Mutation Detection Using Digital PCR | Thermo Fisher Scientific

dPCR enables quantitative detection of rare sequences in cancer and disease

Graphic image depicting cell-free DNA circulating in the blood stream

Digital PCR (dPCR) is a leading technology for detection and quantification of rare mutations and target sequences in cancer and disease. With the Applied Biosystems QuantStudio Absolute Q Digital PCR System, researchers can quickly and easily quantify relevant targets by leveraging a simple dPCR workflow backed by the trusted performance of TaqMan probe-based assays.

Benefits of digital PCR for rare mutation detection

dPCR can detect rare targets with mutation allele frequencies (MAFs) as low as 0.1%

Precise quantification of rare, single-nucleotide polymorphisms (SNPs) and other mutations over background requires highly sensitive analysis. Because dPCR effectively enriches low-level targets by partitioning samples into individual microchamber reactions, it is uniquely suited for applications requiring quantitative rare target detection, such as liquid biopsy analysis.

Technical note: Rare-target quantification on the the Absolute Q Digital PCR System

quantstudio-absolute-q-dPCR-rare-mutation

Figure 1. A demonstration of the ability of the QuantStudio Absolute Q dPCR System to detect mutant KRAS p.G12V alleles (purple dots) against a background of 99.9% wild type KRAS alleles (green dots) using the KRAS 520 assay and (orange dots) using the KRAS 520 assay. Green dots represent dPCR microchambers positive for both alleles.

dPCR increases sensitivity for rare target detection over conventional method

Absolute quantification using dPCR eliminates the need for a standard curve, improving both the accuracy and reproducibility of rare-target concentration measurement. dPCR also enables detection of ultra-rare target by leveraging pooling data to overcome limitations of sample input volume and concentration.

Poster: Precision genotyping for EGFR+ NSCLC using multiplexed dPCR on cell-free DNA
Technical Note: Improve sensitivity for rare targets by pooling digital PCR data

Graph depicting dPCR and qPCR EGFR mutation panel results

Figure 2. Correlation of EGFR Multiplex Panel results from the Absolute Q dPCR System to the duplex results gathered from qARMS-PCR. Each point represents a donor sample for which data was generated with both methods.

dPCR is an established technology for rare mutation analysis

With the ability to detect very low fractions of mutant targets in a background of abundant wild-type targets, dPCR is a technology of choice for cancer and genetic disease researchers.

Read QuantStudio Absolute Q dPCR publications ›

Digital PCR for liquid biopsy analysis

Liquid biopsy testing enables non-invasive biomarker testing to identify and track oncogenic mutations. Liquid biopsies detect cell-free DNA (cfDNA) released from apoptotic and necrotic cells. cfDNA that is released from cancer cells is referred to as circulating tumor DNA, or ctDNA.

dPCR enables precise quantification of ctDNA from liquid biopsies

Characterizing ctDNA helps researchers detect cancer early, measure therapeutic response, quantify residual tumor burden, and monitor emerging resistance to potential therapies. Because ctDNA fragments are typically short and very low in concentration, the sensitivity of dPCR makes it an ideal tool for ctDNA studies.

Webinar: dPCR in liquid biopsy for management and detection of cancer

Absolute Q dPCR assays for liquid biopsy

Absolute Q Liquid Biopsy dPCR assays enable reproducible, specific detection of known somatic mutations. Each predesigned assay has been validated using controls to detect down to 0.1% variant allele frequency in genes relevant for cancer research.

Absolute Q Liquid Biopsy dPCR assays are:

Simple—preformulated and verified assay in a single tube, enabling you to immediately start your research
Fast—minimal hands-on time; results in 90 minutes when used with the QuantStudio Absolute Q Digital PCR System
Highly Sensitive—detect down to 0.1% variant allele frequency
Robust—analyze your data with confidence using verified assays backed by a performance guarantee*

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Figure 3. Quantification of cancer mutations using Absolute Q Liquid Biopsy Digital PCR assays on as QuantStudio Absolute Q Digital PCR System.

Absolute Q liquid biopsy assays ordering information

To order Absolute Q Liquid Biopsy dPCR assays, indicate the desired quantity in the appropriate assay row, then scroll to the bottom of the table and click 'Add to cart'. Follow in-cart instructions to complete your purchase.