Basic Concepts and Validation of Digital PCR Measurements.

Use of digital polymerase chain reaction (dPCR) technology is rapidly growing and diversifying into a range of areas in life science. The release of dPCR commercial systems has facilitated access, leading to recognition of the potential advantages compared to previous quantitative PCR technologies, and the scope for novel applications. The capability of dPCR to deliver unprecedented levels of precision, accuracy, and resolution in quantification of nucleic acids has triggered a strong interest by academia and the life sciences industry in use of this technology as a molecular diagnostic tool. However, the performance of dPCR, as for a "classical" PCR assay, essentially still relies on enzyme-based amplification of nucleic acid using specific reagents and instrumentation. This chapter describes basic concepts, key properties, and important factors to consider for the verification and validation of dPCR measurements.