title: A combined analytical approach reveals novel EXT1/2 gene mutations in a large cohort of Italian multiple osteochondromas patients.
abstract: Multiple osteochondromas (MO), also known as hereditary multiple exostoses (HME), is one of the most common hereditary musculoskeletal diseases in Caucasians (1/50,000) with wide clinical variability and genetic heterogeneity. Two genes have thus far been identified as causing the disease, namely EXT1 and EXT2. Various methods to detect mutations in the EXT genes have been used. Here a cohort of 100 MO patients belonging to unrelated Italian families have been analyzed by single-strand conformation polymorphism (SSCP) analysis or by denaturing high performance liquid chromatography (DHPLC). However, neither of these techniques can detect deletions or duplications of entire exons. Families that were negative at SSCP/DHPLC analysis underwent two-color multiple ligation-dependent probe amplification (MLPA) analysis. By these complementary techniques mutation detection was significantly improved and 26 novel mutations have been revealed as well as 18 previously described mutations to give a total of 44 different mutations. Thus we can conclude that combining MLPA with DHPLC in point-mutations negative MO families, the detection of mutations in EXT genes can significantly improve the identification of both point-mutations and mid-size rearrangements. More important, we were able to characterize all those patients who were negative at the first PCR-based method screening.
authors.affiliation: Laboratory of Molecular Medicine and Biotechnology, University Campus Bio-Medico School of Medicine and Institute of Neurobiology and Molecular Medicine, CNR, Rome, Italy. e.signori@unicampus.it
authors.forename: Emanuela
authors.initials: E
authors.lastname: Signori
authors.forename: Emanuela
authors.initials: E
authors.lastname: Massi
authors.forename: Maria Giovanna
authors.initials: MG
authors.lastname: Matera
authors.forename: Monica
authors.initials: M
authors.lastname: Poscente
authors.forename: Carolina
authors.initials: C
authors.lastname: Gravina
authors.forename: Gianluca
authors.initials: G
authors.lastname: Falcone
authors.forename: Michele Attilio
authors.initials: MA
authors.lastname: Rosa
authors.forename: Monica
authors.initials: M
authors.lastname: Rinaldi
authors.forename: Wim
authors.initials: W
authors.lastname: Wuyts
authors.forename: Davide
authors.initials: D
authors.lastname: Seripa
authors.forename: Bruno
authors.initials: B
authors.lastname: Dallapiccola
authors.forename: Vito Michele
authors.initials: VM
authors.lastname: Fazio
mesh_terms: D019943:Amino Acid Substitution; D001859:Bone Neoplasms / Q000235:genetics*; D015331:Cohort Studies; D004252:DNA Mutational Analysis; D004273:DNA, Neoplasm / Q000235:genetics; D014644:Genetic Variation; D006801:Humans; D007558:Italy; D009154:Mutation; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics*; D015831:Osteochondroma / Q000235:genetics*; D017384:Sequence Deletion; D000099065:Exostosin 2; D000099058:Exostosin 1
chemical_list: D004273:DNA, Neoplasm; D017351:N-Acetylglucosaminyltransferases; D000099058:Exostosin 1; D000099065:Exostosin 2