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Sci Rep ; 10(1): 17492, 2020 10 15.
Article in English | MEDLINE | ID: covidwho-872738

ABSTRACT

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.


Subject(s)
Betacoronavirus/genetics , Genome, Viral , Nanopore Sequencing/methods , Adolescent , Adult , Betacoronavirus/classification , Betacoronavirus/isolation & purification , COVID-19 , Child , Coronavirus Infections/diagnosis , Coronavirus Infections/virology , Female , Genetic Variation , Humans , Male , Middle Aged , Mutation, Missense , Pandemics , Phylogeny , Pneumonia, Viral/diagnosis , Pneumonia, Viral/virology , SARS-CoV-2 , Sequence Analysis, DNA , Young Adult
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