More information on how to interpret observations on these control fragments can be found in the MLPA General Protocol and online at SALSA Artificial Duplication DNA SD024: In case no positive DNA sample is available in your laboratory, an artificial duplication DNA sample for this probemix (catalogue number SD024) can be ordered from MRC Holland. 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. Complete probe sequences and the identity of the genes detected by the reference probes are available online. In addition, ten reference probes are included that detect autosomal chromosomal locations. One probe is included for exon 1b, an alternative first exon that is not present in BRCA1 transcript variant 1, and two probes detect sequences located 4.7 kb and 0.8 kb upstream of the BRCA1 gene. Three probes are present for exon 24 and two probes for exon 16. Three probes are present for exon 13, which is frequently deleted or duplicated (Hogervorst et al. Eight probes are present for exon 11 (3426 nt long). This includes 38 probes for the BRCA1 gene region.Īt least one MLPA probe is present for each exon in the major BRCA1 transcript variant 1. Probemix content: The SALSA MLPA probemix P002-D1 BRCA1 contains 48 MLPA probes with amplification products between 130 and 469 nucleotides (nt). 1997), while in a Danish cohort of HBOC patients the prevalence was 3.8% (Thomassen et al. 2003), in the Netherlands 27%-36% (Hogervorst et al. For example in Italian HBOC families the prevalence is 23% (Montagna et al. Large genomic rearrangements (LGRs) in BRCA1 may account for up to 25% of all disease-causing mutations, dependent on the population (Smith et al. These copy number changes are usually missed by amplicon-based sequencing analysis (Sanger sequencing or Next Generation Sequencing), but can be detected by the MLPA technique and hence MLPA complements sequence analysis of the BRCA1 gene. Deletions and duplications of complete exons in the BRCA1 gene are the second most common cause of defects in the BRCA1 gene. The great majority of germline defects in the BRCA1 gene are point mutations that can be detected by sequence analysis. In addition, mutations in the BRCA1 and BRCA2 genes account for around 15% of ovarian cancers overall. Mutations in the BRCA1 and BRCA2 genes account for about 20 to 25% of hereditary breast cancers and about 5 to 10% of all breast cancers. Features characteristic of hereditary, versus sporadic, breast cancer are: younger age at diagnosis, frequent bilateral disease, and more frequent occurrence of disease among male relatives. Clinical background: Germline defects in the BRCA1 gene are the most frequent cause of a hereditary predisposition to breast cancer. 2 To be used in combination with a SALSA MLPA Reagent Kit and Coffalyser.Net analysis software. In all other countries, the product is for research use only (RUO). 1 Please note that this probemix is for in vitro diagnostic (IVD) use in the countries specified at the end of the product description. ![]() ![]() This device is not intended to be used for standalone diagnostic purposes, pre-implantation or prenatal testing, population screening, or for the detection of, or screening for, acquired or somatic genetic aberrations, e.g from DNA extracted from formalin-fixed paraffin embedded (FFPE) or fresh tumour materials. ![]() The results of this test should be interpreted by a clinical molecular geneticist or equivalent. It is therefore recommended to use this SALSA MLPA probemix in combination with sequence analysis.Īssay results are intended to be used in conjunction with other clinical and diagnostic findings, consistent with professional standards of practice, including confirmation by alternative methods, clinical genetic evaluation, and counselling, as appropriate. Most defects in the BRCA1 gene are point mutations, none of which will be detected by MLPA. In particular, CNVs detected by only a single probe always require confirmation by another method. P002 BRCA1 is intended to confirm a potential cause for and clinical diagnosis of hereditary breast and ovarian cancer (HBOC) syndrome and for molecular genetic testing of at-risk family members.Ĭopy number variations (CNVs) detected with the P002 BRCA1 probemix should be confirmed with the SALSA MLPA P087 BRCA1 Confirmation probemix or a different technique. Intended purpose: The SALSA MLPA Probemix P002 BRCA1 is an in vitro diagnostic (IVD) 1 or research use only (RUO) semi-quantitative assay 2 for the detection of deletions or duplications in the human BRCA1 gene in genomic DNA isolated from human peripheral whole blood specimens.
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