tetano
Editor, Senior Moderator
Sci Rep
. 2023 Apr 20;13(1):6461.
doi: 10.1038/s41598-023-33168-1.
Comparison of SARS-CoV-2 whole genome sequencing using tiled amplicon enrichment and bait hybridization
Anita Koskela von Sydow[SUP] 1 2 3 [/SUP], Carl Mårten Lindqvist[SUP] 4 5 [/SUP], Naveed Asghar[SUP] 4 [/SUP], Magnus Johansson[SUP] 4 [/SUP], Martin Sundqvist[SUP] 6 [/SUP], Paula Mölling[SUP] 5 6 [/SUP], Bianca Stenmark[SUP] 7 4 5 [/SUP]
Affiliations
Abstract
The severe acute respiratory syndrome coronavirus 2 (SARS‑CoV‑2) pandemic has led to extensive virological monitoring by whole genome sequencing (WGS). Investigating the advantages and limitations of different protocols is key when conducting population-level WGS. SARS-CoV-2 positive samples with Ct values of 14-30 were run using three different protocols: the Twist Bioscience SARS‑CoV‑2 protocol with bait hybridization enrichment sequenced with Illumina, and two tiled amplicon enrichment protocols, ARTIC V3 and Midnight, sequenced with Illumina and Oxford Nanopore Technologies, respectively. Twist resulted in better coverage uniformity and coverage of the entire genome, but has several drawbacks: high human contamination, laborious workflow, high cost, and variation between batches. The ARTIC and Midnight protocol produced an even coverage across samples, and almost all reads were mapped to the SARS-CoV-2 reference. ARTIC and Midnight represent robust, cost-effective, and highly scalable methods that are appropriate in a clinical environment. Lineage designations were uniform across methods, representing the dominant lineages in Sweden during the period of collection. This study provides insights into methodological differences in SARS‑CoV‑2 sequencing and guidance in selecting suitable methods for various purposes.
. 2023 Apr 20;13(1):6461.
doi: 10.1038/s41598-023-33168-1.
Comparison of SARS-CoV-2 whole genome sequencing using tiled amplicon enrichment and bait hybridization
Anita Koskela von Sydow[SUP] 1 2 3 [/SUP], Carl Mårten Lindqvist[SUP] 4 5 [/SUP], Naveed Asghar[SUP] 4 [/SUP], Magnus Johansson[SUP] 4 [/SUP], Martin Sundqvist[SUP] 6 [/SUP], Paula Mölling[SUP] 5 6 [/SUP], Bianca Stenmark[SUP] 7 4 5 [/SUP]
Affiliations
- PMID: 37081087
- DOI: 10.1038/s41598-023-33168-1
Abstract
The severe acute respiratory syndrome coronavirus 2 (SARS‑CoV‑2) pandemic has led to extensive virological monitoring by whole genome sequencing (WGS). Investigating the advantages and limitations of different protocols is key when conducting population-level WGS. SARS-CoV-2 positive samples with Ct values of 14-30 were run using three different protocols: the Twist Bioscience SARS‑CoV‑2 protocol with bait hybridization enrichment sequenced with Illumina, and two tiled amplicon enrichment protocols, ARTIC V3 and Midnight, sequenced with Illumina and Oxford Nanopore Technologies, respectively. Twist resulted in better coverage uniformity and coverage of the entire genome, but has several drawbacks: high human contamination, laborious workflow, high cost, and variation between batches. The ARTIC and Midnight protocol produced an even coverage across samples, and almost all reads were mapped to the SARS-CoV-2 reference. ARTIC and Midnight represent robust, cost-effective, and highly scalable methods that are appropriate in a clinical environment. Lineage designations were uniform across methods, representing the dominant lineages in Sweden during the period of collection. This study provides insights into methodological differences in SARS‑CoV‑2 sequencing and guidance in selecting suitable methods for various purposes.