RT-RPA-Cas12a-based assay facilitates the discrimination of SARS-CoV-2 variants of concern

• DOI: 10.1016/j.snb.2023.133433
• Publication Date: 2023-01-26T23:09:35Z
• Authors: Guiyue Tang; Zilong Zhang; Wei Tan; Fei Long; Jingxian Sun; Yingying Li; Siwei Zou; Yujiao Yang; Kezhu Cai; Shenwei Li; Zhiyi Wang; Jiakun Liu; Guobing Mao; Yingxin Ma; Guo-Ping Zhao; Zhen-Gan Tian; Wei Zhao
• Keywords: 2019-20 coronavirus outbreak

Timely and accurate detection of SARS-CoV-2 variants concern (VOCs) is urgently needed for pandemic surveillance control. Great efforts have been made from a mass scientists in increasing the sensitivity operability, reducing turn-around time cost. Here, we report nucleic acid testing-based method aiming to detect discriminate mutations by combining RT-RPA CRISPR-Cas12a detecting assays (RRCd). With limit 10 copies RNA/reaction, RRCd was validated 194 clinical samples, showing 89% positive predictive agreement 100% negative agreement, respectively. Critically, using specific crRNAs, representatives single nucleotide polymorphisms small deletions VOCs including N501Y, T478K ΔH69-V70 were discriminated RRCd, demonstrating specificity samples with Ct < 33. The completes within 65 min could offer visible results without any electrical devices, which probably facilitate point-of-care testing other epidemic viruses.