CMV was serially diluted (IU/mL) into human urine, followed by DNA extraction and PCR amplification before CRISPR-based fluorescence detection. Limit of detection of UL123 targets using A) TR, B) Toledo, or C) Towne. Limit of detection of US28 targets using D) TR, E) Toledo, or F) Towne. N = 3. Positive is calculated by the mean of the NTC + (3*SD) and is indicated by the dotted line. *(P < 0.05), **(P < 0.01), ***(P < 0.001), ****(P < 0.001). Significance determined using a one-way ANOVA with a Dunnett’s multiple comparison test single pooled variance. NTC = no template control. a.u. = arbitrary units. IU = infectious units.