tetano
Editor, Senior Moderator
PLoS One
. 2024 Apr 29;19(4):e0289188.
doi: 10.1371/journal.pone.0289188. eCollection 2024. Performance of amplicon and capture based next-generation sequencing approaches for the epidemiological surveillance of Omicron SARS-CoV-2 and other variants of concern
Carlos Daviña-Núñez[SUP] 1 2 [/SUP], Sonia Pérez[SUP] 1 3 [/SUP], Jorge Julio Cabrera-Alvargonzález[SUP] 1 3 [/SUP], Anniris Rincón-Quintero[SUP] 1 3 [/SUP], Ana Treinta-Álvarez[SUP] 3 [/SUP], Montse Godoy-Diz[SUP] 3 [/SUP], Silvia Suárez-Luque[SUP] 4 [/SUP], Benito Regueiro-García[SUP] 1 [/SUP]
Affiliations
To control the SARS-CoV-2 pandemic, healthcare systems have focused on ramping up their capacity for epidemiological surveillance through viral whole genome sequencing. In this paper, we tested the performance of two protocols of SARS-CoV-2 nucleic acid enrichment, an amplicon enrichment using different versions of the ARTIC primer panel and a hybrid-capture method using KAPA RNA Hypercap. We focused on the challenge of the Omicron variant sequencing, the advantages of automated library preparation and the influence of the bioinformatic analysis in the final consensus sequence. All 94 samples were sequenced using Illumina iSeq 100 and analysed with two bioinformatic pipelines: a custom-made pipeline and an Illumina-owned pipeline. We were unsuccessful in sequencing six samples using the capture enrichment due to low reads. On the other hand, amplicon dropout and mispriming caused the loss of mutation G21987A and the erroneous addition of mutation T15521A respectively using amplicon enrichment. Overall, we found high sequence agreement regardless of method of enrichment, bioinformatic pipeline or the use of automation for library preparation in eight different SARS-CoV-2 variants. Automation and the use of a simple app for bioinformatic analysis can simplify the genotyping process, making it available for more diagnostic facilities and increasing global vigilance.
. 2024 Apr 29;19(4):e0289188.
doi: 10.1371/journal.pone.0289188. eCollection 2024. Performance of amplicon and capture based next-generation sequencing approaches for the epidemiological surveillance of Omicron SARS-CoV-2 and other variants of concern
Carlos Daviña-Núñez[SUP] 1 2 [/SUP], Sonia Pérez[SUP] 1 3 [/SUP], Jorge Julio Cabrera-Alvargonzález[SUP] 1 3 [/SUP], Anniris Rincón-Quintero[SUP] 1 3 [/SUP], Ana Treinta-Álvarez[SUP] 3 [/SUP], Montse Godoy-Diz[SUP] 3 [/SUP], Silvia Suárez-Luque[SUP] 4 [/SUP], Benito Regueiro-García[SUP] 1 [/SUP]
Affiliations
- PMID: 38683803
- DOI: 10.1371/journal.pone.0289188
To control the SARS-CoV-2 pandemic, healthcare systems have focused on ramping up their capacity for epidemiological surveillance through viral whole genome sequencing. In this paper, we tested the performance of two protocols of SARS-CoV-2 nucleic acid enrichment, an amplicon enrichment using different versions of the ARTIC primer panel and a hybrid-capture method using KAPA RNA Hypercap. We focused on the challenge of the Omicron variant sequencing, the advantages of automated library preparation and the influence of the bioinformatic analysis in the final consensus sequence. All 94 samples were sequenced using Illumina iSeq 100 and analysed with two bioinformatic pipelines: a custom-made pipeline and an Illumina-owned pipeline. We were unsuccessful in sequencing six samples using the capture enrichment due to low reads. On the other hand, amplicon dropout and mispriming caused the loss of mutation G21987A and the erroneous addition of mutation T15521A respectively using amplicon enrichment. Overall, we found high sequence agreement regardless of method of enrichment, bioinformatic pipeline or the use of automation for library preparation in eight different SARS-CoV-2 variants. Automation and the use of a simple app for bioinformatic analysis can simplify the genotyping process, making it available for more diagnostic facilities and increasing global vigilance.