title: Identification of novel L1CAM mutations using fluorescence-assisted mismatch analysis.
abstract: The L1CAM gene, which is located in Xq28 and codes for a neuronal cell adhesion molecule, is involved in three distinct conditions: HSAS (hydrocephalus-stenosis of the aqueduct of Sylvius), MASA (mental retardation, aphasia, shuffling gait, adductus thumbs), and SPG1 (spastic paraplegia). Molecular analysis of the L1CAM gene is labor-intensive because of the size of the coding region, which is fragmented in numerous exons, and because of the great allelic heterogeneity and distribution of the mutations. The FAMA (fluorescent assisted mismatch analysis) method combines the excellent sensitivity of the chemical cleavage method for scanning PCR fragments larger than 1 kb and the power of automated DNA sequencers. In order to optimize this method for L1CAM, we divided the gene into nine genomic fragments, each including three to four exons. These fragments were PCR-amplified using nine sets of primers containing additional rare universal sequences. A second-stage PCR, per formed with the two dye-labeled universal primers, allowed us to generate 1-kb-labeled fragments, which were then submitted to the chemical cleavage analysis. Among 12 French families with HSAS and/or MASA, we identified nine distinct L1CAM mutations, seven of which were novel, and an intronic variation. This study demonstrates that FAMA allows rapid and reliable detection of mutations in the L1CAM gene and thus represents one of the most appropriate methods to provide diagnosis for accurate genetic counseling in families with HSAS, MASA, or SPG1.
authors.affiliation: Laboratoire de Génétique Moléculaire, CHU de Rouen, France.
authors.forename: P
authors.initials: P
authors.lastname: Saugier-Veber
authors.forename: C
authors.initials: C
authors.lastname: Martin
authors.forename: N
authors.initials: N
authors.lastname: Le Meur
authors.forename: S
authors.initials: S
authors.lastname: Lyonnet
authors.forename: A
authors.initials: A
authors.lastname: Munnich
authors.forename: A
authors.initials: A
authors.lastname: David
authors.forename: A
authors.initials: A
authors.lastname: Hénocq
authors.forename: D
authors.initials: D
authors.lastname: Héron
authors.forename: P
authors.initials: P
authors.lastname: Jonveaux
authors.forename: S
authors.initials: S
authors.lastname: Odent
authors.forename: S
authors.initials: S
authors.lastname: Manouvrier
authors.forename: A
authors.initials: A
authors.lastname: Moncla
authors.forename: N
authors.initials: N
authors.lastname: Morichon
authors.forename: N
authors.initials: N
authors.lastname: Philip
authors.forename: D
authors.initials: D
authors.lastname: Satge
authors.forename: M
authors.initials: M
authors.lastname: Tosi
authors.forename: T
authors.initials: T
authors.lastname: Frébourg
mesh_terms: D001037:Aphasia / Q000235:genetics; D020137:Base Pair Mismatch; D004252:DNA Mutational Analysis / Q000379:methods*; D005260:Female; D005453:Fluorescence; D014644:Genetic Variation / Q000235:genetics; D006801:Humans; D006849:Hydrocephalus / Q000235:genetics*; D008607:Intellectual Disability / Q000235:genetics*; D007438:Introns / Q000235:genetics; D039841:Leukocyte L1 Antigen Complex; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics*; D009069:Movement Disorders / Q000235:genetics; D009154:Mutation / Q000235:genetics; D019006:Neural Cell Adhesion Molecules / Q000235:genetics*; D009692:Nucleic Acid Heteroduplexes / Q000032:analysis*; D010264:Paraplegia / Q000235:genetics; D010375:Pedigree; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D013577:Syndrome; D013933:Thumb / Q000002:abnormalities
chemical_list: D039841:Leukocyte L1 Antigen Complex; D008562:Membrane Glycoproteins; D019006:Neural Cell Adhesion Molecules; D009692:Nucleic Acid Heteroduplexes