The CMV strains Towne, TR, and Toledo were serially diluted (IU/mL) into nuclease free water, prior to DNA extraction and PCR amplification. Resultant samples were assayed using a CRISPR-Cas12a based fluorescence detection assay. The limit of detection of the CMV gene UL123 using the CMV strain A) TR, B) Toledo, or C) Towne. The limit of detection of the CMV gene US28 using the CMV strain D) TR, E) Toledo, or F) Towne. N = 3 for all experiments. 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 < 0.0001), 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.