Increased sensitivity using real-time dPCR for detection of SARS-CoV-2

Duong, Kyra; Ou, Jiajia; Li, Zhaoliang; Lv, Zhaoqing; Dong, Hao; Hu, Tao; Zhang, Yunyun; Hanna, Ava; Gordon, Skyler; Crynen, Gogce; Head, Steven R; Ordoukhanian, Phillip; Wang, Yan

A real-time dPCR system was developed to improve the sensitivity, specificity and quantification accuracy of end point dPCR. We compared three technologies – real-time qPCR, end point dPCR and real-time dPCR – in the context of SARS-CoV-2. Some improvement in limit of detection was obtained with end point dPCR compared with real-time qPCR, and the limit of detection was further improved with the newly developed real-time dPCR technology through removal of false-positive signals. Real-time dPCR showed increased linear dynamic range compared with end point dPCR based on quantitation from amplification curves. Real-time dPCR can improve the performance of TaqMan assays beyond real-time qPCR and end point dPCR with better sensitivity and specificity, absolute quantification and a wider linear range of detection.