: The SALSA MLPA probemix P091 CFTR is an in vitro diagnostic (IVD)1
or research use only (RUO) assay for the detection of the ∆F508 and ∆I507 mutations, deletions and/or duplications of one or more exons in the human CFTR
gene of patients diagnosed with cystic fibrosis (CF) or congenital absence of the vas deferens (CAVD). When an aberration has been identified in patient DNA, this product can also be used for carrier screening of family members. This assay is optimised for use with peripheral blood derived genomic DNA.
Deletions or duplications obtained with the P091 CFTR probemix must be verified by another technique. In particular, copy number changes detected by only a single probe always require validation by another method. This SALSA MLPA probemix is not intended to be used as a standalone assay for clinical decisions. Most defects in the CFTR
gene are point mutations. Except ∆F508 and ∆I507, the majority of point mutations will not be detected by MLPA. It is therefore recommended to use this SALSA MLPA probemix in combination with sequence analysis. The results of this test should be interpreted by a clinical molecular geneticist or equivalent.
Please note that this probemix is for In Vitro Diagnostic use (IVD) in the countries specified at the end of this product description. In all other countries, the product is for Research Use Only (RUO).
: Cystic fibrosis (CF) and congenital absence of the vas deferens (CAVD), which are both recessive genetic diseases, are caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR
) gene. The CFTR gene product functions as a chloride channel regulating water and ion transport across membranes in the lungs, liver, pancreas, intestine, reproductive tracts, and sweat glands. Azoospermia can occur in men with CAVD. CF related pathologies include thickened mucus in the lungs with frequent respiratory infection and inflammation, and malnutrition and diabetes due to pancreatic insufficiency. More information is available on www.ncbi.nlm.nih.gov/books/NBK1250
Deletions and duplications of complete exons in the CFTR
gene account for 1-2% of mutations found in patients and are usually missed by standard sequence analysis (Svensson et al. 2010). Most of these deletions and duplications can be detected by the MLPA technique and hence MLPA complements sequence analysis of the CFTR
gene. By far the most common mutation found in patients is DF508 (allele frequency of 65-70% (Bobadilla et al. 2002)). The wild type allele of this mutation can be detected with this probemix.
: This SALSA®
probemix P091 CFTR contains 44 MLPA probes with amplification products between 130 nt and 481 nt including 34 probes for the CFTR gene region and 10 reference probes that detect sequences outside this region. The identity of the genes detected by the reference probes is available online (www.mlpa.com
The probemix contains probes for each of the 27 CFTR
exons and second probes for exons 1, 11, 13 and 27. One of the exon 11 (330 nt) probes detects the wild type allele of the ∆F508 mutation. A reduced signal for this probe can point towards the presence of the ∆F508 mutation, the ∆I507 mutation, or a deletion of exon 11.
This probemix contains nine quality control fragments generating amplification products between 64 nt and 121 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 MLPA General Protocol and online at www.mlpa.com