Fig. 4.
Calculation of the sample-based detection limit from datasets. (a) Key characteristics of the analysed samples and datasets. Upper panel, dataset size; mid panel, total RNA concentration; lower panel, LODmNGS. Calculations were performed with 30 datasets originated from diseased animals (Table 1) by combination of equations 2, 3, and 4. (b) Comparison of the LODmNGS (c/µl) and viral genome copy numbers (c/µl) as determined by RT-qPCR quantification of the samples. Red lines display the samples where LODmNGS > RT-qPCR c/µl. (c) Probability-based calculation of LODmNGS for the five samples labelled red in (b). Horizontal lines represent the adjusted α to meet the criteria LODmNGS < copy number determined by RT-qPCR. Vertical lines represent the calculated LODmNGS (c/µl) with the adjusted α. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)