Field-Adapted Full Genome Sequencing of Peste-Des-Petits-Ruminants Virus Using Nanopore Sequencing

Peste-des-petits-ruminants virus (PPRV) is currently the focus of a control and eradication program. Full genome sequencing has the opportunity to become a powerful tool in the eradication program by improving molecular epidemiology and the study of viral evolution. PPRV is prevalent in many resourc...

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Autores principales: Torsson, Emeli, Kgotlele, Tebogo, Misinzo, Gerald, Johansson Wensman, Jonas, Berg, Mikael, Karlsson Lindsjö, Oskar
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2020
Materias:
Veterinary Science
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7649132/
https://www.ncbi.nlm.nih.gov/pubmed/33195515
http://dx.doi.org/10.3389/fvets.2020.542724
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author Torsson, Emeli
Kgotlele, Tebogo
Misinzo, Gerald
Johansson Wensman, Jonas
Berg, Mikael
Karlsson Lindsjö, Oskar
author_facet Torsson, Emeli
Kgotlele, Tebogo
Misinzo, Gerald
Johansson Wensman, Jonas
Berg, Mikael
Karlsson Lindsjö, Oskar
author_sort Torsson, Emeli
collection PubMed
description Peste-des-petits-ruminants virus (PPRV) is currently the focus of a control and eradication program. Full genome sequencing has the opportunity to become a powerful tool in the eradication program by improving molecular epidemiology and the study of viral evolution. PPRV is prevalent in many resource-constrained areas, with long distances to laboratory facilities, which can lack the correct equipment for high-throughput sequencing. Here we present a protocol for near full or full genome sequencing of PPRV. The use of a portable miniPCR and MinION brings the laboratory to the field and in addition makes the production of a full genome possible within 24 h of sampling. The protocol has been successfully used on virus isolates from cell cultures and field isolates from tissue samples of naturally infected goats.
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spelling pubmed-76491322020-11-13 Field-Adapted Full Genome Sequencing of Peste-Des-Petits-Ruminants Virus Using Nanopore Sequencing Torsson, Emeli Kgotlele, Tebogo Misinzo, Gerald Johansson Wensman, Jonas Berg, Mikael Karlsson Lindsjö, Oskar Front Vet Sci Veterinary Science Peste-des-petits-ruminants virus (PPRV) is currently the focus of a control and eradication program. Full genome sequencing has the opportunity to become a powerful tool in the eradication program by improving molecular epidemiology and the study of viral evolution. PPRV is prevalent in many resource-constrained areas, with long distances to laboratory facilities, which can lack the correct equipment for high-throughput sequencing. Here we present a protocol for near full or full genome sequencing of PPRV. The use of a portable miniPCR and MinION brings the laboratory to the field and in addition makes the production of a full genome possible within 24 h of sampling. The protocol has been successfully used on virus isolates from cell cultures and field isolates from tissue samples of naturally infected goats. Frontiers Media S.A. 2020-10-26 /pmc/articles/PMC7649132/ /pubmed/33195515 http://dx.doi.org/10.3389/fvets.2020.542724 Text en Copyright © 2020 Torsson, Kgotlele, Misinzo, Johansson Wensman, Berg and Karlsson Lindsjö. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Veterinary Science
Torsson, Emeli
Kgotlele, Tebogo
Misinzo, Gerald
Johansson Wensman, Jonas
Berg, Mikael
Karlsson Lindsjö, Oskar
Field-Adapted Full Genome Sequencing of Peste-Des-Petits-Ruminants Virus Using Nanopore Sequencing
title Field-Adapted Full Genome Sequencing of Peste-Des-Petits-Ruminants Virus Using Nanopore Sequencing
title_full Field-Adapted Full Genome Sequencing of Peste-Des-Petits-Ruminants Virus Using Nanopore Sequencing
title_fullStr Field-Adapted Full Genome Sequencing of Peste-Des-Petits-Ruminants Virus Using Nanopore Sequencing
title_full_unstemmed Field-Adapted Full Genome Sequencing of Peste-Des-Petits-Ruminants Virus Using Nanopore Sequencing
title_short Field-Adapted Full Genome Sequencing of Peste-Des-Petits-Ruminants Virus Using Nanopore Sequencing
title_sort field-adapted full genome sequencing of peste-des-petits-ruminants virus using nanopore sequencing
topic Veterinary Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7649132/
https://www.ncbi.nlm.nih.gov/pubmed/33195515
http://dx.doi.org/10.3389/fvets.2020.542724
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