Related Experiment Video
Updated: Dec 9, 2025

Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification
Published on: March 31, 2021
Use of a simplified sample processing step without RNA extraction for direct SARS-CoV-2 RT-PCR detection
Ruby Barza1, Parul Patel1, Linda Sabatini1
1NorthShore University HealthSystem, Department of Pathology and Laboratory Medicine, Evanston, IL, USA.
Abstract:
The severe acute respiratory syndrome coronavirus (SARS-CoV-2) pandemic has resulted in significant shortages of RT-PCR testing supplies including RNA extraction kits. The goal of our study was to determine if a simplified heat-RNA release method would provide comparable detection of SARS-CoV-2 without the need for nucleic acid extraction. RT-PCR results using the ChromaCode HDPCR™ SARS-CoV-2 were compared using the heat-RNA release method and an automated RNA extraction system (EMAG). The heat-RNA release method correctly identified 94 % (81/86 nasopharyngeal samples) that were positive for SARS-CoV-2. Five samples that were missed by heat-RNA release method had a mean Ct value: 35 using the automated extraction instrument, indicating a very low viral load. Our findings show that a simple heat-RNA release method is a reasonable alternative for the majority of COVID-19 positive patients and can help overcome the cost and availability issues of RNA extraction reagents.
Related Concept Videos
DNA Isolation
Real Time RT-PCR
The real-time quantification of the number of amplified products is...

