Open AccessIncreased sensitivity using real-time dPCR for detection of SARS-CoV-2
Author(s) -
Kyra Duong,
Jiajia Ou,
Zhao-Liang Li,
Zhaoqing Lv,
Hao Dong,
Tao Hu,
Yunyun Zhang,
Ava Hanna,
Skyler Gordon,
Gogce Crynen,
Steven R. Head,
Phillip Ordoukhanian,
Yan Wang
Publication year - 2021
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/btn-2020-0133
Subject(s) - digital polymerase chain reaction , sensitivity (control systems) , context (archaeology) , taqman , computer science , chemistry , real time polymerase chain reaction , biology , polymerase chain reaction , paleontology , biochemistry , electronic engineering , engineering , gene
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.