Diagnostic efficiency of RPA/RAA integrated CRISPR-Cas technique for COVID-19: A systematic review and meta-analysis

Abstract

Objective To evaluate the diagnostic value of recombinase polymerase/ aided amplification (RPA/RAA) integrated clustered regularly interspaced short palindromic repeats (CRISPR) in the diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Methods We searched relevant literature on CRISPR technology for COVID-19 diagnosis using "novel coronavirus", "clustered regularly interspaced short palindromic repeats" and "RPA/RAA" as subject terms in PubMed, Cochrane, Web of Science, and Embase databases. Further, we performed a meta-analysis after screening the literature, quality assessment, and data extraction. Results The pooled sensitivity, specificity and a rea under the summary receiver operator characteristic curve (AUC) were 0.98 [95% confidence interval (CI):0.97–0.99], 0.99 (95% CI: 0.97–1.00) and 1.00 (95% CI: 0.98–1.00), respectively. For CRISPR-associated (Cas) proteins-12, the sensitivity, specificity was 0.98 (95% CI: 0.96–1.00), 1.00 (95% CI: 0.99–1.00), respectively. For Cas13, the sensitivity and specificity were 0.99 (95% CI: 0.97–1.00) and 0.95 (95% CI: 0.91–1.00). The positive likelihood ratio (PLR) was 183.2 (95% CI: 28.8, 1166.8); the negative likelihood ratio (NLR) was 0.02 (95% CI: 0.01, 0.03). Conclusion RPA/RAA integrated with CRISPR technology is used to diagnose coronavirus disease-19 (COVID-19) with high accuracy and can be used for large-scale population screening.

Advances in the application of recombinase-aided amplification combined with CRISPR-Cas technology in quick detection of pathogenic microbes

“…The sensitivity of the RAA-Cas13a method can reach 10 copies of RNA molecules/reaction (Li et al, 2022). It summarizes all important reports about the application of RAA combined with CRISPR-Cas12a/Cas13a in the quick detection of pathogenic microbes (Table 1) (Li et al, 2021;Wang et al, 2022c;Zhang et al, 2022b;Chen et al, 2022;Fang et al, 2022;Qian et al, 2022;Wei et al, 2022;Xing et al, 2023;Zhao et al, 2023).…”

Advancements in the synergy of isothermal amplification and CRISPR-cas technologies for pathogen detection

“…The future holds promise for further advancements, including potential integration with other detection methods or the development of cost-effective materials. Compared to combining CRISPR with LAMP, RPA, and RAA, the former offers greater convenience and minimizes the occurrence of false positives ( Zhang X. et al, 2022 ). It is anticipated that as RPA and RAA technologies are further explored and refined, they will assume even more prominent roles in the future of molecular diagnostics.…”

“…It notably enhances detection sensitivity, making it particularly well-suited for on-site viral nucleic acid detection. However, it is worth noting that the inclusion of an additional CRISPR step may extend the assay’s reaction time, increase costs, and introduce complexity ( Zhang X. et al, 2022 ).…”