A RAPID, EFFICIENT, AND SENSITIVE ASSAY FOR SIMULTANEOUS DETECTION OFMULTIPLE CYSTIC-FIBROSIS MUTATIONS

We describe the use of DGGE multiplex systems for rapid analysis of 15 exons of the cystic fibrosis transmembrane conductance regulator (CFT R) gene, in which about half of the known CF molecular defects are clu stered. We have previously determined the spectrum of mutations affect ing the CFTR gene in the French population using a strategy based on d enaturing gradient gel electrophoresis (DGGE) of amplified gene segmen ts. Analysis of CF patients' DNA with five DGGE multiplex systems perm itted us to characterize nearly 35% of non-DELTAF508 CF alleles and in creased the CF allele detection rate to almost 82% in this population. This simple and rapid multiplex analysis strategy, which allows a sig nificant proportion of the most frequent CF mutations in Caucasians to be detected, will be helpful in the implementation of genetic screeni ng programs. (C) 1993 Wiley-Liss, Inc.