title: BRCA1-2 diagnostic workflow from next-generation sequencing technologies to variant identification and final report.
abstract: The BRCA1-BRCA2 genes predispose to hereditary breast and ovarian cancer, and the germline and mutational status of these genes defines a target population that can benefit from PARP inhibitor treatments. To respond to the increasing number of BRCA1-BRCA2 tests, it is necessary to shift to high-throughput technologies that are reliable and less time consuming. Different methodological platforms are dedicated to this purpose with different approaches and algorithms for analysis. Our aim was to set up a cost-effective and low time-consuming BRCA1-BRCA2 mutation detection workflow using the Ion Torrent PGM technology. A retrospective cohort of 40 patients with familial breast/ovarian cancer previously tested by Sanger sequencing and a prospective cohort of 72 patients (validation set) were analyzed. The validation set included 64 patients affected by familial breast/ovarian cancer and eight sporadic ovarian cancer cases, who are potential candidates for PARPi treatments. A complete and standardized workflow easily usable and suitable in a certified laboratory has been proved and validated. This includes all steps from library preparation to the final report. The use of next-generation sequencing will be of benefit for patients enrolled in the genetic counseling process and, moreover, will enhance the process of selecting patients eligible for personalized treatments. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Molecular Genetics Laboratory, IRCCS Istituto Tumori "Giovanni Paolo II,", Bari, Italy.
authors.forename: Brunella
authors.initials: B
authors.lastname: Pilato
authors.affiliation: Molecular Genetics Laboratory, IRCCS Istituto Tumori "Giovanni Paolo II,", Bari, Italy.
authors.forename: Rosamaria
authors.initials: R
authors.lastname: Pinto
authors.affiliation: Molecular Genetics Laboratory, IRCCS Istituto Tumori "Giovanni Paolo II,", Bari, Italy.
authors.forename: Simona
authors.initials: S
authors.lastname: De Summa
authors.affiliation: Molecular Genetics Laboratory, IRCCS Istituto Tumori "Giovanni Paolo II,", Bari, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Petriella
authors.affiliation: Molecular Genetics Laboratory, IRCCS Istituto Tumori "Giovanni Paolo II,", Bari, Italy.
authors.forename: Rosanna
authors.initials: R
authors.lastname: Lacalamita
authors.affiliation: Molecular Genetics Laboratory, IRCCS Istituto Tumori "Giovanni Paolo II,", Bari, Italy.
authors.forename: Katia
authors.initials: K
authors.lastname: Danza
authors.affiliation: Experimental Medical Oncology Unit, IRCCS Istituto Tumori "Giovanni Paolo II,", Bari, Italy.
authors.forename: Angelo
authors.initials: A
authors.lastname: Paradiso
authors.affiliation: Molecular Genetics Laboratory, IRCCS Istituto Tumori "Giovanni Paolo II,", Bari, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Tommasi
mesh_terms: D019313:BRCA1 Protein / Q000235:genetics*; D024682:BRCA2 Protein / Q000235:genetics*; D001943:Breast Neoplasms / Q000175:diagnosis / Q000235:genetics* / Q000473:pathology; D056915:DNA Copy Number Variations / Q000235:genetics; D005260:Female; D020022:Genetic Predisposition to Disease; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D009154:Mutation; D010051:Ovarian Neoplasms / Q000175:diagnosis / Q000235:genetics* / Q000473:pathology; D057188:Workflow
chemical_list: D019313:BRCA1 Protein; C492913:BRCA1 protein, human; D024682:BRCA2 Protein; C551750:BRCA2 protein, human