A single-tube multiplex system for the simultaneous detection of 10 common cystic fibrosis mutations

We describe a nonisotopic single-tube polymerase chain reaction (PCR) multiplex system that detects 10 of the more common cystic fibrosis (CF) mutations (delta F508, delta I507, V520F, G551D, G542X, R553X, R117H, 621 + 1G-->T, N1303K, A455E). The use of this method detects approximately 90% of the CF alleles in the British population. Five exons of the CF gene are amplified simultaneously by PCR, followed by an overnight triple enzyme restriction digest, and then resolved by high-resolution gel acrylamide electrophoresis.