Validation of qualitative PCR methods on the basis of mathematical–statistical modelling of the probability of detection

Abstract

A new model for the probability of detection (POD curve) for qualitative PCR methods examined in a method validation collaborative study is presented. The model allows the calculation of the POD curve and the limit of detection (LOD 95%), i.e. the number of copies of the target DNA sequence required to ensure 95 % probability of detection. The between-laboratory variability of the limit of detection is used to derive the between-laboratory reproducibility of the PCR method. The model is closely related to the approach for quantitative methods described in ISO 5725.2:2002, and the relative limit of detection approach described in the new standard ISO 16140-2:2014.

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Correspondence to Steffen Uhlig.

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Uhlig, S., Frost, K., Colson, B. et al. Validation of qualitative PCR methods on the basis of mathematical–statistical modelling of the probability of detection. Accred Qual Assur 20, 75–83 (2015). https://doi.org/10.1007/s00769-015-1112-9

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Keywords

  • Probability of detection (POD)
  • Collaborative study
  • Real-time PCR
  • Validation
  • Generalized linear mixed model (GLMM)
  • Complementary log–log model