki Genbaku Isahaya Hospital, 986-2, Keya, Tarami, Isahaya, Nagasaki, Japan; Virus Research Center, Clinical Research Division, Sendai Medical Center, National Hospital Organization, 2-8-8 Miyagino, Sendai, Miyagi, 983-8520, Japan; Department of Molecular Microbiology and Immunology, Nagasaki University Graduate School of Biomedical Sciences, 1-12-4, Sakamoto, Nagasaki, 852-8523, Japan.
This paper reports on the development of a one-step, real-time reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay targeting hemagglutitin (HA) gene for the rapid detection of pandemic (H1N1) 2009 virus. Detection limit of the pandemic (H1N1) 2009-HA-specific RT-LAMP was same as that of currently used real-time RT-PCR method. The assay detected pandemic (H1N1) 2009 HA gene in 136 RNA samples extracted from Japanese and Vietnamese patients' nasopharyngeal swab specimens. No cross-reactive amplification with other seasonal influenza virus RNA was observed and the detection of specific viral genome targets in clinical specimens was achieved in less than 40 minutes. The sensitivity and specificity of pandemic (H1N1) 2009-HA-specific RT-LAMP assay obtained in this study were 97.8% and 100%, respectively. Pandemic (H1N1) 2009-HA-specific RT-LAMP assay will enable faster and easier diagnoses of pandemic (H1N1) 2009 infection especially in the resource limited situations in developing countries. Here we propose a new method of rapid molecular based testing for pandemic influenza.
PMID: 20071551 [PubMed - as supplied by publisher]
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