Multiplex Detection of Clinically Relevant Mutations in Liquid Biopsies of Cancer Patients Using a Hybridization-Based Platform
Giménez-Capitán, Ana (Grup Quirónsalud (Barcelona, Catalunya))
Bracht, Jillian (Universitat Autònoma de Barcelona)
García Mosquera, Juan José (Grup Quirónsalud (Barcelona, Catalunya))
Jordana-Ariza, Núria (Grup Quirónsalud (Barcelona, Catalunya))
García, Beatriz (Grup Quirónsalud (Barcelona, Catalunya))
Garzón Ibáñez, Mónica (Grup Quirónsalud (Barcelona, Catalunya))
Mayo-De-Las-Casas, Clara (Grup Quirónsalud (Barcelona, Catalunya))
Viteri, Santiago (Grup Quirónsalud (Barcelona, Catalunya))
Martínez-Bueno, Alejandro (Grup Quirónsalud (Barcelona, Catalunya))
Aguilar-Hernández, Andrés (Grup Quirónsalud (Barcelona, Catalunya))
Sullivan, Ivana (Institut d'Investigació Biomèdica Sant Pau)
Johnson, Eric (NanoString Technologies)
Huang, Chun-Ying (NanoString Technologies)
Gerlach, Jay L. (NanoString Technologies)
Warren, Sarah (NanoString Technologies)
Beechem, Joseph M. (NanoString Technologies)
Teixidó, Cristina (Institut d'Investigacions Biomèdiques August Pi i Sunyer)
Rosell, Rafael (Institut Germans Trias i Pujol. Hospital Universitari Germans Trias i Pujol)
Reguart, Noemi (Hospital Clínic i Provincial de Barcelona)
Molina-Vila, Miguel Ángel. (Grup Quirónsalud (Barcelona, Catalunya))

Date:	2021
Abstract:	BACKGROUND: With the advent of precision oncology, liquid biopsies are quickly gaining acceptance in the clinical setting. However, in some cases, the amount of DNA isolated is insufficient for Next-Generation Sequencing (NGS) analysis. The nCounter platform could be an alternative, but it has never been explored for detection of clinically relevant alterations in fluids. METHODS: Circulating-free DNA (cfDNA) was purified from blood, cerebrospinal fluid, and ascites of patients with cancer and analyzed with the nCounter 3 D Single Nucleotide Variant (SNV) Solid Tumor Panel, which allows for detection of 97 driver mutations in 24 genes. RESULTS: Validation experiments revealed that the nCounter SNV panel could detect mutations at allelic fractions of 0. 02-2% in samples with 5 pg mutant DNA/mL. In a retrospective analysis of 70 cfDNAs from patients with cancer, the panel successfully detected EGFR, KRAS, BRAF, PIK3CA, and NRAS mutations when compared with previous genotyping in the same liquid biopsies and paired tumor tissues [Cohen kappa of 0. 96 (CI = 0. 92-1. 00) and 0. 90 (CI = 0. 74-1. 00), respectively]. In a prospective study including 91 liquid biopsies from patients with different malignancies, 90 yielded valid results with the SNV panel and mutations in EGFR, KRAS, BRAF, PIK3CA, TP53, NFE2L2, CTNNB1, ALK, FBXW7, and PTEN were found. Finally, serial liquid biopsies from a patient with NSCLC revealed that the semiquantitative results of the mutation analysis by the SNV panel correlated with the evolution of the disease. CONCLUSIONS: The nCounter platform requires less DNA than NGS and can be employed for routine mutation testing in liquid biopsies of patients with cancer.
Grants:	European Commission 765492 Instituto de Salud Carlos III FIS PI16/00890
Rights:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Language:	Anglès
Document:	Article ; recerca ; Versió publicada
Published in:	Clinical Chemistry, Vol. 67 Núm. 3 (january 2021) , p. 554-563, ISSN 1530-8561

DOI: 10.1093/clinchem/hvaa248
PMID: 33439966

10 p, 654.4 KB

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Research literature > UAB research groups literature > Research Centres and Groups (research output) > Health sciences and biosciences > Institut d'Investigació en Ciencies de la Salut Germans Trias i Pujol (IGTP)
Research literature > UAB research groups literature > Research Centres and Groups (research output) > Health sciences and biosciences > Institut de Recerca Sant Pau
Articles > Research articles
Articles > Published articles