General information
The SALSA MS-MLPA Probemix ME053-A1 BRCA1-BRCA2-RAD51C is a research use only (RUO) assay for the detection of methylation status of one or more sequences of the BRCA1, BRCA2 and RAD51C genes.

BRCA1, BRCA2 and RAD51C are tumour suppressor genes that participate in DNA repair processes and genome integrity maintenance. Germline deleterious variants in these genes have been extensively described in hereditary breast and ovarian cancer patients, contributing to increased risks of developing the disease. Previous studies have also identified aberrant promoter methylation levels in BRCA1 and RAD51C genes, proposing a new mechanism of gene inactivation in early-onset breast and/or ovarian cancer (Hansmann et al. 2012, Evans et al. 2018). Furthermore, somatic methylation of BRCA1 and BRCA2 may result in reduced gene expression levels and contribute to tumorigenesis and development of sporadic breast/ovarian cancer (Esteller et al. 2000, Bosviel et al. 2011, Stordal et al. 2013). Moreover, promoter hypermethylation of BRCA1, BRCA2 and RAD51C genes has been described in triple-negative breast cancer and epithelial ovarian carcinomas, and it can guide effective therapeutic interventions with PARP inhibitors and serve as markers of therapy response (Veeck et al. 2010, Cunningham et al. 2014, Pennington et al. 2014, Sahnane et al. 2020, Menghi et al. 2022).

This SALSA MS-MLPA probemix is not CE/FDA registered for use in diagnostic procedures. Purchase of this product includes a limited license for research purposes.

Probemix content
The SALSA MS-MLPA Probemix ME053-A1 BRCA1-BRCA2-RAD51C contains 38 (MS-)MLPA probes with amplification products between 123 and 400 nucleotides (nt). Ten MS-MLPA probes contain an HhaI recognition site and provide information on the methylation status of selected GCGC sites in the promoter regions of BRCA1, BRCA2 and RAD51C genes. All probes present will also give information on copy number changes in the analysed sample. This probemix includes also 11 copy number probes flanking BRCA1, BRCA2 and RAD51C genes. In addition, 13 reference probes are included that are not affected by HhaI digestion and target relatively copy number stable regions in various cancer types including breast and ovarian cancer. Also, three digestion control probes are included in this probemix indicating whether or not restriction endonuclease digestion in the MS-MLPA reaction was complete. Complete/partial probe sequences and the identity of the genes detected by the reference probes are available in Table 2 of the Product Description and online (www.mrcholland.com).

This probemix contains nine quality control fragments generating amplification products between 64 and 105 nt: four DNA Quantity fragments (Q-fragments), two DNA Denaturation fragments (D-fragments), one Benchmark fragment, and one chromosome X and one chromosome Y-specific fragment. More information on how to interpret observations on these control fragments can be found in the MS-MLPA General Protocol and online at www.mrcholland.com.