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

Tierling, Sascha; Kattler, Kathrin; Vogelgesang, Markus; Pfuhl, Thorsten; Lohse, Stefan; Lo Porto, Christina; Schmitt, Beate; Nastasja, Seiwert; Salhab, Abdulrahman; Smola, Sigrun; Walter, Jörn

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: 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: The entire workflow from positive SARS-CoV-2 testing to base-specific identification of variants requires about 24 hours. 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.