Identification of Novel FBN1 Mutations in Patients with Marfan Syndrome using DHPLC Analysis.

Identification of Novel FBN1 Mutations in Patients with Marfan Syndrome using DHPLC Analysis. M. Grasso 1, S. Ansaldi 1, A. Mori 1, A. Pisani 1, L. Lanzarini 2, A. Pilotto 1, C. Lucchelli 1, L. Tavazzi 2, E. Arbustini 1; 1 Cardiovascular Pathology and Molecular Diagnostic Lab, Transplant Research Area, IRCCS Policlinico San Matteont, Pavia, Italy, 2 Cardiology Division, IRCCS Policlinico San Matteo, Pavia, Italy. Marfan Syndrome (MFS, MIM#154700) is an autosomal dominant inherited connective tissue disorder (prevalence:1/5000) caused by mutations in the fibrillin-1 gene (FBN1, 15q21). The disorder is characterised by highly variable phenotypic manifestations, mainly in cardiovascular, ocular and skeletal systems. The FBN1 (230 Kb, 65 exons, 2871 amino acids) has revealed more than 500 mutations. We describe 11 novel mutations that were identified in 12 probands (one with sporadic and ten with familial disease). The MFS diagnosis was evaluated following the revised diagnostic criteria of the Ghent nosology. The FBN1 gene was analysed using DHPLC technology (Transgenomic) and automated sequencing (ABI 3100).All family members were tested for the mutations found. These mutations were absent in 50 controls. Our results suggest that DHPLC is a reliable and cost-effective technique for the screening of such a large gene and that FBN1 screening could be a helpful tool to confirm and possibly anticipate the clinical diagnosis in familial cases.