Applications of Digital PCR in Clinical Microbiology

Several potential advantages have emerged in recent years for the use of digital PCR (dPCR) instead of real-time quantitative PCR (qPCR) in clinical microbiology. As a direct, endpoint quantitation method, dPCR is not dependent on the use of calibration curves and is less affected by amplification inhibitors and suboptimal amplification efficiency. Some studies have shown improved accuracy and precision, with reduced variability based on target polymorphisms or differences in assay design. Viral quantitation is a particularly attractive application, as viral load assays have become integral to disease diagnosis, prediction of clinical outcome, monitoring of therapeutic response, and the initiation of preemptive treatment. The accurate quantitation of viral reservoirs and cellular copy number are of particular interest in the treatment of HIV. Beyond its use as a primary diagnostic method, digital PCR can be used to reduce variability among laboratories running qPCR through its use as a reference standard against which calibrators and other control materials can be measured.