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      Metagenomics and the molecular identification of novel viruses

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          Abstract

          There have been rapid recent developments in establishing methods for identifying and characterising viruses associated with animal and human diseases. These methodologies, commonly based on hybridisation or PCR techniques, are combined with advanced sequencing techniques termed ‘next generation sequencing’. Allied advances in data analysis, including the use of computational transcriptome subtraction, have also impacted the field of viral pathogen discovery. This review details these molecular detection techniques, discusses their application in viral discovery, and provides an overview of some of the novel viruses discovered. The problems encountered in attributing disease causality to a newly identified virus are also considered.

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          Cloning of a human parvovirus by molecular screening of respiratory tract samples.

          The identification of new virus species is a key issue for the study of infectious disease but is technically very difficult. We developed a system for large-scale molecular virus screening of clinical samples based on host DNA depletion, random PCR amplification, large-scale sequencing, and bioinformatics. The technology was applied to pooled human respiratory tract samples. The first experiments detected seven human virus species without the use of any specific reagent. Among the detected viruses were one coronavirus and one parvovirus, both of which were at that time uncharacterized. The parvovirus, provisionally named human bocavirus, was in a retrospective clinical study detected in 17 additional patients and associated with lower respiratory tract infections in children. The molecular virus screening procedure provides a general culture-independent solution to the problem of detecting unknown virus species in single or pooled samples. We suggest that a systematic exploration of the viruses that infect humans, "the human virome," can be initiated.
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            Identification of herpesvirus-like DNA sequences in AIDS-associated Kaposi's sarcoma.

            Representational difference analysis was used to isolate unique sequences present in more than 90 percent of Kaposi's sarcoma (KS) tissues obtained from patients with acquired immunodeficiency syndrome (AIDS). These sequences were not present in tissue DNA from non-AIDS patients, but were present in 15 percent of non-KS tissue DNA samples from AIDS patients. The sequences are homologous to, but distinct from, capsid and tegument protein genes of the Gammaherpesvirinae, herpesvirus saimiri and Epstein-Barr virus. These KS-associated herpesvirus-like (KSHV) sequences appear to define a new human herpesvirus.
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              Sequencing technologies - the next generation.

              Demand has never been greater for revolutionary technologies that deliver fast, inexpensive and accurate genome information. This challenge has catalysed the development of next-generation sequencing (NGS) technologies. The inexpensive production of large volumes of sequence data is the primary advantage over conventional methods. Here, I present a technical review of template preparation, sequencing and imaging, genome alignment and assembly approaches, and recent advances in current and near-term commercially available NGS instruments. I also outline the broad range of applications for NGS technologies, in addition to providing guidelines for platform selection to address biological questions of interest.
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                Author and article information

                Contributors
                Journal
                Vet J
                Vet. J
                Veterinary Journal (London, England : 1997)
                Elsevier Ltd.
                1090-0233
                1532-2971
                15 December 2010
                November 2011
                15 December 2010
                : 190
                : 2
                : 191-198
                Affiliations
                [a ]Department of Veterinary Medicine, University of Cambridge, Cambridge CB3 0ES, UK
                [b ]The Wellcome Trust Sanger Institute, Hinxton, Cambridge CB10 1SA, UK
                Author notes
                [* ]Corresponding author. Tel.: +44 1223 765631; fax: +44 1223 330886. nb289@ 123456cam.ac.uk
                Article
                S1090-0233(10)00361-8
                10.1016/j.tvjl.2010.10.014
                7110547
                21111643
                4accdc24-43aa-45e9-9ec9-cfc664c7b4fa
                Copyright © 2010 Elsevier Ltd. All rights reserved.

                Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.

                History
                : 20 October 2010
                Categories
                Article

                Veterinary medicine
                metagenomics,virus discovery,animals,computational transcriptome subtraction,hybridisation

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