Absolute quantification of SARS-CoV-2 with Clarity Plus™ digital PCR

Area of Science:

• Molecular Biology
• Clinical Diagnostics
• Virology

Background:

• Digital PCR (dPCR) is increasingly used in clinical settings.
• Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes COVID-19.
• Accurate quantification of SARS-CoV-2 is crucial for patient management and public health surveillance.

Purpose of the Study:

• To evaluate an optimized dPCR assay for absolute quantification of SARS-CoV-2.
• To assess the precision, accuracy, and linearity of the dPCR assay.
• To compare the sensitivity of dPCR versus qPCR for SARS-CoV-2 detection.

Main Methods:

• Optimization of a dPCR assay on the Clarity Plus™ dPCR system.
• Absolute quantification of SARS-CoV-2 using the dPCR assay.
• Inter- and intra-assay precision and accuracy assessment.
• Linearity testing across six orders of magnitude.
• Comparative analysis with quantitative PCR (qPCR) platforms.

Main Results:

• The dPCR assay demonstrated excellent inter- and intra-assay precision and accuracy.
• High linearity was observed for target gene quantification over a wide dynamic range (>6 orders of magnitude).
• dPCR showed slightly higher sensitivity than qPCR for low-concentration SARS-CoV-2 samples.
• The dPCR assay proved reliable for absolute quantification of SARS-CoV-2.

Conclusions:

• The dPCR assay is a robust and effective tool for absolute SARS-CoV-2 quantification.
• This method is valuable for detecting low viral loads in patients.
• The dPCR assay is suitable for wastewater surveillance of COVID-19.
• dPCR offers advantages in sensitivity for low viral concentration detection compared to qPCR.