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. 2023 Apr 23;25(7):428–437. doi: 10.1016/j.jmoldx.2023.03.009

Figure 2.

Figure 2

Real-time SHERLOCK (RT-SLK). A: The real-time SLK Direct process. B–F: Time to result (TTR, in minutes) in each sample. Samples not detected are labeled ND (not determined). B: Titration of extracted SARS-CoV-2 genomic RNA, ranging from 100,000 to 0.1 cp/μL. Each concentration was tested in triplicate. C and D: TTR of RNaseP control signal (RP) versus SARS-CoV-2 N/Orf1ab target (NO) in ANS/saline samples spiked with indicated concentrations of SARS-CoV-2 viral particles. C: ANS/saline samples were purified using the PureLink viral DNA/RNA kit (Thermo Fisher Scientific, Waltham, MA) then added to real-time SLK reaction. D: ANS/saline samples were treated with Proteinase K (New England BioLabs, Ipswich, MA), RNAsecure (InvitroGen, Waltham, MA), and heat and directly added to real-time SLK Direct reaction. E and F: Positive and negative clinical samples were tested using real-time SLK and the CDC's RT-qPCR assay; results are reported as TTR. Dotted lines indicate cutoff values for either assay, and samples that were not detected are labeled ND and considered negative. E: Clinical samples—36 NP/saline, 10 NP/viral transport medium, and 10 NP/universal transport medium—were purified using the PureLink viral DNA/RNA kit and added to the real-time SLK reaction. F: A total of 36 NP/saline clinical samples were treated with Proteinase K, RNAsecure, and heat and directly added to a real-time SLK Direct reaction. gRNA, genomic RNA.