Inhibition of polymerase chain reaction: Pathogen-specific controls are better than human gene amplification.

PCR inhibition is frequent in medical microbiology routine practice and may lead to false-negative results; however there is no consensus on how to detect it. Pathogen-specific and human gene amplifications are widely used to detect PCR inhibition. We aimed at comparing the value of PCR inhibitor detection using these two methods. We analysed Cp shifts (ΔCp) obtained from qPCRs targeting either the albumin gene or the pathogen-specific sequence used in two laboratory-developed microbiological qPCR assays. 3152 samples including various matrixes were included. Pathogen-specific amplification and albumin qPCR identified 62/3152 samples (2.0%), and 409/3152 (13.0%) samples, respectively, as inhibited. Only 16 samples were detected using both methods. In addition, the use of the Youden's index failed to determine adequate Cp thresholds for albumin qPCR, even when we distinguished among the different sample matrixes. qPCR targeting the albumin gene therefore appears not adequate to identify the presence of PCR inhibitors in microbiological PCR assays. Our data may be extrapolated to other heterologous targets and should discourage their use to assess the presence of PCR inhibition in microbiological PCR assays.

Déclaration de conflit d'intérêts

We acknowledge the financial support of the "RSI Assurance Maladie Professions Libérales - Provinces. C.A.M.P.L.P." for buying the LightCycler 480 (Roche®) real-time PCR equipment. Since this funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript; this does not alter our adherence to PLOS ONE policies on sharing data and materials.