Rapid Base-Specific Calling of SARS-CoV-2 Variants of Concern Using Combined RT-PCR Melting Curve Screening and SIRPH Technology-Reference-Cited by-同舟云学术

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

Published:2021-07-14 Issue:8 Volume:8 Page:
ISSN:2328-8957
Container-title:Open Forum Infectious Diseases
language:en
Short-container-title:

Author:

Tierling Sascha¹,Kattler Kathrin¹^ORCID,Vogelgesang Markus²,Pfuhl Thorsten²,Lohse Stefan²,Lo Porto Christina¹,Schmitt Beate¹,Nastasja Seiwert²,Salhab Abdulrahman¹,Smola Sigrun²,Walter Jörn¹^ORCID

Affiliation:

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

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

Abstract

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.

Funder

German Federal Ministry of Research and Education

Publisher

Oxford University Press (OUP)

Subject

Infectious Diseases,Oncology

Link

http://academic.oup.com/ofid/advance-article-pdf/doi/10.1093/ofid/ofab364/38949175/ofab364.pdf

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