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High-coverage SARS-CoV-2 genome sequences acquired by target capture sequencing

Authors :
Pan-Xin Du
Shao-Qing Wen
Chang Sun
Zhe Liu
Lirong Zou
Huanying Zheng
Xuding Xu
Ling-Xiang Wang
Xiaofang Peng
Guangyi Zeng
Jie Wu
Bo Lei
Fang Chen
Huan Zhang
Xiao Zhang
Changwen Ke
Wei Zhang
Jiyuan Yang
Lijun Liang
Source :
Journal of Medical Virology
Publication Year :
2020

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 120nt ssDNA 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 a 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 (GVM). 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. This article is protected by copyright. All rights reserved.

Details

ISSN :
10969071
Volume :
92
Issue :
10
Database :
OpenAIRE
Journal :
Journal of medical virology
Accession number :
edsair.doi.dedup.....d75ac8da6d55a0f9ad8f86c4a70529e3