TY - JOUR
T1 - Is quantitative real time polymerase chain reaction MCAM transcript assay really suitable for prognostic and predictive management of melanoma patients?
AU - Capoluongo, Ettore Domenico
AU - Paolillo, Carmela
AU - Vendittelli, Francesca
PY - 2014
Y1 - 2014
N2 - 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_1894in5TTAAGCCCACAAAT in BRCA1 gene and c.9413_9414insT in BRCA2 gene) were identified.
Conclusion: 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. (C) 2014 Elsevier B.V. All rights reserved.
AB - 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_1894in5TTAAGCCCACAAAT in BRCA1 gene and c.9413_9414insT in BRCA2 gene) were identified.
Conclusion: 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. (C) 2014 Elsevier B.V. All rights reserved.
KW - BRCA1-2
KW - Frameshift mutations
KW - BRCA1-2
KW - Frameshift mutations
UR - http://hdl.handle.net/10807/64026
U2 - 10.1111/bjd.12818
DO - 10.1111/bjd.12818
M3 - Article
SN - 1365-2133
VL - 171
SP - 190
EP - 191
JO - British Journal of Dermatology
JF - British Journal of Dermatology
ER -