Fast SARS-CoV-2 detection by RT-qPCR in preheated nasopharyngeal swab samples
Objectives: The gold-standard COVID-19 diagnosis relies on detecting SARS-CoV-2 using RNA purification and one-step retrotranscription and quantitative PCR (RT-qPCR). Based on the urgent need for high-throughput screening, we tested the performance of three alternative, simple and affordable protocols to rapidly detect SARS-CoV-2, bypassing the long and tedious RNA extraction step and reducing the time to viral detection. Methods: We evaluated three methods based on direct nasopharyngeal swab viral transmission medium (VTM) heating before the RT-qPCR: a) direct without additives; b) in a formamide-EDTA (FAE) buffer, c) in a RNAsnapTM buffer. Results: Although with a delay in cycle threshold compared to the gold-standard, we found consistent results in nasopharyngeal swab samples that were subject to a direct 70°C incubation for 10 min. Conclusions: Our findings provide valuable options to overcome any supply chain issue and help to increase the throughput of diagnostic tests, thereby complementing standard diagnosis..
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2020 |
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| Erschienen: |
2020 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:97 |
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| Enthalten in: |
International Journal of Infectious Diseases - 97(2020), Seite 66-68 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Julia Alcoba-Florez [VerfasserIn] |
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| Links: |
doi.org [kostenfrei] |
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| Themen: |
COVID-19 |
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| doi: |
10.1016/j.ijid.2020.05.099 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
DOAJ006911676 |
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| 520 | |a Objectives: The gold-standard COVID-19 diagnosis relies on detecting SARS-CoV-2 using RNA purification and one-step retrotranscription and quantitative PCR (RT-qPCR). Based on the urgent need for high-throughput screening, we tested the performance of three alternative, simple and affordable protocols to rapidly detect SARS-CoV-2, bypassing the long and tedious RNA extraction step and reducing the time to viral detection. Methods: We evaluated three methods based on direct nasopharyngeal swab viral transmission medium (VTM) heating before the RT-qPCR: a) direct without additives; b) in a formamide-EDTA (FAE) buffer, c) in a RNAsnapTM buffer. Results: Although with a delay in cycle threshold compared to the gold-standard, we found consistent results in nasopharyngeal swab samples that were subject to a direct 70°C incubation for 10 min. Conclusions: Our findings provide valuable options to overcome any supply chain issue and help to increase the throughput of diagnostic tests, thereby complementing standard diagnosis. | ||
| 650 | 4 | |a COVID-19 | |
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| 650 | 4 | |a RNA extraction | |
| 650 | 4 | |a fast protocols | |
| 653 | 0 | |a Infectious and parasitic diseases | |
| 700 | 0 | |a Rafaela González-Montelongo |e verfasserin |4 aut | |
| 700 | 0 | |a Antonio Íñigo-Campos |e verfasserin |4 aut | |
| 700 | 0 | |a Diego García-Martínez de Artola |e verfasserin |4 aut | |
| 700 | 0 | |a Helena Gil-Campesino |e verfasserin |4 aut | |
| 700 | 0 | |a The Microbiology Technical Support Team |e verfasserin |4 aut | |
| 700 | 0 | |a Laura Ciuffreda |e verfasserin |4 aut | |
| 700 | 0 | |a Agustín Valenzuela-Fernández |e verfasserin |4 aut | |
| 700 | 0 | |a Carlos Flores |e verfasserin |4 aut | |
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