tetano
Editor, Senior Moderator
Pathogens
. 2024 Jan 9;13(1):61.
doi: 10.3390/pathogens13010061. NASCarD (Nanopore Adaptive Sampling with Carrier DNA): A Rapid, PCR-Free Method for SARS-CoV-2 Whole-Genome Sequencing in Clinical Samples
Miguel A Terrazos Miani[SUP] 1 [/SUP], Loïc Borcard[SUP] 1 [/SUP], Sonja Gempeler[SUP] 1 [/SUP], Christian Baumann[SUP] 1 [/SUP], Pascal Bittel[SUP] 1 [/SUP], Stephen L Leib[SUP] 1 [/SUP], Stefan Neuenschwander[SUP] 1 [/SUP], Alban Ramette[SUP] 1 [/SUP]
Affiliations
Whole-genome sequencing (WGS) represents the main technology for SARS-CoV-2 lineage characterization in diagnostic laboratories worldwide. The rapid, near-full-length sequencing of the viral genome is commonly enabled by high-throughput sequencing of PCR amplicons derived from cDNA molecules. Here, we present a new approach called NASCarD (Nanopore Adaptive Sampling with Carrier DNA), which allows a low amount of nucleic acids to be sequenced while selectively enriching for sequences of interest, hence limiting the production of non-target sequences. Using COVID-19 positive samples available during the omicron wave, we demonstrate how the method may lead to >99% genome completeness of the SARS-CoV-2 genome sequences within 7 h of sequencing at a competitive cost. The new approach may have applications beyond SARS-CoV-2 sequencing for other DNA or RNA pathogens in clinical samples.
Keywords: SARS-CoV-2; beta coronaviruses; genomics; nanopore sequencing; next-generation sequencing; whole-genome sequencing.
. 2024 Jan 9;13(1):61.
doi: 10.3390/pathogens13010061. NASCarD (Nanopore Adaptive Sampling with Carrier DNA): A Rapid, PCR-Free Method for SARS-CoV-2 Whole-Genome Sequencing in Clinical Samples
Miguel A Terrazos Miani[SUP] 1 [/SUP], Loïc Borcard[SUP] 1 [/SUP], Sonja Gempeler[SUP] 1 [/SUP], Christian Baumann[SUP] 1 [/SUP], Pascal Bittel[SUP] 1 [/SUP], Stephen L Leib[SUP] 1 [/SUP], Stefan Neuenschwander[SUP] 1 [/SUP], Alban Ramette[SUP] 1 [/SUP]
Affiliations
- PMID: 38251368
- DOI: 10.3390/pathogens13010061
Whole-genome sequencing (WGS) represents the main technology for SARS-CoV-2 lineage characterization in diagnostic laboratories worldwide. The rapid, near-full-length sequencing of the viral genome is commonly enabled by high-throughput sequencing of PCR amplicons derived from cDNA molecules. Here, we present a new approach called NASCarD (Nanopore Adaptive Sampling with Carrier DNA), which allows a low amount of nucleic acids to be sequenced while selectively enriching for sequences of interest, hence limiting the production of non-target sequences. Using COVID-19 positive samples available during the omicron wave, we demonstrate how the method may lead to >99% genome completeness of the SARS-CoV-2 genome sequences within 7 h of sequencing at a competitive cost. The new approach may have applications beyond SARS-CoV-2 sequencing for other DNA or RNA pathogens in clinical samples.
Keywords: SARS-CoV-2; beta coronaviruses; genomics; nanopore sequencing; next-generation sequencing; whole-genome sequencing.