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

Lescuyer, Gaëlle; Harlé, Alexandre; Kumar, Hari Shankar; Constantoulakis, Pantelis; Pfarr, Nicole; Heitzer, Ellen; Michon, Clémence; Russo, Gianluca; Speel, Ernst-Jan M; Piecyk, Marie; Husson, Marie; Christopoulou, Georgia; Mayr, Eva-Maria; Koppermann, Mai-Lan; Passot, Christophe; Graf, Ricarda; Aoul, Anes Hadjadj; Bourdon, Violaine; Dubbink, Hendrikus J; van Marion, Ronald; Demers, Imke; Dingemans, Anne-Marie C; Troncone, Giancarlo; Pepe, Francesco; Muinelo-Romay, Laura; Díaz-Lagares, Ángel; Rodriguez-Casanova, Aitor; Lago Lestón, Ramón Manuel; Pathak, Deepak; Shah, Parth; Parillaud, Romain V; Martínez de Ilarduya, Oskar; Behr, Jonas; Rapin, Alexis; Vetterli, Thomas; Boppudi, Sanga Mitra; Malapelle, Umberto; Payen, Léa     «

Lescuyer, Gaëlle; Harlé, Alexandre; Kumar, Hari Shankar; Constantoulakis, Pantelis; Pfarr, Nicole; Heitzer, Ellen; Michon, Clémence; Russo, Gianluca; Speel, Ernst-Jan M; Piecyk, Marie; Husson, Marie; Christopoulou, Georgia; Mayr, Eva-Maria; Koppermann, Mai-Lan; Passot, Christophe; Graf, Ricarda; Aoul, Anes Hadjadj; Bourdon, Violaine; Dubbink, Hendrikus J; van Marion, Ronald; Demers, Imke; Dingemans, Anne-Marie C; Troncone, Giancarlo; Pepe, Francesco; Muinelo-Romay, Laura; Díaz-Lagares, Ángel; Ro...     »

Liquid biopsy assays are transforming precision oncology by providing a less invasive alternative to tissue biopsies. These assays screen for tumor genetic alterations in circulating free DNA, which typically requires detecting variants at low allele frequencies and, therefore, a high sensitivity and specificity. This international, multicenter analytical performance study evaluated the Hedera Profiling 2 circulating tumor DNA test panel (HP2), a hybrid capture-based next-generation sequencing assay for the detection of somatic alterations in circulating free DNA. Covering 32 genes, HP2 enables the detection of single-nucleotide variants (SNVs), insertions and deletions (Indels), fusions, copy number variations, and microsatellite instability status from a single DNA-only workflow. The analytical performance was assessed using reference standards and a diverse cohort of 137 clinical samples precharacterized by orthogonal methods. In reference standards with variants spiked in at 0.5% allele frequency, sensitivity and specificity were 96.92% and 99.67%, respectively, for SNVs/Indels and 100% for fusions. In clinical samples, SNV/Indel detection showed high concordance (94% for European Society for Medical Oncology Scale of Clinical Actionability for Molecular Targets level I variants) with orthogonal methods. Evidence for solid sensitivity was also found for copy number variation detection and microsatellite instability status determination. Overall, the HP2 assay showed significant potential as a sensitive and efficient pan-cancer test for liquid biopsy testing in a decentralized molecular pathology laboratory setting.      «

Liquid biopsy assays are transforming precision oncology by providing a less invasive alternative to tissue biopsies. These assays screen for tumor genetic alterations in circulating free DNA, which typically requires detecting variants at low allele frequencies and, therefore, a high sensitivity and specificity. This international, multicenter analytical performance study evaluated the Hedera Profiling 2 circulating tumor DNA test panel (HP2), a hybrid capture-based next-generation sequencing a...     »