Rapid Base-Specific Calling of SARS-CoV-2 Variants of Concern Using Combined RT-PCR Melting Curve Screening and SIRPH Technology.

SARS-CoV-2 SIRPH SNuPE mutation screening variants of concern

Journal

Open forum infectious diseases
ISSN: 2328-8957
Titre abrégé: Open Forum Infect Dis
Pays: United States
ID NLM: 101637045

Informations de publication

Date de publication:
Aug 2021
Historique:
received: 14 04 2021
accepted: 12 07 2021
entrez: 19 8 2021
pubmed: 20 8 2021
medline: 20 8 2021
Statut: epublish

Résumé

The emergence of novel variants of concern of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) demands fast and reliable detection of such variants in local populations. Here we present a cost-efficient and fast workflow combining a prescreening of SARS-CoV-2-positive samples using reverse transcription polymerase chain reaction melting curve analysis with multiplexed IP-RP-HPLC-based single nucleotide primer extensions. The entire workflow from positive SARS-CoV-2 testing to base-specific identification of variants requires about 24 hours. We applied the sensitive method to monitor local variant of concern outbreaks in SARS-CoV-2-positive samples collected in a confined region of Germany.

Sections du résumé

BACKGROUND BACKGROUND
The emergence of novel variants of concern of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) demands fast and reliable detection of such variants in local populations.
METHODS METHODS
Here we present a cost-efficient and fast workflow combining a prescreening of SARS-CoV-2-positive samples using reverse transcription polymerase chain reaction melting curve analysis with multiplexed IP-RP-HPLC-based single nucleotide primer extensions.
RESULTS RESULTS
The entire workflow from positive SARS-CoV-2 testing to base-specific identification of variants requires about 24 hours.
CONCLUSIONS CONCLUSIONS
We applied the sensitive method to monitor local variant of concern outbreaks in SARS-CoV-2-positive samples collected in a confined region of Germany.

Identifiants

pubmed: 34409122
doi: 10.1093/ofid/ofab364
pii: ofab364
pmc: PMC8364759
doi:

Types de publication

Journal Article

Langues

eng

Pagination

ofab364

Informations de copyright

© The Author(s) 2021. Published by Oxford University Press on behalf of Infectious Diseases Society of America.

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Auteurs

Sascha Tierling (S)

Department of Genetics, Saarland University, Saarbrücken, Saarland, Germany.

Kathrin Kattler (K)

Department of Genetics, Saarland University, Saarbrücken, Saarland, Germany.

Markus Vogelgesang (M)

Institute of Virology, Saarland University Medical Center, Homburg, Saarland, Germany.

Thorsten Pfuhl (T)

Institute of Virology, Saarland University Medical Center, Homburg, Saarland, Germany.

Stefan Lohse (S)

Institute of Virology, Saarland University Medical Center, Homburg, Saarland, Germany.

Christina Lo Porto (C)

Department of Genetics, Saarland University, Saarbrücken, Saarland, Germany.

Beate Schmitt (B)

Department of Genetics, Saarland University, Saarbrücken, Saarland, Germany.

Seiwert Nastasja (S)

Institute of Virology, Saarland University Medical Center, Homburg, Saarland, Germany.

Abdulrahman Salhab (A)

Department of Genetics, Saarland University, Saarbrücken, Saarland, Germany.

Sigrun Smola (S)

Institute of Virology, Saarland University Medical Center, Homburg, Saarland, Germany.

Jörn Walter (J)

Department of Genetics, Saarland University, Saarbrücken, Saarland, Germany.

Classifications MeSH