✦ LIBER ✦

Massive parallel amplicon sequencing of the breast cancer genes BRCA1 and BRCA2: opportunities, challenges, and limitations

✍ Scribed by Kim De Leeneer; Jan Hellemans; Joachim De Schrijver; Machteld Baetens; Bruce Poppe; Wim Van Criekinge; Anne De Paepe; Paul Coucke; Kathleen Claes

Publisher: John Wiley and Sons
Year: 2011
Tongue: English
Weight: 584 KB
Volume: 32
Category: Article
ISSN: 1059-7794
DOI: 10.1002/humu.21428

No coin nor oath required. For personal study only.

✦ Synopsis

This study describes how the new massive parallel sequencing technology can be implemented in a diagnostic setting for the breast cancer susceptibility genes (BRCA1 and BRCA2). The throughput was maximized by increasing uniformity in coverage, obtained by a multiplex approach, which outperformed pooling of singleplex PCRs. We evaluated the sensitivity by analysis of 133 distinct sequence variants; three (2%) deletions or duplications in homopolymers of greater than or equal to seven nucleotides remained undetected, illustrating a limitation of pyrosequencing. Furthermore, other limitations like nonrandom sequencing errors, pseudogene amplification, and failure to detect multiexon deletions are thoroughly described. Our workflow illustrates the potential of massive parallel sequencing of large genes in a diagnostic setting, which is of great importance to meet the increasing expectations of genetic testing. Implementation of this approach will hopefully lead to a strong reduction in turnaround times. As a consequence a wider spectrum of at risk women will be able to benefit from therapeutic interventions and prophylactic interventions.

📜 SIMILAR VOLUMES

Germline mutations in the BRCA1 and BRCA

Germline mutations in the BRCA1 and BRCA2 genes in Turkish breast/ovarian cancer patients

✍ A. Esra Manguoǧlu; Güven Lüleci; Tayfun Özçelik; Taner Çolak; Hagit Schayek; Mus 📂 Article 📅 2003 🏛 John Wiley and Sons 🌐 English ⚖ 47 KB 👁 2 views

In this study we genotyped Turkish breast/ovarian cancer patients for BRCA1/BRCA2 mutations: protein truncation test (PTT) for exon 11 BRCA1 of and, multiplex PCR and denaturing gradient gel electrophoresis (DGGE) for BRCA2, complemented by DNA sequencing. In addition, a modified restriction assay w

Novel germline mutations in the BRCA1 an

Novel germline mutations in the BRCA1 and BRCA2 genes in Indian breast and breast-ovarian cancer families

✍ Mani T. Valarmathi; Meenakshi Sawhney; Suryanarayana S. V. Deo; Nootan K. Shukla 📂 Article 📅 2004 🏛 John Wiley and Sons 🌐 English ⚖ 97 KB 👁 2 views

The two major hereditary breast/ovarian cancer predisposition tumor suppressor genes, BRCA1 and BRCA2 that perform apparently generic cellular functions nonetheless cause tissue-specific syndromes in the human population when they are altered, or mutated in the germline. However, little is known abo

Analysis of breast cancer susceptibility

Analysis of breast cancer susceptibility genes BRCA1 and BRCA2 in Thai familial and isolated early-onset breast and ovarian cancer

✍ Pimpicha Patmasiriwat; Kris Bhothisuwan; Olga M. Sinilnikova; Sandrine Chopin; S 📂 Article 📅 2002 🏛 John Wiley and Sons 🌐 English ⚖ 173 KB 👁 2 views

Here we report the study on BRCA1 and BRCA2 mutations in 12 Thai breast and/or ovarian cancer families and 6 early-onset breast or breast/ovarian cancer cases without a family history of cancer. Five distinct rare alterations were identified in each gene: four introducing premature stop codons, one

Breast cancer risk in BRCA1 and BRCA2 mu

Breast cancer risk in BRCA1 and BRCA2 mutation carriers and polyglutamine repeat length in the AIB1 gene

✍ David J. Hughes; Sophie M. Ginolhac; Isabelle Coupier; Laure Barjhoux; Valérie G 📂 Article 📅 2005 🏛 John Wiley and Sons 🌐 French ⚖ 89 KB 👁 2 views

## Abstract Marked variation in phenotypic expression among __BRCA1__ and __BRCA2__ mutation carriers may be partly explained by modifier genes that influence mutation penetrance. Variation in CAG/CAA repeat lengths coding for stretches of glutamines in the C‐terminus of the AIB1 protein (amplified

Germline mutations of the BRCA1-associat

Germline mutations of the BRCA1-associated ring domain (BARD1) gene in breast and breast/ovarian families negative for BRCA1 and BRCA2 alterations

✍ Chiara Ghimenti; Elisa Sensi; Silvano Presciuttini; Isa Maura Brunetti; PierFran 📂 Article 📅 2002 🏛 John Wiley and Sons 🌐 English ⚖ 188 KB 👁 3 views

## Abstract BARD1 (BRCA1‐associated RING domain) was identified by yeast two‐hybrid screening as a protein interacting with BRCA1. Somatic and germline mutations of __BARD1__ have been detected in sporadic breast, ovarian, and endometrial cancers. The present study represents the first description

High frequency of recurrent mutations in

High frequency of recurrent mutations in BRCA1 and BRCA2 genes in Polish families with breast and ovarian cancer

✍ Ewa Grzybowska; Helena Zientek; Anna Jasinska; Marek Rusin; Piotr Kozlowski; Krz 📂 Article 📅 2000 🏛 John Wiley and Sons 🌐 English ⚖ 347 KB 👁 2 views