What question did this study set out to answer?

The research aims to enhance the sensitivity and accuracy of liquid biopsy by improving cfDNA extraction and mutation detection methods.

April 5, 2026

Abstract 7837: Enhanced sensitivity and scalability in liquid biopsy through integrated high-volume cfDNA extraction and droplet digital PCR mutation detection.

Key Points

The research aims to enhance the sensitivity and accuracy of liquid biopsy by improving cfDNA extraction and mutation detection methods.
Used nRichDX Revolution cfDNA Max20 Kit for high-volume extraction of plasma cfDNA.
Compared with Qiagen QIAamp Circulating Nucleic Acid Kit for efficiency.
Measured cfDNA concentration with Qubit fluorometry.
Quantified mutation abundance using Bio-Rad QX600 Droplet Digital PCR System.
Evaluated linearity, recovery efficiency, and reproducibility across varying input volumes.
nRichDX Kit showed linear cfDNA recovery across 1-15 mL plasma inputs (R2 = 0.98).
Demonstrated 15-30% higher mutation detection sensitivity compared to Qiagen Kit.
Preserved short cfDNA fragments critical for mutation detection, improving signal-to-background ratios.
Replicate variability remained below 10%, indicating robust precision.

Abstract

Abstract Introduction: Detecting rare tumor-derived variants in plasma requires both efficient recovery of cell-free DNA (cfDNA) and highly quantitative mutation analysis. Conventional extraction methods are limited by low input capacity and potential bias toward longer fragments, which can compromise sensitivity for low-frequency alleles. Methods: Plasma cfDNA was extracted using the nRichDX Revolution cfDNA Max20 Kit, a high-volume magnetic-bead-based system, and then compared with the Qiagen QIAamp Circulating Nucleic Acid Kit. Pooled human plasma (1-15 mL, n = 3) and contrived samples spiked with 1% EGFR E746A750delELREA (COSM6223) and KRAS G12C (COSM516) mutant gBlocks were processed per manufacturer’s protocols. cfDNA concentration was measured by Qubit fluorometry, and mutation fractional abundance was quantified using the Bio-Rad QX600™ Droplet Digital™ PCR System (ddPCR™). Linearity, recovery efficiency, and reproducibility were evaluated across input volumes. Results: The nRichDX Revolution cfDNA Max20 Kit demonstrated linear cfDNA recovery across 1-15 mL plasma inputs (R2 = 0. 98) and maintains scalability up to 50 mL without reconcentration or parallel extractions. When coupled with the Bio-Rad ddPCR System, mutation analysis consistently detected 1% variant allele frequencies, yielding 15-30% higher measured fractional abundance compared to the Qiagen QIAamp Circulating Nucleic Acid Kit, which uses carrier RNA. The nRichDX workflow preserved short cfDNA fragments critical for mutation detection, improving signal-to-background ratios and enhancing analytical sensitivity. Replicate variability remained below 10%, confirming robust precision across input volumes. Conclusions: Integration of the nRichDX Revolution cfDNA Max20 Kit with the Bio-Rad QX600 ddPCR System enhances analytical sensitivity, recovery accuracy, and scalability for cfDNA-based liquid biopsy applications. By maintaining cfDNA integrity and eliminating carrier RNA interference, this workflow enables the reliable quantification of low-frequency variants, supporting clinical research in early detection, therapy monitoring, and minimal residual disease assessment. Citation Format: Nafiseh Jafari, Andrew Partner, Necip Mehmet, Nish Kumar, Prithwish Pal, Jason Saenz, Carlos Hernandez, Daniel Cedeno, Cameron Van Dieren, Mayer Saidian. Enhanced sensitivity and scalability in liquid biopsy through integrated high-volume cfDNA extraction and droplet digital PCR mutation detection abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts) ; 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86 (7 Suppl): Abstract nr 7837.

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