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      Complete Genomic Sequences of Highly Pathogenic H5N1 Avian Influenza Viruses Obtained Directly from Human Autopsy Specimens

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          Abstract

          The complete genomic sequences of H5N1 highly pathogenic avian influenza (HPAI) viruses were directly obtained from lung, trachea, and colon tissues and an intestinal fecal sample of a patient by using the next-generation sequencing technique. This is the first report on complete H5N1 viral genomes from human autopsy specimens.

          ABSTRACT

          The complete genomic sequences of H5N1 highly pathogenic avian influenza (HPAI) viruses were directly obtained from lung, trachea, and colon tissues and an intestinal fecal sample of a patient by using the next-generation sequencing technique. This is the first report on complete H5N1 viral genomes from human autopsy specimens.

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          Most cited references8

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          H5N1 infection of the respiratory tract and beyond: a molecular pathology study

          Summary Background Human infection with avian influenza H5N1 is an emerging infectious disease characterised by respiratory symptoms and a high fatality rate. Previous studies have shown that the human infection with avian influenza H5N1 could also target organs apart from the lungs. Methods We studied post-mortem tissues of two adults (one man and one pregnant woman) infected with H5N1 influenza virus, and a fetus carried by the woman. In-situ hybridisation (with sense and antisense probes to haemagglutinin and nucleoprotein) and immunohistochemistry (with monoclonal antibodies to haemagglutinin and nucleoprotein) were done on selected tissues. Reverse-transcriptase (RT) PCR, real-time RT-PCR, strand-specific RT-PCR, and nucleic acid sequence-based amplification (NASBA) detection assays were also undertaken to detect viral RNA in organ tissue samples. Findings We detected viral genomic sequences and antigens in type II epithelial cells of the lungs, ciliated and non-ciliated epithelial cells of the trachea, T cells of the lymph node, neurons of the brain, and Hofbauer cells and cytotrophoblasts of the placenta. Viral genomic sequences (but no viral antigens) were detected in the intestinal mucosa. In the fetus, we found viral sequences and antigens in the lungs, circulating mononuclear cells, and macrophages of the liver. The presence of viral sequences in the organs and the fetus was also confirmed by RT-PCR, strand-specific RT-PCR, real-time RT-PCR, and NASBA. Interpretation In addition to the lungs, H5N1 influenza virus infects the trachea and disseminates to other organs including the brain. The virus could also be transmitted from mother to fetus across the placenta.
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            Influenza A H5N1 Replication Sites in Humans

            Tissue tropism and pathogenesis of influenza A virus subtype H5N1 disease in humans is not well defined. In mammalian experimental models, H5N1 influenza is a disseminated disease. However, limited previous data from human autopsies have not shown evidence of virus dissemination beyond the lung. We investigated a patient with fatal H5N1 influenza. Viral RNA was detected by reverse transcription–polymerase chain reaction in lung, intestine, and spleen tissues, but positive-stranded viral RNA indicating virus replication was confined to the lung and intestine. Viral antigen was detected in pneumocytes by immunohistochemical tests. Tumor necrosis factor-α mRNA was seen in lung tissue. In contrast to disseminated infection documented in other mammals and birds, H5N1 viral replication in humans may be restricted to the lung and intestine, and the major site of H5N1 viral replication in the lung is the pneumocyte.
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              The gene structure and replication of influenza virus.

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                Author and article information

                Contributors
                Role: Editor
                Journal
                Microbiol Resour Announc
                Microbiol Resour Announc
                ga
                mra
                MRA
                Microbiology Resource Announcements
                American Society for Microbiology (1752 N St., N.W., Washington, DC )
                2576-098X
                6 December 2018
                December 2018
                : 7
                : 22
                : e01498-18
                Affiliations
                [a ]Department of Preventive and Social Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand
                [b ]Department of Pathology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand
                [c ]Department of Biochemistry, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand
                [d ]Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand
                [e ]Bureau of Epidemiology, Ministry of Public Health, Nonthaburi, Thailand
                [f ]Department of Pathology, Faculty of Medicine, Ramathibodi Hospital, Bangkok, Thailand
                [g ]Center for Research and Innovation, Faculty of Medical Technology, Mahidol University, Nakhon Pathom, Thailand
                Georgia Institute of Technology
                Author notes
                Address correspondence to Pilaipan Puthavathana, pilaipan.put@ 123456mahidol.edu .

                Citation Sangsiriwut K, Uiprasertkul M, Payungporn S, Auewarakul P, Ungchusak K, Chantratita W, Puthavathana P. 2018. Complete genomic sequences of highly pathogenic H5N1 avian influenza viruses obtained directly from human autopsy specimens. Microbiol Resour Announc 7:e01498-18. https://doi.org/10.1128/MRA.01498-18.

                Article
                MRA01498-18
                10.1128/MRA.01498-18
                6284082
                f620cbee-9b80-453a-ad48-2532922b749f
                Copyright © 2018 Sangsiriwut et al.

                This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

                History
                : 3 November 2018
                : 7 November 2018
                Page count
                Figures: 0, Tables: 1, Equations: 0, References: 9, Pages: 3, Words: 1462
                Funding
                Funded by: Mahidol University, https://doi.org/10.13039/501100004156;
                Award Recipient :
                Categories
                Genome Sequences
                Custom metadata
                December 2018

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