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J Med Virol ; 92(10): 2221-2226, 2020 10.
Article in English | MEDLINE | ID: covidwho-505569

ABSTRACT

In this study, we designed a set of SARS-CoV-2 enrichment probes to increase the capacity for sequence-based virus detection and obtain the comprehensive genome sequence at the same time. This universal SARS-CoV-2 enrichment probe set contains 502 120 nt single-stranded DNA biotin-labeled probes designed based on all available SARS-CoV-2 viral sequences and it can be used to enrich for SARS-CoV-2 sequences without prior knowledge of type or subtype. Following the CDC health and safety guidelines, marked enrichment was demonstrated in a virus strain sample from cell culture, three nasopharyngeal swab samples (cycle threshold [Ct ] values: 32.36, 36.72, and 38.44) from patients diagnosed with COVID-19 (positive control) and four throat swab samples from patients without COVID-19 (negative controls), respectively. Moreover, based on these high-quality sequences, we discuss the heterozygosity and viral expression during coronavirus replication and its phylogenetic relationship with other selected high-quality samples from the Genome Variation Map. Therefore, this universal SARS-CoV-2 enrichment probe system can capture and enrich SARS-CoV-2 viral sequences selectively and effectively in different samples, especially clinical swab samples with a relatively low concentration of viral particles.


Subject(s)
COVID-19/diagnosis , DNA Probes/metabolism , DNA, Single-Stranded/genetics , Genome, Viral , SARS-CoV-2/genetics , Whole Genome Sequencing/methods , Biotin/chemistry , COVID-19/pathology , COVID-19/virology , DNA Probes/chemical synthesis , DNA, Single-Stranded/metabolism , Genotype , Humans , Mutation , Nasopharynx/virology , Phylogeny , Reverse Transcriptase Polymerase Chain Reaction/standards , SARS-CoV-2/classification , SARS-CoV-2/isolation & purification , SARS-CoV-2/pathogenicity , Sensitivity and Specificity
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