Simultaneous screening for 11 mutations in the cystic fibrosis transmembrane conductance regulator gene by multiplex amplification and reverse dot-blot

摘要: An assay is described in which 11 mutations the cystic fibrosis transmembrane conductance regulator (CFTR) gene can be screened simultaneously. Six different exons of CFTR are amplified a single multiplex amplification. Biotinylated dUTP incorporated into fragments during amplification process. A sample this mixture then hybridized to 21 poly-dT tailed oligonucleotide probes bound nylon membrane. In order screen step hybridization, length oligonucleotides and amount used were optimized. The detection performed by binding avidin-alkaline phosphatase biotin, followed chemiluminescent reaction. By means fast sensitive assay, about 85% all Belgian population detected.