Multiplex Ligation-Dependent Probe Amplification (MLPA) Detects Large Deletions in the MECP2 Gene of Swedish Rett Syndrome Patients
- 1 December 2003
- journal article
- Published by Mary Ann Liebert Inc in Genetic Testing
- Vol. 7 (4) , 329-332
- https://doi.org/10.1089/109065703322783707
Abstract
Mutations in the methyl-CpG-binding protein-2 (MECP2) gene on Xq28 have been found to be a cause of Rett syndrome (RS). In a previous mutation screening, we found MECP2 mutations in 81% of Swedish classical Rett women. In this study, we have analyzed 22 patients for MECP2 deletions using multiplex-ligation-dependent probe amplification (MLPA). Clinically, 11 of the patients who were classical Rett women, 3 were forme fruste, 1 was congenital RS, and 7 were Rett variants. As inclusion criteria, we used DNA from patients in whom previous sequencing results showed no mutations in coding portions of the MECP2 gene. MLPA is a method based on multiplex PCR. In one PCR, as many as 40 probes are amplified with the same primers. The specificity of the amplification products is determined by the site-specific hybridization of each probe construct, prior to amplification. Each PCR product has a unique length, which makes it possible to identify it by size separation. In 3 of 11 (27%) classical Rett women, we detected large deletions in MECP2 using MLPA. All these patients had deletions covering two exons; in 2 cases the deletion involved exons 3 and 4 and, in one case, exons 1 and 2 were missing. In the forme fruste, congenital and Rett-variant patients, we found no large deletions. We have found that MLPA is useful when it comes to finding large deletions compromising whole exons in MECP2. Used as a complementary method to DNA sequencing, it revealed new MECP2 mutations in classical RS patients.Keywords
This publication has 13 references indexed in Scilit:
- Relative quantification of 40 nucleic acid sequences by multiplex ligation-dependent probe amplificationNucleic Acids Research, 2002
- Deletion screening by fluorescence in situ hybridization in Rett syndrome patients.Annales de Genetique, 2001
- MeCP2 mutations in children with and without the phenotype of Rett syndromeNeurology, 2001
- MECP2 mutation screening in Swedish classical Rett syndrome females.European Child & Adolescent Psychiatry, 2001
- MECP2 mutations in Danish patients with Rett syndrome: High frequency of mutations but no consistent correlations with clinical severity or with the X chromosome inactivation patternEuropean Journal of Human Genetics, 2001
- Rett syndrome: analysis of MECP2 and clinical characterization of 31 patients.Human Molecular Genetics, 2000
- Long-read sequence analysis of the MECP2 gene in Rett syndrome patients: correlation of disease severity with mutation type and locationHuman Molecular Genetics, 2000
- Rett syndrome: Methyl-CpG-binding protein 2 mutations and phenotype-genotype correlationsAmerican Journal of Medical Genetics, 2000
- Rett syndrome is caused by mutations in X-linked MECP2, encoding methyl-CpG-binding protein 2Nature Genetics, 1999
- Dissection of the methyl-CpG binding domain from the chromosomal protein MeCP2Nucleic Acids Research, 1993