Digital PCR based reference methods to improve quality of virus quantification

JRC scientists contributed to a research study assessing the robustness and transferability of digital PCR detection methods for the quantification of the human cytomegalovirus (CMV). The study demonstrated that carefully optimised digital PCR based detection methods can be transferred between laboratories delivering higher quality and more equivalent data compared to more classical quantitative PCR detection methods.

The use of quantitative PCR for quantification of pathogens is wide spread for routine applications. However, its dependence on the different reagents, the calibration materials and other experimental parameters is limiting the reproducibility of measurement results.
In collaboration within the Infect-Met consortium ('Metrology to Support Infectious Disease Diagnostics and Anti-microbial Resistance Monitoring' funded by the European Metrology Research Programme), JRC scientists have assessed the robustness and transferability of digital PCR detection methods for the quantification of the human cytomegalovirus (CMV). This research consortium investigated the potential of digital PCR for being used in reference methods with better performance characteristics in order to allow for a higher degree of standardisation of PCR based pathogen quantification methods. The digital PCR based CMV detection method was optimised and submitted to an inter-laboratory trial involving 4 laboratories of the consortium using different digital PCR platforms.
The study revealed that the repeatability and reproducibility of the optimised digital PCR method for CMV was superior to the characteristics of classical quantitative PCR methods. Therefore validated digital PCR based methods can be considered as candidate reference methods in a metrological framework.

Read more in:
J. Pavšič et al.: Inter-laboratory assessment of different digital PCR platforms for quantification of human cytomegalovirus DNA, Analytical and Bioanalytical Chemistry 409 (2017) 2601-2614,
doi: 10.1007/s00216-017-0206-0

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Inter-laboratory assessment of different digital PCR platforms for quantification of human cytomegalovirus DNA