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      Discovery of a subgenotype of human coronavirus NL63 associated with severe lower respiratory tract infection in China, 2018

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          ABSTRACT

          Human coronavirus NL63 (HCoV-NL63) is primarily associated with common cold in children, elderly and immunocompromised individuals. Outbreaks caused by HCoV-NL63 are rare. Here we report a cluster of HCoV-NL63 cases with severe lower respiratory tract infection that arose in Guangzhou, China, in 2018. Twenty-three hospitalized children were confirmed to be HCoV-NL63 positive, and most of whom were hospitalized with severe pneumonia or acute bronchitis. Whole genomes of HCoV-NL63 were obtained using next-generation sequencing. Phylogenetic and single amino acid polymorphism analyses showed that this outbreak was associated with two subgenotypes (C3 and B) of HCoV-NL63. Half of patients were identified to be related to a new subgenotype C3. One unique amino acid mutation at I507 L in spike protein receptor binding domain (RBD) was detected, which segregated this subgenotype C3 from other known subgenotypes. Pseudotyped virus bearing the I507 L mutation in RBD showed enhanced entry into host cells as compared to the prototype virus. This study proved that HCoV-NL63 was undergoing continuous mutation and has the potential to cause severe lower respiratory disease in humans.

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          Isolation of a novel coronavirus from a man with pneumonia in Saudi Arabia.

          A previously unknown coronavirus was isolated from the sputum of a 60-year-old man who presented with acute pneumonia and subsequent renal failure with a fatal outcome in Saudi Arabia. The virus (called HCoV-EMC) replicated readily in cell culture, producing cytopathic effects of rounding, detachment, and syncytium formation. The virus represents a novel betacoronavirus species. The closest known relatives are bat coronaviruses HKU4 and HKU5. Here, the clinical data, virus isolation, and molecular identification are presented. The clinical picture was remarkably similar to that of the severe acute respiratory syndrome (SARS) outbreak in 2003 and reminds us that animal coronaviruses can cause severe disease in humans.
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            Characterization and complete genome sequence of a novel coronavirus, coronavirus HKU1, from patients with pneumonia.

            Despite extensive laboratory investigations in patients with respiratory tract infections, no microbiological cause can be identified in a significant proportion of patients. In the past 3 years, several novel respiratory viruses, including human metapneumovirus, severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV), and human coronavirus NL63, were discovered. Here we report the discovery of another novel coronavirus, coronavirus HKU1 (CoV-HKU1), from a 71-year-old man with pneumonia who had just returned from Shenzhen, China. Quantitative reverse transcription-PCR showed that the amount of CoV-HKU1 RNA was 8.5 to 9.6 x 10(6) copies per ml in his nasopharyngeal aspirates (NPAs) during the first week of the illness and dropped progressively to undetectable levels in subsequent weeks. He developed increasing serum levels of specific antibodies against the recombinant nucleocapsid protein of CoV-HKU1, with immunoglobulin M (IgM) titers of 1:20, 1:40, and 1:80 and IgG titers of <1:1,000, 1:2,000, and 1:8,000 in the first, second and fourth weeks of the illness, respectively. Isolation of the virus by using various cell lines, mixed neuron-glia culture, and intracerebral inoculation of suckling mice was unsuccessful. The complete genome sequence of CoV-HKU1 is a 29,926-nucleotide, polyadenylated RNA, with G+C content of 32%, the lowest among all known coronaviruses with available genome sequence. Phylogenetic analysis reveals that CoV-HKU1 is a new group 2 coronavirus. Screening of 400 NPAs, negative for SARS-CoV, from patients with respiratory illness during the SARS period identified the presence of CoV-HKU1 RNA in an additional specimen, with a viral load of 1.13 x 10(6) copies per ml, from a 35-year-old woman with pneumonia. Our data support the existence of a novel group 2 coronavirus associated with pneumonia in humans.
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              Full-length human immunodeficiency virus type 1 genomes from subtype C-infected seroconverters in India, with evidence of intersubtype recombination.

