Abstract
The increasing risk from viral outbreaks such as the ongoing COVID-19 pandemic exacerbates the need for rapid, affordable and sensitive methods for virus detection, identification and quantification; however, existing methods for detecting virus particles in biological samples usually depend on multistep protocols that take considerable time to yield a result. Here, we introduce a rapid fluorescence in situ hybridization (FISH) protocol capable of detecting influenza virus, avian infectious bronchitis virus and SARS-CoV-2 specifically and quantitatively in approximately 20 min, in virus cultures, combined nasal and throat swabs with added virus and likely patient samples without previous purification. This fast and facile workflow can be adapted both as a lab technique and a future diagnostic tool in enveloped viruses with an accessible genome.
Highlights
The increasing risk from viral outbreaks such as the ongoing COVID-19 pandemic exacerbates the need for rapid, affordable and sensitive methods for virus detection, identification and quantification; existing methods for detecting virus particles in biological samples usually depend on multistep protocols that take considerable time to yield a result
We introduce a rapid viral fluorescence in situ hybridization (FISH) protocol for the detection of virus particles; the development of this improved assay depended on a systematic analysis of the efficiency of the hybridization reaction as we reduced the number of experimental steps and the hybridization time
We explored the possibility of using a rapid FISH protocol for the detection of virus particles in culture supernatants and clinical samples
Summary
The increasing risk from viral outbreaks such as the ongoing COVID-19 pandemic exacerbates the need for rapid, affordable and sensitive methods for virus detection, identification and quantification; existing methods for detecting virus particles in biological samples usually depend on multistep protocols that take considerable time to yield a result. We introduce a rapid fluorescence in situ hybridization (FISH) protocol capable of detecting influenza virus, avian infectious bronchitis virus and SARS-CoV-2 and quantitatively in approximately 20 min, in virus cultures, combined nasal and throat swabs with added virus and likely patient samples without previous purification. This fast and facile workflow can be adapted both as a lab technique and a future diagnostic tool in enveloped viruses with an accessible genome. By increasing the number of fluorophores that bind one RNA transcript, detection of single transcripts became possible, giving rise to Scientific Reports | (2021) 11:19579
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