tetano
Editor, Senior Moderator
Methods
. 2021 Jul 14;S1046-2023(21)00184-5.
doi: 10.1016/j.ymeth.2021.07.005. Online ahead of print.
Absolute quantification of SARS-CoV-2 with Clarity Plus™ digital PCR
Shawn Yi Han Tan[SUP] 1 [/SUP], Sheng Yi Milton Kwek[SUP] 2 [/SUP], Huiyu Low[SUP] 2 [/SUP], Yan Ling Joy Pang[SUP] 3 [/SUP]
Affiliations
Abstract
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[SUP]TM[/SUP] 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.
Keywords: Absolute viral quantification; COVID-19; Clarity Plus™ dPCR; SARS-CoV-2 RT-dPCR assay; Sample partitioning.
. 2021 Jul 14;S1046-2023(21)00184-5.
doi: 10.1016/j.ymeth.2021.07.005. Online ahead of print.
Absolute quantification of SARS-CoV-2 with Clarity Plus™ digital PCR
Shawn Yi Han Tan[SUP] 1 [/SUP], Sheng Yi Milton Kwek[SUP] 2 [/SUP], Huiyu Low[SUP] 2 [/SUP], Yan Ling Joy Pang[SUP] 3 [/SUP]
Affiliations
- PMID: 34273478
- DOI: 10.1016/j.ymeth.2021.07.005
Abstract
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[SUP]TM[/SUP] 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.
Keywords: Absolute viral quantification; COVID-19; Clarity Plus™ dPCR; SARS-CoV-2 RT-dPCR assay; Sample partitioning.