Methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) robustly detects and distinguishes 11p15 abnormalities associated with overgrowth and growth retardation

• Published in: Journal of medical genetics Vol. 45; no. 2; pp. 106 - 113
• Main Authors: Scott, R H, Douglas, J, Baskcomb, L, Nygren, A O, Birch, J M, Cole, T R, Cormier-Daire, V, Eastwood, D M, Garcia-Minaur, S, Lupunzina, P, Tatton-Brown, K, Bliek, J, Maher, E R, Rahman, N
• Format: Journal Article
• Language: English
• Published: London BMJ Publishing Group Ltd 01.02.2008; BMJ; BMJ Publishing Group LTD
• Subjects: Beckwith-Wiedemann Syndrome - genetics; Biological and medical sciences; Chromosome Aberrations; Chromosomes, Human, Pair 11 - genetics; DNA Methylation; Epigenesis, Genetic; Epigenetics; Female; Fundamental and applied biological sciences. Psychology; Gene Dosage; General aspects. Genetic counseling; Genes; Genetic testing; Genetics of eukaryotes. Biological and molecular evolution; Genomic Imprinting; Growth Disorders - genetics; Humans; Male; Medical genetics; Medical sciences; Microsatellite Repeats; Molecular and cellular biology; Molecular Probe Techniques; Mutation; Nucleic Acid Amplification Techniques; Human; Gene amplification; Overgrowth syndrome; Association; Ligature; Genetics; Anomaly; Molecular probe; Methylation; Growth retardation
• ISSN: 0022-2593; 1468-6244; 1468-6244
• DOI: 10.1136/jmg.2007.053207

Background:A variety of abnormalities have been demonstrated at chromosome 11p15 in individuals with overgrowth and growth retardation. The identification of these abnormalities is clinically important but often technically difficult. Methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) is a simple but effective technique able to identify and differentiate methylation and copy number abnormalities, and thus is potentially well suited to the analysis of 11p15.Aims:To customise and test an MS-MLPA assay capable of detecting and distinguishing the full spectrum of known 11p15 epigenetic and copy number abnormalities associated with overgrowth and growth retardation and to assess its effectiveness as a first line investigation of these abnormalities.Methods:Five synthetic probe pairs were designed to extend the range of abnormalities detectable with a commercially available MS-MLPA assay. To define the normal values, 75 normal control samples were analysed using the customised assay. The assay was then used to analyse a “test set” of 24 normal and 27 abnormal samples, with data analysed by two independent blinded observers. The status of all abnormal samples was confirmed by a second technique.Results:The MS-MLPA assay gave reproducible, accurate methylation and copy number results in the 126 samples assayed. The blinded observers correctly identified and classified all 51 samples in the test set.Conclusions:MS-MLPA robustly and sensitively detects and distinguishes epigenetic and copy number abnormalities at 11p15 and is an effective first line investigation of 11p15 in individuals with overgrowth or growth retardation.