tetano
Editor, Senior Moderator
Open Forum Infect Dis
. 2021 Jul 14;8(8)
fab364.
doi: 10.1093/ofid/ofab364. eCollection 2021 Aug.
Rapid Base-Specific Calling of SARS-CoV-2 Variants of Concern Using Combined RT-PCR Melting Curve Screening and SIRPH Technology
Sascha Tierling[SUP] 1 [/SUP], Kathrin Kattler[SUP] 1 [/SUP], Markus Vogelgesang[SUP] 2 [/SUP], Thorsten Pfuhl[SUP] 2 [/SUP], Stefan Lohse[SUP] 2 [/SUP], Christina Lo Porto[SUP] 1 [/SUP], Beate Schmitt[SUP] 1 [/SUP], Seiwert Nastasja[SUP] 2 [/SUP], Abdulrahman Salhab[SUP] 1 [/SUP], Sigrun Smola[SUP] 2 [/SUP], Jörn Walter[SUP] 1 [/SUP]
Affiliations
Abstract
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.
Keywords: SARS-CoV-2; SIRPH; SNuPE; mutation screening; variants of concern.
. 2021 Jul 14;8(8)
fab364.doi: 10.1093/ofid/ofab364. eCollection 2021 Aug.
Rapid Base-Specific Calling of SARS-CoV-2 Variants of Concern Using Combined RT-PCR Melting Curve Screening and SIRPH Technology
Sascha Tierling[SUP] 1 [/SUP], Kathrin Kattler[SUP] 1 [/SUP], Markus Vogelgesang[SUP] 2 [/SUP], Thorsten Pfuhl[SUP] 2 [/SUP], Stefan Lohse[SUP] 2 [/SUP], Christina Lo Porto[SUP] 1 [/SUP], Beate Schmitt[SUP] 1 [/SUP], Seiwert Nastasja[SUP] 2 [/SUP], Abdulrahman Salhab[SUP] 1 [/SUP], Sigrun Smola[SUP] 2 [/SUP], Jörn Walter[SUP] 1 [/SUP]
Affiliations
- PMID: 34409122
- PMCID: PMC8364759
- DOI: 10.1093/ofid/ofab364
Abstract
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.
Keywords: SARS-CoV-2; SIRPH; SNuPE; mutation screening; variants of concern.