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Article

Optimized qRT-PCR Approach for the Detection of Intra- and Extra-Cellular SARS-CoV-2 RNAs

1
Institute of Medical Virology, University Hospital Frankfurt am Main, Goethe University, 60590 Frankfurt am Main, Germany
2
GenXPro GmbH, Frankfurter Innovationszentrum, Biotechnologie (FIZ), 60438 Frankfurt am Main, Germany
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2020, 21(12), 4396; https://doi.org/10.3390/ijms21124396
Received: 18 April 2020 / Revised: 8 June 2020 / Accepted: 18 June 2020 / Published: 20 June 2020
(This article belongs to the Special Issue Molecular Detection and Typing of Viruses)
The novel coronavirus SARS-CoV-2 is the causative agent of the acute respiratory disease COVID-19, which has become a global concern due to its rapid spread. Meanwhile, increased demand for testing has led to a shortage of reagents and supplies and compromised the performance of diagnostic laboratories in many countries. Both the World Health Organization (WHO) and the Center for Disease Control and Prevention (CDC) recommend multi-step RT-PCR assays using multiple primer and probe pairs, which might complicate the interpretation of the test results, especially for borderline cases. In this study, we describe an alternative RT-PCR approach for the detection of SARS-CoV-2 RNA that can be used for the probe-based detection of clinical isolates in diagnostics as well as in research labs using a low-cost SYBR green method. For the evaluation, we used samples from patients with confirmed SARS-CoV-2 infections and performed RT-PCR assays along with successive dilutions of RNA standards to determine the limit of detection. We identified an M-gene binding primer and probe pair highly suitable for the quantitative detection of SARS-CoV-2 RNA for diagnostic and research purposes. View Full-Text
Keywords: SARS-CoV-2; COVID-19; qRT-PCR detection; test protocol; coronavirus SARS-CoV-2; COVID-19; qRT-PCR detection; test protocol; coronavirus
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MDPI and ACS Style

Toptan, T.; Hoehl, S.; Westhaus, S.; Bojkova, D.; Berger, A.; Rotter, B.; Hoffmeier, K.; Cinatl, J., Jr.; Ciesek, S.; Widera, M. Optimized qRT-PCR Approach for the Detection of Intra- and Extra-Cellular SARS-CoV-2 RNAs. Int. J. Mol. Sci. 2020, 21, 4396. https://doi.org/10.3390/ijms21124396

AMA Style

Toptan T, Hoehl S, Westhaus S, Bojkova D, Berger A, Rotter B, Hoffmeier K, Cinatl J Jr., Ciesek S, Widera M. Optimized qRT-PCR Approach for the Detection of Intra- and Extra-Cellular SARS-CoV-2 RNAs. International Journal of Molecular Sciences. 2020; 21(12):4396. https://doi.org/10.3390/ijms21124396

Chicago/Turabian Style

Toptan, Tuna, Sebastian Hoehl, Sandra Westhaus, Denisa Bojkova, Annemarie Berger, Björn Rotter, Klaus Hoffmeier, Jindrich Cinatl Jr., Sandra Ciesek, and Marek Widera. 2020. "Optimized qRT-PCR Approach for the Detection of Intra- and Extra-Cellular SARS-CoV-2 RNAs" International Journal of Molecular Sciences 21, no. 12: 4396. https://doi.org/10.3390/ijms21124396

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