Abstract

Droplet Digital PCR: Applications in Diagnostic and Research Virology

Keith Jerome, Professor, University of Washington

Digital PCR offers several theoretical advantages over qPCR that suggest it might prove useful in clinical virology laboratories. We have applied the Bio-Rad droplet digital PCR system to several diagnostic virology applications. These studies confirm the extreme precision of dPCR quantitation, and reveal that dPCR is more resistant to inhibition than traditional qPCR. The precision of dPCR is especially useful in ratiometric assays, such as for the diagnosis of chromosomally integrated HHV-6. Conversely, for applications such as cytomegalovirus quantitation, the improved precision over qPCR does not clearly translate to clinical benefit. Precise quantitation of virus may be more useful at this point in research studies, such as evaluation of the HIV reservoir during eradication efforts. Finally, the current early generations of dPCR instrumentation have lower total throughput than the more mature qPCR platforms, and therefore traditional qPCR continues to play a role in diagnostic virology.