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BMC Cancer. 2015 Feb 25;15:81. doi: 10.1186/s12885-015-1040-4.

Screening of exon 11 of BRCA1 gene using the high resolution melting approach for diagnosis in Moroccan breast cancer patients.

Author information

1
Genetics and Molecular Pathology Laboratory, Medical school of Casablanca, Casablanca, Morocco. el.khachibi.meryam@gmail.com.
2
Genetics and Molecular Pathology Laboratory, Medical school of Casablanca, Casablanca, Morocco. br.diakite@yahoo.fr.
3
Genetics and Molecular Pathology Laboratory, Medical school of Casablanca, Casablanca, Morocco. Khalilhamzi@gmail.com.
4
Genetics and Molecular Pathology Laboratory, Medical school of Casablanca, Casablanca, Morocco. abdallahbadou@yahoo.com.
5
Genetics and Molecular Pathology Laboratory, Medical school of Casablanca, Casablanca, Morocco. m.amine.senhaji@gmail.com.
6
Laboratoire de Génétique Moléculaire Humaine, Département de la Recherche Scientifique, Institut Pasteur du Maroc, Casablanca, Morocco. amina-bakhchane@yahoo.fr.
7
Department of Oncology, Ibn Rochd University Hospital, Casablanca, Morocco. h.jouhadi@yahoo.fr.
8
Laboratoire de Génétique Moléculaire Humaine, Département de la Recherche Scientifique, Institut Pasteur du Maroc, Casablanca, Morocco. hamidbarakat@pasteur.ma.
9
Department of Oncology, Ibn Rochd University Hospital, Casablanca, Morocco. beniderabdel@yahoo.fr.
10
Genetics and Molecular Pathology Laboratory, Medical school of Casablanca, Casablanca, Morocco. nadifisel@yahoo.fr.

Abstract

BACKGROUND:

Identification of specific mutations in cancer patients may lead to the discovery of genes, which can affect susceptibility and/or prognosis. It has previously been reported that mutations in BRCA1 and BRCA2 genes are linked to breast cancer. Here, we evaluated the use of the High Resolution Melting (HRM) approach to screen for mutations in exon 11 of BRCA1 gene in Moroccan patients.

METHODS:

HRM analysis was used to screen exon 11 from 71 breast cancer patients in order to detect different variants. Conventional Sanger sequencing was used to confirm the presence of possible mutations. Distribution of different SNPs was determined by SNaPshot analysis software.

RESULTS:

In order to assess the efficacy of the HRM approach to screen for mutations, especially in diagnosis, we first used two samples with previously known mutations, "2924delA and 3398delC". Indeed, these previously known sequence variants were detected by the HRM approach and yielded melting curves with atypical shape relative to wild-type control sequences. We then analyzed, 69 samples from breast cancer patients using the HRM method, and were able to detect two samples with atypical curves. Sequencing of the two samples, using the conventional Sanger approach, confirmed the presence of the same SNP (c.2612C > T) in both samples.

CONCLUSIONS:

Our results strongly suggest that the HRM approach represents a reliable and highly sensitive method for mutation scanning, especially in diagnosis.

PMID:
25885115
PMCID:
PMC4351675
DOI:
10.1186/s12885-015-1040-4
[Indexed for MEDLINE]
Free PMC Article

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