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Related Experiment Video

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Analytical Validation of a Pan-Cancer Next-Generation Sequencing Assay for In-House Liquid Biopsy Testing: An

Gaëlle Lescuyer1, Alexandre Harlé2, Hari Shankar Kumar3

  • 1Laboratoire de Biologie Médicale Multi-Sites, Department of Biochemistry and Molecular Biology, University of Lyon 1, Institute of Pharmaceutical and Biological Sciences, Center for Innovation in Cancerology of Lyon EA 3738, Lyon-Sud Hospital, Hospices Civils de Lyon, Pierre-Bénite, France.

The Journal of Molecular Diagnostics : JMD
|September 27, 2025
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Summary

The Hedera Profiling 2 (HP2) assay demonstrates high sensitivity and specificity for detecting tumor genetic alterations in liquid biopsies. This next-generation sequencing test shows strong potential for precision oncology in decentralized labs.

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Area of Science:

  • Oncology
  • Molecular Pathology
  • Genetics

Background:

  • Liquid biopsy assays offer a less invasive method for cancer genetic profiling compared to tissue biopsies.
  • Detecting low-frequency tumor variants in circulating DNA requires highly sensitive and specific assays.

Purpose of the Study:

  • To analytically evaluate the performance of the Hedera Profiling 2 (HP2) circulating tumor DNA test panel.
  • To assess HP2's capability in detecting various somatic alterations in cell-free DNA.

Main Methods:

  • Hybrid capture-based next-generation sequencing assay covering 32 genes.
  • Evaluation using reference standards and 137 precharacterized clinical samples.
  • Detection of single-nucleotide variants (SNVs), insertions/deletions (Indels), fusions, copy number variations, and microsatellite instability.

Main Results:

  • Sensitivity and specificity for SNVs/Indels were 96.92% and 99.67% at 0.5% allele frequency.
  • 100% sensitivity was achieved for fusion detection.
  • High concordance (94% for ESMO I) with orthogonal methods for SNV/Indel detection in clinical samples.

Conclusions:

  • The HP2 assay exhibits robust analytical performance for liquid biopsy testing.
  • HP2 demonstrates potential as a sensitive, efficient, pan-cancer assay for decentralized molecular pathology labs.