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High-resolution melting (HRM) assay for the detection of recurrent BRCA1/BRCA2 germline mutations in Tunisian breast/ovarian cancer families

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Abstract

Germline deleterious mutations in the BRCA1/BRCA2 genes are associated with an increased risk for the development of breast and ovarian cancer. Given the large size of these genes the detection of such mutations represents a considerable technical challenge. Therefore, the development of cost-effective and rapid methods to identify these mutations became a necessity. High resolution melting analysis (HRM) is a rapid and efficient technique extensively employed as high-throughput mutation scanning method. The purpose of our study was to assess the specificity and sensitivity of HRM for BRCA1 and BRCA2 genes scanning. As a first step we estimate the ability of HRM for detection mutations in a set of 21 heterozygous samples harboring 8 different known BRCA1/BRCA2 variations, all samples had been preliminarily investigated by direct sequencing, and then we performed a blinded analysis by HRM in a set of 68 further sporadic samples of unknown genotype. All tested heterozygous BRCA1/BRCA2 variants were easily identified. However the HRM assay revealed further alteration that we initially had not searched (one unclassified variant). Furthermore, sequencing confirmed all the HRM detected mutations in the set of unknown samples, including homozygous changes, indicating that in this cohort, with the optimized assays, the mutations detections sensitivity and specificity were 100 %. HRM is a simple, rapid and efficient scanning method for known and unknown BRCA1/BRCA2 germline mutations. Consequently the method will allow for the economical screening of recurrent mutations in Tunisian population.

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Acknowledgments

The authors thank all the patients and the clinicians at the Salah azaiz Cancer Institute for taking part in this study. This work was supported by the Ministry of High Education and Scientific Research and Technology.

Conflict of interest

The authors declare that there are no conflicts of interest.

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Authors and Affiliations

  1. Laboratoire Génétique Humaine, Faculté de Médecine de Tunis, University Tunis El Manar, Tunis, Tunisia

    Aouatef Riahi, Maher Kharrat, Imen Lariani & Habiba Chaabouni-Bouhamed

  2. Department of Hereditary and Congenital Disorders, Charles Nicolle Hospital, Bd du 9 Avril, 1006, Tunis, Tunisia

    Habiba Chaabouni-Bouhamed

Authors
  1. Aouatef Riahi
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  2. Maher Kharrat
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  3. Imen Lariani
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  4. Habiba Chaabouni-Bouhamed
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Correspondence to Habiba Chaabouni-Bouhamed.

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Riahi, A., Kharrat, M., Lariani, I. et al. High-resolution melting (HRM) assay for the detection of recurrent BRCA1/BRCA2 germline mutations in Tunisian breast/ovarian cancer families. Familial Cancer 13, 603–609 (2014). https://doi.org/10.1007/s10689-014-9740-5

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  • DOI: https://doi.org/10.1007/s10689-014-9740-5

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