PheoSeq: A Targeted Next-Generation Sequencing Assay for Pheochromocytoma and Paraganglioma Diagnostics

Maria Currás-Freixes, Elena Piñeiro-Yañez, Cristina Montero-Conde, María Apellániz-Ruiz, Bruna Calsina, Veronika Mancikova, Laura Remacha, Susan Richter, Tonino Ercolino, Natalie Rogowski-Lehmann, Timo Deutschbein, María Calatayud, Sonsoles Guadalix, Cristina Álvarez-Escolá, Cristina Lamas, Javier Aller, Julia Sastre-Marcos, Conxi Lázaro, Juan C. Galofré, Ana Patiño-GarcíaAmparo Meoro-Avilés, Judith Balmaña-Gelpi, Paz De Miguel-Novoa, Milagros Balbín, Xavier Matías-Guiu, Rocío Letón, Lucía Inglada-Pérez, Rafael Torres-Pérez, Juan M. Roldán-Romero, Cristina Rodríguez-Antona, Stephanie M.J. Fliedner, Giuseppe Opocher, Karel Pacak, Esther Korpershoek, Ronald R. de Krijger, Laurent Vroonen, Massimo Mannelli, Martin Fassnacht, Felix Beuschlein, Graeme Eisenhofer, Alberto Cascón, Fátima Al-Shahrour, Mercedes Robledo

Onderzoeksoutput: Bijdrage aan tijdschriftArtikelpeer review

63 Citaten (Scopus)

Samenvatting

Genetic diagnosis is recommended for all pheochromocytoma and paraganglioma (PPGL) cases, as driver mutations are identified in approximately 80% of the cases. As the list of related genes expands, genetic diagnosis becomes more time-consuming, and targeted next-generation sequencing (NGS) has emerged as a cost-effective tool. This study aimed to optimize targeted NGS in PPGL genetic diagnostics. A workflow based on two customized targeted NGS assays was validated to study the 18 main PPGL genes in germline and frozen tumor DNA, with one of them specifically directed toward formalin-fixed paraffin-embedded tissue. The series involved 453 unrelated PPGL patients, of whom 30 had known mutations and were used as controls. Partial screening using Sanger had been performed in 275 patients. NGS results were complemented with the study of gross deletions. NGS assay showed a sensitivity ≥99.4%, regardless of DNA source. We identified 45 variants of unknown significance and 89 pathogenic mutations, the latter being germline in 29 (7.2%) and somatic in 58 (31.7%) of the 183 tumors studied. In 37 patients previously studied by Sanger sequencing, the causal mutation could be identified. We demonstrated that both assays are an efficient and accurate alternative to conventional sequencing. Their application facilitates the study of minor PPGL genes, and enables genetic diagnoses in patients with incongruent or missing clinical data, who would otherwise be missed.

Originele taal-2Engels
Pagina's (van-tot)575-588
Aantal pagina's14
TijdschriftJournal of Molecular Diagnostics
Volume19
Nummer van het tijdschrift4
DOI's
StatusGepubliceerd - jul. 2017
Extern gepubliceerdJa

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