Fig. 5: Evaluation of the analytical performance of SCOPE.

Real-time fluorescence curves were tested by (a) CPod and (b) plate reader respectively and (c) their end-point fluorescence intensity for detection of monkeypox virus were collected with the DNA target at the concentrations of 1000, 100, 10, 5, 1, 0.5, 0.2, and 0 copies/µL. Sample volume: 5 µL. *** means P < 0.001. ‘ns’ means no significance, P > 0.05. d Reproducibility of our CRISPR-based system for the detection of monkeypox virus DNA at the concentrations of 1, 0.5, 0.4, 0.3, 0.2, and 0 copies/µL with independently repeating for 20 times. Sample volume: 5 µL. e Table of positive rates of testing with different concentrations of monkeypox virus DNA in (d). f Validation of the cross-reactivity between monkeypox virus (MPXV) detection and other orthopox virus and other viruses with similar symptoms detection including cowpox, vaccinia, variola, herpes simplex virus (HSV), and varicella using SCOPE. Templates concentration: 10⁶ copies/µL, sample volume: 5 µL. **** means P < 0.0001. ‘n.d.’ means not detected. For (a–c) and (f), mean ± s.d. for 3 technical replicates. For (d), mean ± s.d. for 20 technical replicates. For (c, f), P values were calculated by two-tailed Student’s t-test.