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.