Validation of qualitative PCR methods within a single laboratory
This web service calculates the POD curve and the limit of detection LOD95%, i.e. the number of copies of the target DNA sequence required to ensure a 95 % probability of detection.
The model is closely related to the relative limit of detection approach as described in ISO Standard 16140-2:2014 as well as in Uhlig et al. (2014).
It is recommended to consider 6 dilution levels (e.g. 0.1, 1, 2, 5, 10 and 20 copies per 5 µL) and 12 PCR replicates per dilution level in order to obtain a statistically reliable POD curve from which the LOD95% is derived.