ORIGINAL ARTICLE

A novel BRCA2 mutation in an Indonesian family found with a new, rapid, and sensitive mutation detection method based on pooled denaturing gradient gel electrophoresis and targeted sequencing

D Purnomosari¹, D K Paramita¹, T Aryandono², G Pals³ and P J van Diest⁴

¹ Department of Histology and Cell Biology, Gadjah Mada University, Jogjakarta 55281, Indonesia
² Department of Surgery, Faculty of Medicine, Gadjah Mada University, Jogjakarta, Indonesia
³ Department of Clinical Genetics, University Medical Centre Utrecht, PO Box 85500, 3508 GA, The Netherlands
⁴ Department of Pathology, University Medical Centre Utrecht

Correspondence to:
Correspondence to:
Professor P J van Diest
Department of Pathology, University Medical Centre Utrecht, PO Box 85500, 3508 GA Utrecht, The Netherlands; p.j.vandiest{at}azu.nl

Background: Breast cancer is increasing in Indonesia and other developing countries. Germline mutations in the BRCA1/2 genes are most strongly associated with a high risk for breast cancer development. There have been no reports on BRCA1/2 gene mutations in the Indonesian population. Genetic research yielding insight into mutations affecting the Indonesian population can help in risk assessment of individual patients.

Aims: To screen the BRCA1/2 genes for mutations in early onset Indonesian breast cancer patients and their families with a new, simple, and sensitive BRCA1/2 mutation screening strategy based on denaturing gradient gel electrophoresis (DGGE) and targeted sequencing.

Methods: DNA was isolated from the blood of four Indonesian breast cancer patients from high risk families and seven family members, and the polymerase chain reaction was performed with specially designed primers throughout the BRCA1/2 coding sequences to produce fragments suitable for pooled DGGE analysis. The aberrantly migrating samples were reamplified and sequenced.

Results: Two mutations were found in exons 13 and 16 of BRCA1 and two mutations in exons 2 and 14 of BRCA2, which turned out to be established polymorphisms according to the Breast Cancer Information Core. In addition, a novel 6 bp deletion in exon 11, leading to a premature stop, was found in BRCA2.

Conclusion: Pooled DGGE and targeted sequencing revealed four BRCA1/2 polymorphisms and one novel BRCA2 mutation in a group of Indonesian patients at high risk of hereditary breast cancer. This illustrates that the proposed method is sensitive and particularly suited for screening unknown populations.

Abbreviations: DGGE, denaturing gradient gel electrophoresis; PCR, polymerase chain reaction; PTT, protein truncation test; SCCP, single strand conformational polymorphism

Keywords: BRCAA1; BRCA2; hereditary breast cancer; methodology; mutation detection

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This article has been cited by other articles:

• van Diest, P J, Pals, G (2007). A novel rapid and sensitive BRCA1/2 mutation detection method based on pooled DGGE and targeted sequencing. J. Clin. Pathol. 60: 478-478 [Full Text]