This study presents a structured approach to aligning quantitative PCR (qPCR) and digital PCR (dPCR) data outputs to enable more consistent, cross-platform analysis. By applying standardized reporting units (copies/µL of eluate), implementing negative control–anchored thresholding, and leveraging multiplexing capabilities, the team evaluated sensitivity, precision, and overall performance between the two technologies.
Using identical sample extractions, the Absolute Q dPCR system demonstrated robust multiplex quantification with clear target separation and strong precision, while maintaining comparable detection rates to qPCR across a range of concentrations. Although qPCR showed a slightly lower limit of detection under the tested conditions, this difference was primarily attributed to input volume rather than platform limitations. With optimized input strategies, dPCR has the potential to match or exceed qPCR sensitivity while offering increased resilience to PCR inhibitors.
Overall, the findings highlight practical strategies—such as standardized unit reporting, controlled thresholding, and precise input accounting—that support reliable method transfer and platform selection. These approaches are particularly valuable for low-titer samples and complex multiplex assays, where consistency and reproducibility across analytical platforms are critical.
