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. 2024 Feb 13;13(3):837–850. doi: 10.1021/acssynbio.3c00655

Figure 3.

Figure 3

CRISPRD-based sensitive and specific detection of synthetic HPV16 and HPV18 DNA. (a) LoD for HPV16 with primer set C′ and gRNA #99 with AapCas12b. (b) Specificity of the CRISPRD reaction for HPV16 detection. The reaction mixture was incubated with 1 ng of HPV16, HPV18, gDNA, or RNase P (single target detection reactions). (c) HPV16 synthetic DNA fragments were spiked into 100 ng/μL gDNA to a final concentration of 1 ng/μL. (d) LoD for HPV18 using primer set F and gRNA #92. (e) Specificity of the CRISPRD reaction was determined for HPV18 detection. The reaction was incubated with 1 ng of HPV16, HPV18, gDNA, or RNase P (single target detection reactions). (f) HPV18 synthetic DNA was spiked into 100 ng/μL gDNA to a final concentration of 1 ng/μL. (g) LoD for RNase P with primer set Pop7 and gRNA #99 with TccCas13a. (h) Specificity of the CRISPRD reaction for RNase P detection. The reaction mixture was incubated with 1 ng of HPV16, HPV18, or RNase P (single target detection reactions). (i) Detection of RNase P in gDNA: 10 ng of gDNA was incubated in the CRISPRD reaction for RNase P detection. NTC: No-template control, RFU: relative fluorescence units, Cycle: 2 min (two-tailed Student t test; n.s., not significant; *P < 0.05 1;**P < 0.01; ***P < 0.001; ***P < 0.0001; All plots show mean ± SD for n = 3 replicates).