Evaluation of extraction-free RT-qPCR methods for SARS-CoV-2 diagnostics

Alexander Domnich, Vanessa De Pace, Beatrice M. Pennati, Patrizia Caligiuri, Serena Varesano, Bianca Bruzzone, Andrea Orsi

Research output: Contribution to journalArticlepeer-review


Extraction-based real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR) is currently the “gold standard” in SARS-CoV-2 diagnostics. However, some extraction-free RT-qPCR techniques have recently been developed. In this study, we compared the sensitivity of traditional extraction-based, heated extraction-free, and unheated extraction-free RT-qPCR methods for SARS-CoV-2 detection in nasopharyngeal swabs from symptomatic individuals. The unheated extraction-free method showed perfect agreement with the standard extraction-based RT-qPCR. By contrast, the heat-treated technique was associated with an 8.2% false negativity rate. Unheated extraction-free RT-qPCR for the molecular diagnosis of SARS-CoV-2 is a valuable alternative to the traditional extraction-based methods and may accelerate turnaround times by about two hours.

Original languageEnglish
JournalArchives of Virology
Publication statusAccepted/In press - 2021

ASJC Scopus subject areas

  • Virology


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