Digital PCR in Clinical Microbiology: Advantages, Limitations, and Practical Considerations
Digital PCR (dPCR) enables absolute nucleic acid quantification without standard curves, offering advantages over qPCR for applications requiring high precision at low copy numbers. This Clinical Chemistry review compares the two technologies across clinical microbiology use cases including viral load monitoring, resistance detection, and transplant-associated infection surveillance. While dPCR offers superior precision and tolerance to inhibitors, the authors note throughput limitations and higher per-reaction costs that laboratories must weigh for each application.
The original study
Digital PCR-An Emerging Technology with Broad Applications in Microbiology.
- Authors
- Salipante SJ, Jerome KR
- Journal
- Clinical chemistry
- Type
- Journal Article, Review
- PMID
- 31704712
Original abstract
BACKGROUND: The PCR and its variant, quantitative PCR (qPCR), have revolutionized the practice of clinical microbiology. Continued advancements in PCR have led to a new derivative, digital PCR (dPCR), which promises to address certain limitations inherent to qPCR. CONTENT: Here we highlight the important technical differences between qPCR and dPCR, and the potential advantages and disadvantages of each. We then review specific situations in which dPCR has been implemented in clinical microbiology and the results of such applications. Finally, we attempt to place dPCR in the context of other emerging technologies relevant to the clinical laboratory, including next-generation sequencing. SUMMARY: dPCR offers certain clear advantages over traditional qPCR, but these are to some degree offset by limitations of the technology, at least as currently practiced. Laboratories considering implementation of dPCR should carefully weigh the potential advantages and disadvantages of this powerful technique for each specific application planned.