tetano
Editor, Senior Moderator
iScience
. 2021 Jul 21;102892.
doi: 10.1016/j.isci.2021.102892. Online ahead of print.
A benchmarking study of SARS-CoV-2 whole-genome sequencing protocols using COVID-19 patient samples
Tiantian Liu[SUP] 1 [/SUP], Zhong Chen[SUP] 1 [/SUP], Wanqiu Chen[SUP] 1 [/SUP], Xin Chen[SUP] 1 2 [/SUP], Maryam Hosseini[SUP] 1 [/SUP], Zhaowei Yang[SUP] 3 1 [/SUP], Jing Li[SUP] 3 2 [/SUP], Diana Ho[SUP] 1 [/SUP], David Turay[SUP] 4 [/SUP], Ciprian P Gheorghe[SUP] 5 [/SUP], Wendell Jones[SUP] 6 [/SUP], Charles Wang[SUP] 1 2 [/SUP]
Affiliations
Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an emerging new type of coronavirus that is responsible for the COVID-19 pandemic and the unprecedented global health emergency. Whole-genome sequencing (WGS) of SARS-CoV-2 plays a critical role in understanding the disease. Performance variation exists across SARS-CoV-2 viral WGS technologies, but there is currently no benchmarking study comparing different WGS sequencing protocols. We compared seven different SARS-CoV-2 WGS library protocols using RNA from patient nasopharyngeal swab samples under two storage conditions with low and high viral inputs. We found large differences in mappability and genome coverage, and variations in sensitivity, reproducibility and precision of single-nucleotide variant calling across different protocols. For certain amplicon-based protocol, appropriate primer trimming step is critical for accurate single-nucleotide variant calling. We ranked the performance of protocols based on six different metrics. Our findings offer guidance in choosing appropriate WGS protocols to characterize SARS-CoV-2 and its evolution.
. 2021 Jul 21;102892.
doi: 10.1016/j.isci.2021.102892. Online ahead of print.
A benchmarking study of SARS-CoV-2 whole-genome sequencing protocols using COVID-19 patient samples
Tiantian Liu[SUP] 1 [/SUP], Zhong Chen[SUP] 1 [/SUP], Wanqiu Chen[SUP] 1 [/SUP], Xin Chen[SUP] 1 2 [/SUP], Maryam Hosseini[SUP] 1 [/SUP], Zhaowei Yang[SUP] 3 1 [/SUP], Jing Li[SUP] 3 2 [/SUP], Diana Ho[SUP] 1 [/SUP], David Turay[SUP] 4 [/SUP], Ciprian P Gheorghe[SUP] 5 [/SUP], Wendell Jones[SUP] 6 [/SUP], Charles Wang[SUP] 1 2 [/SUP]
Affiliations
- PMID: 34308277
- PMCID: PMC8294598
- DOI: 10.1016/j.isci.2021.102892
Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an emerging new type of coronavirus that is responsible for the COVID-19 pandemic and the unprecedented global health emergency. Whole-genome sequencing (WGS) of SARS-CoV-2 plays a critical role in understanding the disease. Performance variation exists across SARS-CoV-2 viral WGS technologies, but there is currently no benchmarking study comparing different WGS sequencing protocols. We compared seven different SARS-CoV-2 WGS library protocols using RNA from patient nasopharyngeal swab samples under two storage conditions with low and high viral inputs. We found large differences in mappability and genome coverage, and variations in sensitivity, reproducibility and precision of single-nucleotide variant calling across different protocols. For certain amplicon-based protocol, appropriate primer trimming step is critical for accurate single-nucleotide variant calling. We ranked the performance of protocols based on six different metrics. Our findings offer guidance in choosing appropriate WGS protocols to characterize SARS-CoV-2 and its evolution.