Fig. 4: Sensitive and specific detection of HIV and HCV RNA using the CARRD assay.
From: CRISPR anti-tag-mediated room-temperature RNA detection using CRISPR/Cas13a
Comparison of a HIV detection and b HCV detection by the CARRD assay with the HP and conventional CRISPR/Cas13a assay without the HP. The ΔFluorescence intensity (FTarget RNA – FControl) of the CARRD assay (orange) and conventional CRISPR/Cas13a (green) at t = 60 min for various concentrations of the a HIV RNA and b HCV RNA targets (n = 3 and data represent mean ± s.d. of three technical replicates). The CARRD assay displayed superior sensitivity compared to the conventional Cas13a/crRNA reaction. c Time-dependent fluorescence signal changes of the CARRD assay (red) and conventional Cas13a/HIV crRNA (gray). The target HIV RNA concentration is 100 fM. Each curve was adjusted by subtracting the control signal (n = 3 and data represent mean ± s.d). d Specificity test using Cas13a/HIV crRNA and HIV 14D HP (black), and Cas13a/HCV crRNA and HCV 14D HP (orange) by the CARRD assay. Fluorescence intensity was measured at t = 60 min. For the Cas13a/HIV crRNA reaction; [HIV RNA] = 1 pM, [HCV RNA] = 10 pM, [HIV 14D HP] = 5 nM. For the Cas13a/HCV crRNA reaction; [HCV RNA] = 1 pM, [HIV RNA] = 10 pM, [HCV 14D HP] = 1 nM (n = 3 and data represent mean ± s.d. of three technical replicates). Source data are provided as a Source Data file.
