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

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Ilié, Marius | Benzaquen, Jonathan | Hofman, Véronique | Long-Mira, Elodie | Lassalle, Sandra | Boutros, Jacques | Bontoux, Christophe | Lespinet-Fabre, Virginie | Bordone, Olivier | Tanga, Virginie | Allegra, Maryline | Salah, Myriam | Fayada, Julien | Leroy, Sylvie | Vassallo, Matteo | Touitou, Irit | Courjon, Johan | Contenti, Julie | Carles, Michel | Marquette, Charles-Hugo | Hofman, Paul

Edité par CCSD ; MDPI -

International audience. 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.

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