              The development of an effective human immunodeficiency virus type 1 (HIV-1) vaccine is likely to depend on knowledge of circulating variants of genes other than the commonly sequenced gag and env genes. In addition, full-genome data are particularly limited for HIV-1 subtype C, currently the most commonly transmitted subtype in India and worldwide. Likewise, little is known about sequence variation of HIV-1 in India, the country facing the largest burden of HIV worldwide. Therefore, the objective of this study was to clone and characterize the complete genome of HIV-1 from seroconverters infected with subtype C variants in India. Cocultured HIV-1 isolates were obtained from six seroincident individuals from Pune, India, and virtually full-length HIV-1 genomes were amplified, cloned, and sequenced from each. Sequence analysis revealed that five of the six genomes were of subtype C, while one was a mosaic of subtypes A and C, with multiple breakpoints in env, nef, and the 3' long terminal repeat as determined by both maximal chi2 analysis and phylogenetic bootstrapping. Sequences were compared for preservation of known cytotoxic T lymphocyte (CTL) epitopes. Compared with those of the HIV-1LAI sequence, 38% of well-defined CTL epitopes were identical. The proportion of nonconservative substitutions for Env, at 61%, was higher (P < 0.001) than those for Gag (24%), Pol (18%), and Nef (32%). Therefore, characterized CTL epitopes demonstrated substantial differences from subtype B laboratory strains, which were most pronounced in Env. Because these clones were obtained from Indian seroconverters, they are likely to facilitate vaccine-related efforts in India by providing potential antigens for vaccine candidates as well as for assays of vaccine responsiveness.
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                Author and article information

                Journal
                Emerg Microbes Infect
                Emerg Microbes Infect
                TEMI
                temi20
                Emerging Microbes & Infections
                Taylor & Francis
                2222-1751
                2020
                29 January 2020
                : 9
                : 1
                : 246-255
                Affiliations
                [a ]State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, the First Affiliated Hospital of Guangzhou Medical University , Guangzhou, People’s Republic of China
                [b ]Institute of Infectious disease, Guangzhou Eighth People's Hospital of Guangzhou Medical University , Guangzhou, People’s Republic of China
                [c ]The HKU–Pasteur Research Pole, School of Public Health, Li Ka Shing Faculty of Medicine, The University of Hong Kong , Hong Kong, People’s Republic of China
                Author notes
                [CONTACT ] Jincun Zhao zhaojincun@ 123456gird.cn State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, the First Affiliated Hospital of Guangzhou Medical University; Institute of Infectious disease, Guangzhou Eighth People's Hospital of Guangzhou Medical University , Guangzhou 510120, People’s Republic of China
                Rong Zhou zhourong@ 123456gird.cn State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, the First Affiliated Hospital of Guangzhou Medical University , Guangzhou 510120, People’s Republic of China
                Jingxian Zhao zhaojingxian@ 123456gird.cn State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, the First Affiliated Hospital of Guangzhou Medical University , Guangzhou 510120, People’s Republic of China
                [*]

                These authors contributed equally to this work.

                Supplemental data for this article can be accessed https://doi.org/10.1080/22221751.2020.1717999

                Article
                1717999
                10.1080/22221751.2020.1717999
                7034077
                31996093
                4b74d24e-8c75-4f7e-b65e-fc4b987da6e3
                © 2020 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group, on behalf of Shanghai Shangyixun Cultural Communication Co., Ltd

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 11 September 2019
                : 31 December 2019
                : 02 January 2020
                Page count
                Figures: 7, Tables: 0, Equations: 0, References: 33, Pages: 10
                Funding
                Funded by: National Key Research and Development Program of China
                Award ID: 2018YFC1200100 and 2016YFC1202701
                Funded by: National Science and Technology Major Project
                Award ID: 2018ZX10301403-001-003 and 2018ZX10101002
                Funded by: National Natural Science Foundation of China 10.13039/501100001809
                Award ID: 81702047
                Award ID: 81772191
                Award ID: 91842106 and 8181101118
                Funded by: Department of Science and Technology
                Funded by: State Key Laboratory of Respiratory Disease 10.13039/100013262
                Funded by: China Postdoctoral Science Foundation 10.13039/501100002858
                Funded by: Natural Science Foundation of Guangdong Province 10.13039/501100003453
                Funded by: Nanshan Medical Development Foundation of Guangdong Province, National Key Technology R&D Program
                Funded by: Guangdong Science and Technology Foundation 10.13039/501100007162
                This research was supported by grants from National Key Research and Development Program of China (2018YFC1200100 and 2016YFC1202701), National Science and Technology Major Project (2018ZX10301403-001-003 and 2018ZX10101002), National Natural Science Foundation of China (81702047, 81772191, 91842106 and 8181101118), Guangdong Provincial Department of Science and Technology (2018B020207013), the Thousand Talents Plan Award of China 2015 and State Key Laboratory of Respiratory Disease (SKLRD-QN-201715 and SKLRD-QN-201912), China Postdoctoral Science Foundation (2019T120722) and the PhD Start-up Fund of Natural Science Foundation of Guangdong Province, China, grants from the Nanshan Medical Development Foundation of Guangdong Province, National Key Technology R&D Program (2018YFC1311900), Guangdong Science and Technology Foundation (2019B030316028).
                Categories
                Article

                human coronavirus nl63,new subgenotype,pneumonia,whole-genome sequencing,phylogenetic analysis,viral entry

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