5∣CONCLUSION
In summary, we have developed a CRISPR/Cas12a-based visual approach for SARS-CoV-2 detection. The visual detection process could be completed within 15 min. The LOD of ORF1ab gene and N gene of SARS-CoV-2 were 70 copies and 500 copies, respectively.
The VAPD assay was proved to be highly specific by assaying other respiratory virus standard substances and 60 clinical samples. No cross-reactivity was observed with those samples and the overall sensitivity and specificity of the VDAP compared to RT-PCR for detection SARS-CoV-2 were 93.7% and 100.0%, respectively. This simple, fast, high-specific, and convenient method can be a suitable tool to detect SARS-CoV-2 in hospital emergency, community, and primary care settings. However, this assay still requires improvement, particularly for the RNA extraction step. In the future, our group will conduct work on combining the VDAP assay with microfluidic technology to achieve point of care testing of the SARS-CoV-2.