The power of digital PCR in clinical studies

Poster

This poster was presented at the NextGen Omics UK virtual meeting, November 2020

Since becoming commercially available in 2011, digital PCR (dPCR) has been gaining in popularity as it enables nucleic acid quantification with superior accuracy and precision compared to quantitative PCR (qPCR). In brief, the technology consists of the following steps:

  1. The PCR reaction mixture is partitioned into thousands of water-in-oil droplets with target and background DNA randomly distributed among the reactions
  2. The target DNA is amplified by PCR using standard thermal cycling with fluorescent dyes or probes
  3. Each reaction provides a fluorescent positive or negative signal indicating that target DNA was present or absent in partitioning
  4. The fraction of positive droplets is used to calculate the target DNA concentration using Poisson correction

With data acquisition taking place at the end of the reaction and since absolute concentrations can be determined, data can be easily interpreted and compared across different samples and experiments, especially in a clinical setting. Additionally, dPCR doesn’t suffer from the inherent complications of qPCR that result from the need of a standard curve, and excels in the quantification of low abundant genes or small differences.

In this poster, we perform a quick comparison on qPCR versus dPCR, and outline the intrinsic potential of ddPCR in clinical research for 3 different applications:

  • Rare variant analysis in plasma samples
  • Small changes in alternative splicing
  • CAR-T studies

Download the Poster

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