A Comparative Study of Real-Time RT-PCR–Based SARS-CoV-2 Detection Methods and Its Application to Human-Derived and Surface Swabbed Material
| Autor: | Pål Johansen, Reto Lienhard, Stephan Nobbe, Agathe Duda, Elisa Bellini, Philipp P. Bosshard, Corinne Isabelle Stoffel, Aizhan Tastanova, Mitchell P. Levesque, Phil F. Cheng, Andreas Dzung |
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| Přispěvatelé: | University of Zurich, Levesque, Mitchell Paul |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Surface Properties viruses Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) 610 Medicine & health Biology medicine.disease_cause Real-Time Polymerase Chain Reaction Sensitivity and Specificity Article Pathology and Forensic Medicine 03 medical and health sciences 0302 clinical medicine medicine Humans False Positive Reactions False Negative Reactions Pandemics Digital droplet pcr Coronavirus Detection limit Reverse Transcriptase Polymerase Chain Reaction SARS-CoV-2 COVID-19 10177 Dermatology Clinic Gold standard (test) Virology Highly sensitive 2734 Pathology and Forensic Medicine 030104 developmental biology Emergency response Real-time polymerase chain reaction 030220 oncology & carcinogenesis COVID-19 Nucleic Acid Testing 1313 Molecular Medicine Feasibility Studies RNA Viral Molecular Medicine Smartphone Switzerland |
| Zdroj: | The Journal of Molecular Diagnostics : JMD |
| DOI: | 10.5167/uzh-203777 |
| Popis: | Real-time reverse transcription polymerase chain reaction (RT-PCR) remains a gold standard in detection of various viral diseases. In the COVID-19 pandemic, multiple RT-PCR based tests were developed to screen for viral infection. As an emergency response to growing testing demand, we established a SARS-CoV-2 PCR diagnostics platform for which we compared different commercial and in-house RT-PCR protocols. Four commercial (CDC 2019-nCoV, Applied BiosystemsTM 2019-nCoV Assay Kit v1 TF-SinglePlex, 2019-nCoV Assay Kit v2 TF-MultiPlex, and EURORealTime SARS-CoV-2), one customized (Institute Pasteur), and one in-house RT-PCR protocols were evaluated with 92 SARS-CoV-2 positive and 92 SARS-CoV-2 negative samples. Furthermore, economical and practical characteristics of these protocols were compared. Additionally, a highly sensitive digital droplet PCR (ddPCR) method was developed and application of RT- and ddPCR methods on SARS-CoV-2 environmental samples was examined. Very low limits of detection (1 or 2 viral copies/μL), high sensitivities (93.6-97.8%) and specificities (98.7-100%) for the tested RT-PCR protocols were found. Furthermore, the feasibility of downscaling two of the commercial protocols, which could optimize testing capacity was demonstrated. Tested commercial and customized RT-PCR detection kits show very good and comparable sensitivity, and specificity, and the kits could be further optimized for use on SARS-CoV-2 viral samples derived from human and surface swabbed samples. |
| Databáze: | OpenAIRE |
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