Human Coronavirus 229E Remains Infectious on Common Touch Surface Materials

Warnes, Sarah L.; Little, Zoë R.; Keevil, C. William

rose gel using a GelRed nucleic acid prestaining kit (Biotium, United Kingdom) according to the manufacturer's instructions. The staining intensity is reduced because GelRed binds to single-stranded RNA (ssRNA) approximately half as much as to double-stranded nucleic acid. DNA ladders were supplied by Bioline. Gels were observed and photographed using GeneSnap software and a Syngene UV light box. Detection and quantification of a 139-bp region of the coronavirus nonstructural gene encoding nsp4 in virus exposed to copper and brass surfaces. To determine if exposure to copper affected the viral genome at the gene level, a 139-bp region of the gene encoding nsp4 (within polyprotein 1ab replicase) was investigated using a One-Step real-time quantitative PCR (RTqPCR) diagnostic kit (supplied by PrimerDesign, United Kingdom). The kit is based on sequences from HuCoV-229E (GenBank accession number NC_002645; anchor nucleotide position 8205). Amplification was performed on a BioRad iQ5 cycler, and standard curves were prepared from known copy number standards to determine copy numbers in test samples. PCR products were analyzed by gel electrophoresis as described above.