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Identification of novel L1CAM mutations using fluorescence‐assisted mismatch analysis
Author(s) -
SaugierVeber Pascale,
Martin Cosette,
Le Meur Nathalie,
Lyonnet Stanislas,
Munnich Arnold,
David Albert,
Hénocq Alain,
Héron Delphine,
Jonveaux Philippe,
Odent Sylvie,
Manouvrier Sylvie,
Moncla Anne,
Morichon Nicole,
Philip Nicole,
Satge Daniel,
Tosi Mario,
Frébourg T
Publication year - 1998
Publication title -
human mutation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 162
eISSN - 1098-1004
pISSN - 1059-7794
DOI - 10.1002/(sici)1098-1004(1998)12:4<259::aid-humu7>3.0.co;2-a
Subject(s) - xq28 , biology , exon , genetics , gene , dna sequencer , coding region , genomic dna , allelic heterogeneity , mutation , microbiology and biotechnology , locus (genetics) , dna sequencing
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, performed with the two dye‐labeled universal primers, allowed us to generate 1‐kb‐labeled fragments, which were then submitted to the chemical clivage 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. Hum Mutat 12:259–266, 1998. © 1998 Wiley‐Liss, Inc.