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Updated: Dec 8, 2025

Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification
Published on: March 31, 2021
qRT-PCR Platforms for Diagnosing and Reporting SARS-CoV-2 Infection in Human Samples
Richard M Giadone1, Aditya Mithal1, Gregory J Miller1
1Center for Regenerative Medicine of Boston University and Boston Medical Center, Boston, MA 02118, USA.
Abstract:
The protocols herein outline the use of qRT-PCR to detect the presence of SARS-CoV-2 genomic RNA in patient samples. In order to cope with potential fluctuations in supply chain and testing demands and to enable expedient adaptation of reagents and assays on hand, we include details for three parallel methodologies (one- and two-step singleplex and one-step multiplex assays). The diagnostic platforms described can be easily adapted by basic science research laboratories for SARS-CoV-2 diagnostic testing with relatively short turnaround time. For complete details on the use and execution of this protocol, please refer to Vanuytsel et al. (2020).

