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Targeted Whole Genome Sequencing of the Capripoxvirus Genome from Clinical Tissue Samples and Lyophilized Vaccine Batches.

Diergezondheid  
[1]
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Public Access

Published

Peer reviewed scientific article

Engels

DOI : https://doi.org/10.1007/978-1-0716-3515-5_12 [2]

Auteurs

Elisabeth Mathijs [3]; Frank Vandenbussche [4]

Trefwoorden

    Samenvatting:

    Diseases caused by Capripoxviruses (CaPVs) are of great economic importance in sheep, goats, and cattle. Since CaPV strains are serologically indistinguishable and genetically highly homologous, typing of closely related strains can only be achieved by whole-genome sequencing. In this chapter, we describe a robust, cost-effective, and widely applicable protocol for reconstructing (nearly) complete CaPV genomes directly from clinical samples or commercial vaccine batches in less than a week. Taking advantage of the genetic similarity of CaPVs, a set of pan-CaPVs long-range PCRs was developed…
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    Samenvatting

    Diseases caused by Capripoxviruses (CaPVs) are of great economic importance in sheep, goats, and cattle. Since CaPV strains are serologically indistinguishable and genetically highly homologous, typing of closely related strains can only be achieved by whole-genome sequencing. In this chapter, we describe a robust, cost-effective, and widely applicable protocol for reconstructing (nearly) complete CaPV genomes directly from clinical samples or commercial vaccine batches in less than a week. Taking advantage of the genetic similarity of CaPVs, a set of pan-CaPVs long-range PCRs was developed that covers the entire genome with only a limited number of tiled amplicons. The resulting amplicons can be sequenced on all currently available high-throughput sequencing platforms. As an example, we have included a detailed protocol for performing nanopore sequencing and a pipeline for assembling the resulting tiled amplicon data.

    Associated health topics:

    Diergezondheid [5]

    Source URL:https://sciensano.be/nl/biblio/targeted-whole-genome-sequencing-capripoxvirus-genome-clinical-tissue-samples-and-lyophilized

    Links
    [1] https://sciensano.be/sites/default/files/viral_metagenomics.pdf [2] https://doi.org/10.1007/978-1-0716-3515-5_12 [3] https://sciensano.be/nl/people/elisabeth-mathijs/biblio [4] https://sciensano.be/nl/people/frank-vandenbussche/biblio [5] https://sciensano.be/nl/gezondheidsonderwerpen/diergezondheid