title: A preliminary Quality Control (QC) for next generation sequencing (NGS) library evaluation turns out to be a very useful tool for a rapid detection of BRCA1/2 deleterious mutations.
abstract: BACKGROUND
  Recent advances in next generation sequencing (NGS) technology have enabled comprehensive and accurate screening of the entire genomic region of BRCA1/2 genes and, to date, many studies report the effectiveness of these technologies. Here we show that Gene Scan (GS) labeling Quality Control (QC), performed before massive parallel pyrosequencing, coupled with Multiple Amplicon Quantification software (MAQ-S) analysis is a rapid and powerful tool in the detection of deleterious BRCA mutations carried by different patients.
  
  
  METHODS
  GS labeling QC assay was performed according to the manufacturers' instructions and MAQ-S software was employed for analysis results.
  
  
  RESULTS
  GS labeling QC was able to detect 14 different BRCA frameshift mutations in our patients. In addition, two novel BRCA mutations (c.1893_1894insTTAAGCCCACAAAT in BRCA1 gene and c.9413_9414insT in BRCA2 gene) were identified.
  
  
  CONCLUSIONS
  We prove that a simple QC step may represent a valid and useful tool for a rapid detection of frameshift mutations in BRCA genes. For this reason, we recommend using this approach before massive parallel sequencing.
authors.affiliation: Laboratory of Molecular Biology, Institute of Biochemistry and Clinical Biochemistry, Catholic University of Sacred Heart, Largo A. Gemelli 8, 00168 Rome, Italy. Electronic address: paolaconcolino78@libero.it.
authors.forename: Paola
authors.initials: P
authors.lastname: Concolino
authors.affiliation: Laboratory of Molecular Biology, Institute of Biochemistry and Clinical Biochemistry, Catholic University of Sacred Heart, Largo A. Gemelli 8, 00168 Rome, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Costella
authors.affiliation: Laboratory of Molecular Biology, Institute of Biochemistry and Clinical Biochemistry, Catholic University of Sacred Heart, Largo A. Gemelli 8, 00168 Rome, Italy.
authors.forename: Angelo
authors.initials: A
authors.lastname: Minucci
authors.affiliation: Laboratory of Molecular Biology, Institute of Biochemistry and Clinical Biochemistry, Catholic University of Sacred Heart, Largo A. Gemelli 8, 00168 Rome, Italy.
authors.forename: Giovanni Luca
authors.initials: GL
authors.lastname: Scaglione
authors.affiliation: Laboratory of Molecular Biology, Institute of Biochemistry and Clinical Biochemistry, Catholic University of Sacred Heart, Largo A. Gemelli 8, 00168 Rome, Italy.
authors.forename: Concetta
authors.initials: C
authors.lastname: Santonocito
authors.affiliation: Department of Obstetrics and Gynecology, Catholic University of Sacred Heart, Largo A. Gemelli 8, 00168 Rome, Italy.
authors.forename: Vanda
authors.initials: V
authors.lastname: Salutari
authors.affiliation: Department of Obstetrics and Gynecology, Catholic University of Sacred Heart, Largo A. Gemelli 8, 00168 Rome, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Scambia
authors.affiliation: Laboratory of Molecular Biology, Institute of Biochemistry and Clinical Biochemistry, Catholic University of Sacred Heart, Largo A. Gemelli 8, 00168 Rome, Italy.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Zuppi
authors.affiliation: Laboratory of Molecular Biology, Institute of Biochemistry and Clinical Biochemistry, Catholic University of Sacred Heart, Largo A. Gemelli 8, 00168 Rome, Italy. Electronic address: ecapoluongo@rm.unicatt.it.
authors.forename: Ettore
authors.initials: E
authors.lastname: Capoluongo
mesh_terms: D024682:BRCA2 Protein / Q000235:genetics*; D001483:Base Sequence; D001943:Breast Neoplasms / Q000175:diagnosis / Q000235:genetics; D005260:Female; D015723:Gene Library; D005820:Genetic Testing / Q000592:standards*; D059014:High-Throughput Nucleotide Sequencing / Q000592:standards; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D010051:Ovarian Neoplasms / Q000175:diagnosis / Q000235:genetics; D011786:Quality Control; D017422:Sequence Analysis, DNA / Q000592:standards*; D013997:Time Factors; D044767:Ubiquitin-Protein Ligases / Q000235:genetics*
chemical_list: D024682:BRCA2 Protein; C551750:BRCA2 protein, human; C111164:BRAP protein, human; D044767:Ubiquitin-Protein Ligases
keywords: BRCA1; BRCA2; Breast cancer; Frameshift mutations; NGS