Absolute quantification of SARS-CoV-2 with Clarity Plus™ digital PCR
| Autor: | Huiyu Low, Shawn Yi Han Tan, Yan Ling Joy Pang, Sheng Yi Milton Kwek |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
2019-20 coronavirus outbreak
Wastewater-Based Epidemiological Monitoring Coronavirus disease 2019 (COVID-19) SARS-CoV-2 RT-dPCR assay Absolute quantification Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Wastewater Real-Time Polymerase Chain Reaction General Biochemistry Genetics and Molecular Biology Article Medicine Humans Digital polymerase chain reaction In patient Sample partitioning Molecular Biology business.industry SARS-CoV-2 COVID-19 Virology Orders of magnitude (mass) Clarity Plus™ dPCR Absolute viral quantification RNA Viral Target gene business |
| Zdroj: | Methods (San Diego, Calif.) |
| ISSN: | 1095-9130 1046-2023 |
| Popis: | In recent years, the usage of digital polymerase chain reaction (dPCR) for various clinical applications has increased exponentially. In this study, a dPCR assay optimized on the Clarity Plus™ dPCR system was evaluated for the absolute quantification of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of the global coronavirus disease 2019 (COVID-19) outbreak. The assay demonstrated good inter- and intra- assay precision, accuracy, as well as excellent linearity across a range of over 6 orders of magnitude for target gene quantification. In addition, a comparison of the assay on both dPCR and qPCR platforms revealed that dPCR exhibited a slightly higher sensitivity compared to its qPCR counterpart when quantifying SARS-CoV-2 at a lower concentration. Overall, the results showed that the dPCR assay is a reliable and effective approach for the absolute quantification of SARS-CoV-2 and can be a valuable molecular tool in clinical applications such as detecting low viral loads in patients as well as in wastewater surveillance of COVID-19. |
| Databáze: | OpenAIRE |
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