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. 2018 May 16;13(5):e0197074. doi: 10.1371/journal.pone.0197074

Fig 1. Analytical sensitivity of the LN34 assay.

Fig 1

A. Result of limit of detection analysis using serial dilutions of artificial positive control RNA. Estimated RNA copy number was plotted against the percent of runs in which amplification was observed (blue dots). A sigmoidal curve was fit to the data to estimate the limit of detection as the RNA copy number corresponding to 95% assay success (black line, gray shading indicates 95% confidence interval). B. LN34 assay Ct value upon 10 fold serial dilutions of RNA extracted from six lyssavirus isolates. Average Ct value and 95% confidence intervals are shown; several points are artificially offset along the x axis to avoid overlap. Linear regression analysis revealed the following slopes: DUVV -3.58, LBV -3.37, RABV1–3.64, RABV2–3.84, RABV3–3.66, RABV4–3.56. LN34 assay diagnostic cut-off for positive samples (Ct 35) is highlighted in yellow. Assay failure (Ct 45) is shown in red.