- 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.
MetadatenAuthor: | Tuna ToptanORCiDGND, Sebastian HoehlORCiDGND, Sandra WesthausGND, Denisa BojkovaORCiDGND, Annemarie BergerGND, Björn RotterORCiD, Klaus Hoffmeier, Jindrich CinatlORCiDGND, Sandra CiesekORCiDGND, Marek WideraORCiDGND |
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URN: | urn:nbn:de:hebis:30:3-547710 |
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DOI: | https://doi.org/10.3390/ijms21124396 |
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ISSN: | 1422-0067 |
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ISSN: | 1661-6596 |
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Pubmed Id: | https://pubmed.ncbi.nlm.nih.gov/32575728 |
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Parent Title (English): | International journal of molecular sciences |
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Publisher: | Molecular Diversity Preservation International |
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Place of publication: | Basel |
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Document Type: | Article |
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Language: | English |
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Year of Completion: | 2020 |
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Date of first Publication: | 2020/06/20 |
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Publishing Institution: | Universitätsbibliothek Johann Christian Senckenberg |
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Release Date: | 2020/07/06 |
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Tag: | COVID-19; SARS-CoV-2; coronavirus; qRT-PCR detection; test protocol |
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Volume: | 21 |
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Issue: | 20, Art. 4396 |
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Page Number: | 11 |
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First Page: | 1 |
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Last Page: | 11 |
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Note: | This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited |
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Note: | Preprint erschienen in: bioRxiv beta, 2020, doi:10.1101/2020.04.20.052258 |
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HeBIS-PPN: | 467515476 |
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Institutes: | Medizin / Medizin |
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Dewey Decimal Classification: | 6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit |
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Sammlungen: | Universitätspublikationen |
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Open-Access-Publikationsfonds: | Medizin |
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Licence (German): | Creative Commons - Namensnennung 4.0 |
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