Sensitive extraction-free SARS-CoV-2 RNA virus detection using a chelating resin

• Published in: iScience 2021-09, Vol.24 (9), p.102960-102960, Article 102960
• Main Authors: Guan, Bin, Frank, Karen M., Maldonado, José O., Beach, Margaret, Pelayo, Eileen, Warner, Blake M., Hufnagel, Robert B.
• Format: Article
• Language: English
• Subjects: Biological sciences; Biological sciences research methodologies; Molecular biology experimental approach; Virology
• ISSN: 2589-0042; 2589-0042
• DOI: 10.1016/j.isci.2021.102960

Current conventional detection of SARS-CoV-2 involves collection of a patient’s sample with a nasopharyngeal swab, storage of the swab during transport in a viral transport medium, extraction of RNA, and quantitative reverse transcription PCR (RT-qPCR). We developed a simplified preparation method using a chelating resin, Chelex, which obviates RNA extraction during viral testing. Direct detection RT-qPCR and digital droplet PCR were compared to the current conventional method with RNA extraction for simulated samples and patient specimens. The heat treatment in the presence of Chelex markedly improved detection sensitivity as compared to heat alone, and lack of RNA extraction shortens the overall diagnostic workflow. Furthermore, the initial sample heating step inactivates SARS-CoV-2 infectivity, thus improving workflow safety. This fast RNA preparation and detection method is versatile for a variety of samples, safe for testing personnel, and suitable for standard clinical collection and testing on high-throughput platforms. [Display omitted] •The COVID-19 pandemic caused supply shortages for diagnostic tests•Chelex resin preserves SARS-CoV-2 RNA in common buffers allowing direct RT-qPCR•The Chelex method presents a safe, economic, and sensitive test for RNA pathogens Biological sciences; Molecular biology experimental approach; Virology; Biological sciences research methodologies;