title: Methylation-specific MLPA (MS-MLPA): simultaneous detection of CpG methylation and copy number changes of up to 40 sequences.
abstract: Copy number changes and CpG methylation of various genes are hallmarks of tumor development but are not yet widely used in diagnostic settings. The recently developed multiplex ligation-dependent probe amplification (MLPA) method has increased the possibilities for multiplex detection of gene copy number aberrations in a routine laboratory. Here we describe a novel robust method: the methylation-specific MLPA (MS-MLPA) that can detect changes in both CpG methylation as well as copy number of up to 40 chromosomal sequences in a simple reaction. In MS-MLPA, the ligation of MLPA probe oligonucleotides is combined with digestion of the genomic DNA-probe hybrid complexes with methylation-sensitive endonucleases. Digestion of the genomic DNA-probe complex, rather than double-stranded genomic DNA, allowed the use of DNA derived from the formalin treated paraffin-embedded tissue samples, enabling retrospective studies. To validate this novel method, we used MS-MLPA to detect aberrant methylation in DNA samples of patients with Prader-Willy syndrome, Angelman syndrome or acute myeloid leukemia.
authors.affiliation: MRC-Holland bv Amsterdam, The Netherlands.
authors.forename: Anders O H
authors.initials: AO
authors.lastname: Nygren
authors.forename: Najim
authors.initials: N
authors.lastname: Ameziane
authors.forename: Helena M B
authors.initials: HM
authors.lastname: Duarte
authors.forename: Raymon N C P
authors.initials: RN
authors.lastname: Vijzelaar
authors.forename: Quinten
authors.initials: Q
authors.lastname: Waisfisz
authors.forename: Corine J
authors.initials: CJ
authors.lastname: Hess
authors.forename: Jan P
authors.initials: JP
authors.lastname: Schouten
authors.forename: Abdellatif
authors.initials: A
authors.lastname: Errami
mesh_terms: D000208:Acute Disease; D017204:Angelman Syndrome / Q000235:genetics; D045744:Cell Line, Tumor; D018899:CpG Islands; D019175:DNA Methylation; D018628:Gene Dosage; D006801:Humans; D007951:Leukemia, Myeloid / Q000175:diagnosis / Q000235:genetics; D025202:Molecular Diagnostic Techniques; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction / Q000379:methods*; D011218:Prader-Willi Syndrome / Q000235:genetics; D017422:Sequence Analysis, DNA; D013447:Sulfites / Q000737:chemistry
chemical_list: D013447:Sulfites