NF1 MUTATION ANALYSIS USING A COMBINED HETERODUPLEX SSCP APPROACH/

Neurofibromatosis type 1 (NF1) is a common autosomal dominant disorder characterized predominantly by neurofibromas, cafe-au-lait spots, and Lisch nodules. The disease is caused by disruptive mutations of the l arge NF1 gene, with half of cases caused by new mutation. Less than 10 0 constitutional mutations have thus far been published, ranging from very large deletions to point mutations. We have pursued NF1 mutation analysis by heteroduplex analysis (HDA) and single strand conformation al polymorphism analysis (SSCP) of individual exons, We streamlined th ese techniques to eliminate the use of radioactivity, to apply both me thods to the same PCR product, and to multiplex samples in gels. Appli ed simultaneously to a set of 67 unrelated NF1 patients, HDA and SSCP have thus far identified 26 mutations and/or variants in 45 of the 59 exons tested. Disease causing mutations were found in 19% (13/67) of c ases studied. Both techniques detected a variety of mutations includin g splice mutations, insertions, deletions, and point changes, with som e overlap in the ability of each method to detect variants. (C) 1997 W iley-Liss, Inc.