Measurement uncertainty in calibration and compliancy testing of PCR and qPCR thermal cyclers
1 CYCLERtest BV, Rötscherweg 61, 6374 XW Landgraaf, the Netherlands
2 CelsiusLabs, Rötscherweg 61, 6374 XW Landgraaf, the Netherlands
* Corresponding author: firstname.lastname@example.org
Both testing and medical laboratories use thermal cycler and real-time thermal cycler equipment for Polymerase Chain Reaction (PCR) based DNA testing. These thermal cyclers require regular calibration to ensure proper functioning. Currently, a variety of thermal cycler calibration and verification methods exits. Most of these methods mimic the PCR reaction, are not traceable to the International System of Units (SI) and connected to substantial measurement uncertainties, impeding compliancy reporting. Therefore these methods are inapt for application in metrology. This paper discusses a universal temperature calibration method for thermal cyclers that can be applied to any brand and model PCR and qPCR# thermal cycler, irrespective of the heating and cooling technology used. The calibration method and low associated measurement uncertainties do allow meaningful compliancy reporting to meet the requirements of ISO/IEC 17025, ISO 15189, ILAC P10 and ILAC G8. In addition, the paper offers effective practical solutions to determine PCR method based specifications, rather than thermal cycler manufacturer specifications, to link the PCR method to the thermal cycler, in order to qualify if the instrument is suitable for the intended use.
© The Authors, published by EDP Sciences, 2017
This is an Open Access article distributed under the terms of the Creative Commons Attribution License 4.0, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.