Rapid Coronavirus Detection with Portable CRISPR-based Mobile Tests

The recent coronavirus disease caused by SARS-COV-2 has risen due to the challenges of identifying the asymptomatic patients and presymptomatic carriers of the virus which resulted in delayed isolation and massive global spread. The standard procedure involves a quantitative reverse transcription-polymerase chain reaction (RT-qPCR) which takes a longer time to process. This highlights the need for rapid testing that can detect the SARS-COV-2 genetic material.

In that regard, viral diagnostics may be of benefit from a successful bacterial strategy to destroy incoming bacteriophages and build an immunological memory, which is known as CRISPR. To achieve the high sensitivity needed for testing purposes, current CRISPR diagnostic strategies primarily rely on pre-amplification of target RNA for detection by a Cas protein. In a new study, a research group reported the proof of concept of a rapid CRISPR-Cas13a-based diagnostic assay for the direct detection of SARS-CoV-2 RNA.

For this, the researchers first needed to optimize Cas13 activation through careful selection of CRISPR RNA complexes, as well as to develop a sensitive and transportable fluorescence detection system for this novel assay. The simplicity of the approach was demonstrated by measuring fluorescence with a mobile phone camera in a close-packed device, consisting of low-cost laser lightning and collection optics. A key development of this approach is a successful demonstration of how these combinations of crRNAs can increase the sensitivity of Cas13a direct detection by including more Cas13a per target RNA.

Another key to note is the possibility to translate the fluorescent signal into viral loads directly. Unlike previous diagnostic assays, this one does not require pre-amplification of the viral genome. Mobile phone cameras became highly sensitive tools, making them attractive devices for point-of-care disease diagnosis, especially in low-resource regions. Further, direct detection by Cas13a may be quickly modified to target the next emerging respiratory pathogen – hopefully in time to help contain the global spread and another pandemic.

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Ref link: https://www.medrxiv.org/content/10.1101/2020.09.28.20201947v1