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Accreditation and Quality Assurance

, Volume 20, Issue 2, pp 75–83 | Cite as

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

  • Steffen UhligEmail author
  • Kirstin Frost
  • Bertrand Colson
  • Kirsten Simon
  • Dietrich Mäde
  • Ralf Reiting
  • Petra Gowik
  • Lutz Grohmann
General Paper

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.

Keywords

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

References

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Steffen Uhlig
    • 1
    Email author
  • Kirstin Frost
    • 1
  • Bertrand Colson
    • 1
  • Kirsten Simon
    • 1
  • Dietrich Mäde
    • 2
  • Ralf Reiting
    • 3
  • Petra Gowik
    • 4
  • Lutz Grohmann
    • 4
  1. 1.QuoData GmbHDresdenGermany
  2. 2.State Office for Consumer ProtectionHalle (Saale)Germany
  3. 3.Hessian State LaboratoryKasselGermany
  4. 4.Federal Office of Consumer Protection and Food SafetyBerlinGermany

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