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™ 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.
【저자키워드】 COVID-19, Absolute viral quantification, Clarity Plus™ dPCR, SARS-CoV-2 RT-dPCR assay, Sample partitioning., 【초록키워드】 coronavirus disease, SARS-CoV-2, Coronavirus disease 2019, coronavirus, severe acute respiratory syndrome Coronavirus, polymerase chain reaction, sensitivity, Surveillance, Accuracy, Viral load, outbreak, qPCR, Patient, molecular, quantification, platform, CLARITY, Concentration, target gene, causative agent, Chain Reaction, acute respiratory syndrome, Precision, acute respiratory syndrome coronavirus, acute respiratory syndrome coronavirus 2, Usage, approach, effective, polymerase chain, Sample, addition, evaluated, exhibited, demonstrated, magnitude, 【제목키워드】 SARS-CoV-2, digital PCR, quantification,