Development of a simple genotyping method based on indel mutations to rapidly screen SARS-CoV-2 circulating variants: Delta, Omicron BA.1 and BA.2.


Journal

Journal of virological methods
ISSN: 1879-0984
Titre abrégé: J Virol Methods
Pays: Netherlands
ID NLM: 8005839

Informations de publication

Date de publication:
09 2022
Historique:
received: 15 04 2022
revised: 13 06 2022
accepted: 16 06 2022
pubmed: 21 6 2022
medline: 19 7 2022
entrez: 20 6 2022
Statut: ppublish

Résumé

The high need of rapid and flexible tools that facilitate the identification of circulating SARS-CoV-2 Variants of Concern (VOCs) remains crucial for public health system monitoring. Here, we develop allele-specific (AS)-qPCR assays targeting three recurrent indel mutations, ΔEF156-157, Ins214EPE and ΔLPP24-26, in spike (S) gene to identify the Delta VOC and the Omicron sublineages BA.1 and BA.2, respectively. After verification of the analytical specificity of each primer set, two duplex qPCR assays with melting curve analysis were performed to screen 129 COVID-19 cases confirmed between December 31, 2021 and February 01, 2022 in Sfax, Tunisia. The first duplex assay targeting ΔEF156-157 and Ins214EPE mutations successfully detected the Delta VOC in 39 cases and Omicron BA.1 in 83 cases. All the remaining cases (n = 7) were identified as Omicron BA.2, by the second duplex assay targeting Ins214EPE and ΔLPP24-26 mutations. The results of the screening method were in perfect concordance with those of S gene partial sequencing. In conclusion, our findings provide a simple and flexible screening method for more rapid and reliable monitoring of circulating VOCs. We highly recommend its implementation to guide public health policies.

Identifiants

pubmed: 35724698
pii: S0166-0934(22)00117-3
doi: 10.1016/j.jviromet.2022.114570
pmc: PMC9212420
pii:
doi:

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

114570

Informations de copyright

Copyright © 2022 Elsevier B.V. All rights reserved.

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Auteurs

Wajdi Ayadi (W)

Laboratory of Molecular Biotechnology of Eukaryotes, Center of Biotechnology of Sfax, University of Sfax, Sidi Mansour Street Km 6, BP 1177, 3038 Sfax, Tunisia. Electronic address: wajdi.ayadi.cbs@gmail.com.

Awatef Taktak (A)

Laboratory of Molecular Biotechnology of Eukaryotes, Center of Biotechnology of Sfax, University of Sfax, Sidi Mansour Street Km 6, BP 1177, 3038 Sfax, Tunisia; Laboratory of Microbiology, Habib Bourguiba University-Hospital, Sfax, Tunisia.

Saba Gargouri (S)

Laboratory of Microbiology, Habib Bourguiba University-Hospital, Sfax, Tunisia; Faculty of Medicine, University of Sfax, Sfax, Tunisia.

Fahmi Smaoui (F)

Laboratory of Microbiology, Habib Bourguiba University-Hospital, Sfax, Tunisia.

Amel Chtourou (A)

Laboratory of Microbiology, Habib Bourguiba University-Hospital, Sfax, Tunisia; Faculty of Medicine, University of Sfax, Sfax, Tunisia.

Houda Skouri-Gargouri (H)

Laboratory of Molecular Biotechnology of Eukaryotes, Center of Biotechnology of Sfax, University of Sfax, Sidi Mansour Street Km 6, BP 1177, 3038 Sfax, Tunisia.

Rihab Derbel (R)

Laboratory of Microbiology, Habib Bourguiba University-Hospital, Sfax, Tunisia.

Azza Hadj Sassi (AH)

Laboratory of Molecular Biotechnology of Eukaryotes, Center of Biotechnology of Sfax, University of Sfax, Sidi Mansour Street Km 6, BP 1177, 3038 Sfax, Tunisia.

Ali Gargouri (A)

Laboratory of Molecular Biotechnology of Eukaryotes, Center of Biotechnology of Sfax, University of Sfax, Sidi Mansour Street Km 6, BP 1177, 3038 Sfax, Tunisia.

Adnene Hammami (A)

Laboratory of Microbiology, Habib Bourguiba University-Hospital, Sfax, Tunisia; Faculty of Medicine, University of Sfax, Sfax, Tunisia.

Héla Karray-Hakim (H)

Laboratory of Microbiology, Habib Bourguiba University-Hospital, Sfax, Tunisia; Faculty of Medicine, University of Sfax, Sfax, Tunisia.

Raja Mokdad-Gargouri (R)

Laboratory of Molecular Biotechnology of Eukaryotes, Center of Biotechnology of Sfax, University of Sfax, Sidi Mansour Street Km 6, BP 1177, 3038 Sfax, Tunisia.

Lamia Fki-Berrajah (L)

Laboratory of Microbiology, Habib Bourguiba University-Hospital, Sfax, Tunisia; Faculty of Medicine, University of Sfax, Sfax, Tunisia.

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