IDENTIFICATION OF 3 NOVEL MUTATIONS IN THE CFTR GENE USING TEMPERATURE-OPTIMIZED NONRADIOACTIVE CONDITIONS FOR SSCP ANALYSIS

Optimal temperature conditions for the detection of 28 known mutations on 15 exons of the human cystic fibrosis transmembrane conductance re gulator gene by single strand conformation polymorphism analysis using the Diagen TGGE Apparatus were established. This procedure was applie d to the detection of unknown mutations in 58 non-deltaF(508) chromoso mes. Three novel mutations, -471del3 (5' flanking region), 3171insC (e xon 17a) and 4700(T)(8/9) (3' non-translated region) of the CFTR gene were found. Mutation 3171insC occured in conjunction with the delta F- 508 mutation on the other allele of a child presenting with severe pat hology. Mutation -471de13 has so far only been found in one healthy in dividual and her father, and 4700(T)(8/9) is a DNA sequence polymorphi sm.