DHPLC analysis of the MECP2 gene in Italian Rett patients

Citation
P. Nicolao et al., DHPLC analysis of the MECP2 gene in Italian Rett patients, HUM MUTAT, 18(2), 2001, pp. 132-140
Citations number
37
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Molecular Biology & Genetics
Journal title
HUMAN MUTATION
ISSN journal
1059-7794 → ACNP
Volume
18
Issue
2
Year of publication
2001
Pages
132 - 140
Database
ISI
SICI code
1059-7794(2001)18:2<132:DAOTMG>2.0.ZU;2-F
Abstract
Rett Syndrome (RTT) is an X-linked dominant neurodevelopmental disorder, wh ich almost exclusively affects girls, with an estimated prevalence of one i n 10,000-15,000 female births. Mutations in the methyl CpG binding protein 2 gene (MECP2) have been identified in roughly 75% of classical Rett girls. The vast majority of Rett cases (99%) are sporadic in origin, and are due to de novo mutations. We collected DNA samples from 50 Italian classical Re tt girls, and screened the MECP2 coding region for mutations by denaturing high-performance liquid chromatography (DHPLC) and subsequent direct sequen cing. DHPLC is a recently developed method for mutation screening which ide ntifies heteroduplexes formed in DNA samples containing mismatches between wild type and mutant DNA strands, combining high sensitivity, reduced cost per run, and high throughput. In our series, 19 different de novo MECP2 mut ations, eight of which were previously unreported, were found in 35 out of 50 Rett girls (70%). Seven recurrent mutations were characterized in a tota l of 22 unrelated cases. Initial DHPLC screening allowed the identification of 17 out of 19 different mutations (90%); after optimal conditions were e stablished, this figure increased to 100%, with all recurrent MECP2 mutatio ns generating a characteristic chromatographic profile. Detailed clinical d ata were available for 27 out of 35 mutation carrying Rett girls. Milder di sease was detectable in patients carrying nonsense mutation as compared to patients carrying missense mutations, although this difference was not stat istically significant (P = 0.077). Hum Mutat 18.132-140, 2001. (C) 2001 Wil ey-Liss, Inc.