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Rapid genomic characterization of SARS-CoV-2 viruses from clinical specimens using nanopore sequencing

The novel SARS-CoV-2 outbreak has swiftly spread worldwide. The rapid genome sequencing of SARS-CoV-2 strains has become a helpful tool for better understanding the genomic characteristics and origin of the virus. To obtain virus whole-genome sequences directly from clinical specimens, we performed...

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Autores principales: Li, Jun, Wang, Haoqiu, Mao, Lingfeng, Yu, Hua, Yu, Xinfen, Sun, Zhou, Qian, Xin, Cheng, Shi, Chen, Shuchang, Chen, Junfang, Pan, Jingcao, Shi, Jueliang, Wang, Xuchu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7566627/
https://www.ncbi.nlm.nih.gov/pubmed/33060796
http://dx.doi.org/10.1038/s41598-020-74656-y
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author Li, Jun
Wang, Haoqiu
Mao, Lingfeng
Yu, Hua
Yu, Xinfen
Sun, Zhou
Qian, Xin
Cheng, Shi
Chen, Shuchang
Chen, Junfang
Pan, Jingcao
Shi, Jueliang
Wang, Xuchu
author_facet Li, Jun
Wang, Haoqiu
Mao, Lingfeng
Yu, Hua
Yu, Xinfen
Sun, Zhou
Qian, Xin
Cheng, Shi
Chen, Shuchang
Chen, Junfang
Pan, Jingcao
Shi, Jueliang
Wang, Xuchu
author_sort Li, Jun
collection PubMed
description The novel SARS-CoV-2 outbreak has swiftly spread worldwide. The rapid genome sequencing of SARS-CoV-2 strains has become a helpful tool for better understanding the genomic characteristics and origin of the virus. To obtain virus whole-genome sequences directly from clinical specimens, we performed nanopore sequencing using a modified ARTIC protocol in a portable nanopore sequencer and validated a routine 8-h workflow and a 5-h rapid pipeline. We conducted some optimization to improve the genome sequencing workflow. The sensitivity of the workflow was also tested by serially diluting RNA from clinical samples. The optimized pipeline was finally applied to obtain the whole genomes of 29 clinical specimens collected in Hangzhou from January to March 2020. In the 29 obtained complete genomes of SARS-CoV-2, 33 variations were identified and analyzed. The genomic variations and phylogenetic analysis hinted at multiple sources and different transmission patterns during the COVID-19 epidemic in Hangzhou, China. In conclusion, the genomic characteristics and origin of the virus can be quickly determined by nanopore sequencing following our workflows.
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spelling pubmed-75666272020-10-19 Rapid genomic characterization of SARS-CoV-2 viruses from clinical specimens using nanopore sequencing Li, Jun Wang, Haoqiu Mao, Lingfeng Yu, Hua Yu, Xinfen Sun, Zhou Qian, Xin Cheng, Shi Chen, Shuchang Chen, Junfang Pan, Jingcao Shi, Jueliang Wang, Xuchu Sci Rep Article The novel SARS-CoV-2 outbreak has swiftly spread worldwide. The rapid genome sequencing of SARS-CoV-2 strains has become a helpful tool for better understanding the genomic characteristics and origin of the virus. To obtain virus whole-genome sequences directly from clinical specimens, we performed nanopore sequencing using a modified ARTIC protocol in a portable nanopore sequencer and validated a routine 8-h workflow and a 5-h rapid pipeline. We conducted some optimization to improve the genome sequencing workflow. The sensitivity of the workflow was also tested by serially diluting RNA from clinical samples. The optimized pipeline was finally applied to obtain the whole genomes of 29 clinical specimens collected in Hangzhou from January to March 2020. In the 29 obtained complete genomes of SARS-CoV-2, 33 variations were identified and analyzed. The genomic variations and phylogenetic analysis hinted at multiple sources and different transmission patterns during the COVID-19 epidemic in Hangzhou, China. In conclusion, the genomic characteristics and origin of the virus can be quickly determined by nanopore sequencing following our workflows. Nature Publishing Group UK 2020-10-15 /pmc/articles/PMC7566627/ /pubmed/33060796 http://dx.doi.org/10.1038/s41598-020-74656-y Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Li, Jun
Wang, Haoqiu
Mao, Lingfeng
Yu, Hua
Yu, Xinfen
Sun, Zhou
Qian, Xin
Cheng, Shi
Chen, Shuchang
Chen, Junfang
Pan, Jingcao
Shi, Jueliang
Wang, Xuchu
Rapid genomic characterization of SARS-CoV-2 viruses from clinical specimens using nanopore sequencing
title Rapid genomic characterization of SARS-CoV-2 viruses from clinical specimens using nanopore sequencing
title_full Rapid genomic characterization of SARS-CoV-2 viruses from clinical specimens using nanopore sequencing
title_fullStr Rapid genomic characterization of SARS-CoV-2 viruses from clinical specimens using nanopore sequencing
title_full_unstemmed Rapid genomic characterization of SARS-CoV-2 viruses from clinical specimens using nanopore sequencing
title_short Rapid genomic characterization of SARS-CoV-2 viruses from clinical specimens using nanopore sequencing
title_sort rapid genomic characterization of sars-cov-2 viruses from clinical specimens using nanopore sequencing
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7566627/
https://www.ncbi.nlm.nih.gov/pubmed/33060796
http://dx.doi.org/10.1038/s41598-020-74656-y
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