The extended CF is now available for the identification of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene mutations based on polymerase chain reaction (PCR) and reverse-hybridization. For human in vitro diagnostics.
The procedure includes three steps:
(1) DNA isolation
(2) PCR amplification using biotinylated primers
(3) hybridization of amplification products to test strips containing allele-specific oligonucleotide probes immobilized as an array of parallel lines.
Bound biotinylated sequences are detected using streptavidin-alkaline phosphatase and color substrates. The assay covers 38 mutations and 1 polymorphism in the CFTR gene: CFTRdel2,3 (21kb), I507del (-ATC), F508del (-CTT), 1677delTA, 1717-1G>A, G542X, G551D, R553X, 2043delG, 2143delT, 2183AA>G, 2184delA, 2184insA, 2789+5G>A, R1158X, R1162X, 3659delC, 3905insT, W1282X, N1303K [Teststrip A]; E60X, G85E, 394delTT, E92X, E92K, R117H, 621+1G>T, 1078delT, R334W, I336K, R347H, R347P, A455E, 3120+1G>A, 3272-26A>G, Y1092X, M1101K, 3849+10kbC>T, IVS8 5T/7T/9T [Teststrip B].