CRISPR-Based Approaches for Efficient and Accurate Detection of SARS-CoV-2

Wancun Zhang, PhD; Kangbo Liu, MD; Pin Zhang, MD; Weyland Cheng, PhD; Linfei Li, MD; Fan Zhang, MD; Zhidan Yu, PhD; Lifeng Li, PhD; Xianwei Zhang, PhD

Disclosures

Lab Med. 2021;52(2):116-121. 

In This Article

Rapid Detection of SARS-CoV-2

The establishment of the SARS-CoV-2 rapid method is essential for responding to the outbreak of SARS-CoV-2. The IgG/IgM test kit has a short turnaround time with no specific requirements for additional equipment or skilled technicians, and it can be used as a point-of-care diagnosis method. However, the IgG/IgM test kit has a high rate of false positives and is not suitable for clinical use alone. It has been recommended that the IgG/IgM test kit could likely remedy false negatives inherent in respiratory swab specimens and could be administered as a complementary option to RT-PCR.[44,45] The CRISPR-based approach requires only 40 minutes for the entire detection process. However, the RT-PCR-based approach requires approximately 1.5 hours for a completion run of the PCR program. The mNGS method takes approximately 20 hours, which includes 8 hours of library preparation, 10 hours of sequencing, and 2 hours of bioinformatic analysis. Therefore, CRISPR-nCoV presents a significant advantage in effective turnaround time over RT-PCR and mNGS.[11] The Cas12-based lateral flow assay reported by Broughton et al[16] can be completed within 1 hour, as can the CRISPR-based SHERLOCK technique for the detection of SARS-CoV-2.[25] As previously mentioned, the STOP approach developed by Joung et al[27] returns results in 70 minutes using lateral flow readout and in 40 minutes using fluorescence readout. Therefore, a CRISPR-based approach can be used for the rapid detection of SARS-CoV-2.

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