Accurate Detection of SARS-CoV-2 by Next-Generation Sequencing in Low Viral Load Specimens

Int J Mol Sci. 2023 Feb 9;24(4):3478. doi: 10.3390/ijms24043478.

Abstract

As new SARS-CoV-2 variants emerge, there is an urgent need to increase the efficiency and availability of viral genome sequencing, notably to detect the lineage in samples with a low viral load. SARS-CoV-2 genome next-generation sequencing (NGS) was performed retrospectively in a single center on 175 positive samples from individuals. An automated workflow used the Ion AmpliSeq SARS-CoV-2 Insight Research Assay on the Genexus Sequencer. All samples were collected in the metropolitan area of the city of Nice (France) over a period of 32 weeks (from 19 July 2021 to 11 February 2022). In total, 76% of cases were identified with a low viral load (Ct ≥ 32, and ≤200 copies/µL). The NGS analysis was successful in 91% of cases, among which 57% of cases harbored the Delta variant, and 34% the Omicron BA.1.1 variant. Only 9% of cases had unreadable sequences. There was no significant difference in the viral load in patients infected with the Omicron variant compared to the Delta variant (Ct values, p = 0.0507; copy number, p = 0.252). We show that the NGS analysis of the SARS-CoV-2 genome provides reliable detection of the Delta and Omicron SARS-CoV-2 variants in low viral load samples.

Keywords: COVID-19; Genexus; SARS-CoV-2; low viral load; next generation sequencing.

MeSH terms

  • COVID-19*
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Retrospective Studies
  • SARS-CoV-2*
  • Viral Load

Supplementary concepts

  • SARS-CoV-2 variants

Grants and funding

This work was supported by Agence Régionale de Santé Provence-Alpes-Côte d’Azur, Conseil Départemental 06 des Alpes Maritimes, Ville de Nice, Métropole Nice Côte d’Azur, and Fonds de Dotation AVENI.