PheoSeq

Autor: Ana Patiño-García, Martin Fassnacht, Massimo Mannelli, Alberto Cascón, Conxi Lázaro, María Calatayud, Cristina Álvarez-Escolá, Graeme Eisenhofer, Judith Balmaña-Gelpi, Karel Pacak, Felix Beuschlein, Maria Currás-Freixes, Mercedes Robledo, Laurent Vroonen, Rafael Torres-Pérez, Susan Richter, Cristina Lamas, Bruna Calsina, Xavier Matias-Guiu, Amparo Meoro-Avilés, Timo Deutschbein, Lucía Inglada-Pérez, Milagros Balbín, Ronald R. de Krijger, Javier Aller, Julia Sastre-Marcos, Juan María Roldan-Romero, Stephanie M. J. Fliedner, Natalie Rogowski-Lehmann, Rocío Letón, Juan C. Galofré, Sonsoles Guadalix, María Apellániz-Ruiz, Paz de Miguel-Novoa, Tonino Ercolino, Laura Remacha, Fatima Al-Shahrour, Giuseppe Opocher, Elena Piñeiro-Yáñez, Veronika Mancikova, Cristina Montero-Conde, Esther Korpershoek, Cristina Rodríguez-Antona
Rok vydání: 2017
Předmět:
Zdroj: The Journal of Molecular Diagnostics. 19:575-588
ISSN: 1525-1578
DOI: 10.1016/j.jmoldx.2017.04.009
Popis: 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.
Databáze: OpenAIRE
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