Fig. 7: TRACE duplex detection of MPXV and RNase P gene.
From: Thermally programmed one-pot CRISPR assay for on-site pandemic surveillance
a Schematic of the TRACE duplex assay, indicating active duplex RPA reactions and CRISPR/Cas12a–mediated RNase P signal production in the 37 °C first stage, and subsequent RPA and Cas12a inactivation and formation of active Cas12b/gRNA complexes and MPXJ J2L signal production upon release of the ssRNA blocker in the 64 °C second stage. ΔF1 and ΔF2 indicate host RNase P- and MPXV J2L-specific signals produced in the first and second stages, respectively. Created in BioRender. Dong, Y. (https://BioRender.com/7yzlkxh). b ΔF2 signal (%) contributed by Cas12a/gRNA and Cas12b/gRNA complexes at the indicated temperatures (n = 3 samples/condition; detailed results shown in Supplementary Fig. 22). c Schematic of ΔF1 and ΔF2 signatures characteristic of samples containing detectable levels of either, both, or neither of the RNase P (293 cell extracts) or MPXV J2L targets (100 copies/test). The red triangle denotes the point of temperature transition. d Normalized ΔF1 signal detected for the indicated RNase P concentrations (n = 3/group). e Normalized ΔF1 and ΔF2 signal detected and target positivity rates in 293 cell nucleic acid extracts (103 ng/μL) spiked with the indicated MPXV J2L DNA concentrations. Created in BioRender. Dong, Y. (https://BioRender.com/rwlikje). f Normalized TRACE duplex assay RNase P ΔF1 and MPXV J2L ΔF2 signals detected in 12 MPXV-positive and 11 MPXV-negative clinical samples. Graphs indicate mean ± SD values. Source data are provided as a Source Data file.
