The authors declare that they have no competing interests.
ME carried out the molecular genetic studies, took part in gathering the patients’ recruitment and drafted the manuscript. BD participated in the analysis of the HRM curve. KH and AB revised the manuscript. MAS helped analyze the sequencing result. AB assisted in the sequencing experiment. HJ and AB coordinated the patient’s recruitment and provided the clinical data. AB supervised the sequencing experiment. SN conceived the study and participated in its design and coordination. All authors read and approved the final manuscript.
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.
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.
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.
Our results strongly suggest that the HRM approach represents a reliable and highly sensitive method for mutation scanning, especially in diagnosis.
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- Screening of exon 11 of BRCA1 gene using the high resolution melting approach for diagnosis in Moroccan breast cancer patients
Meryam El Khachibi
Mohamed Amine Senhaji
- BioMed Central
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