title: Use of multilocus methylation-specific single nucleotide primer extension (MS-SNuPE) technology in diagnostic testing for human imprinted loci.
abstract: A number of diseases have been found to be linked to aberrant methylation of specific genes. However, most of the routine diagnostic techniques to detect epigenetic disturbances are restricted to single loci. Additionally, a precise quantification of the methylation status is often hampered. A considerable fraction of patients with Silver-Russell syndrome, Beckwith-Wiedemann syndrome and transient neonatal diabetes mellitus exhibit loss of methylation at further imprinted loci in addition to the disease specific ones (multilocus methylation defects, MLMD). As the currently available tests are mainly focused on single imprinted loci on different chromosomes and thereby make the detection of multilocus methylation defects time-consuming and expensive, we established methylation-specific single nucleotide primer extension (MS-SNuPE) assays for a simultaneous quantification of methylation at multiple methylated loci. We chose loci generally affected in patients with MLMD. The method was validated by screening 66 individuals with known (epi)genetic disturbances. In comparison to other methylation-specific techniques, multilocus methylation-specific single nucleotide primer extension allows the quantitative analysis of numerous CpG islands of different loci in one assay and is, therefore, suitable for the simultaneous diagnostic testing for different congenital imprinting disorders in parallel, as well as for MLMD.
authors.affiliation: Institute of Human Genetics, RWTH Aachen, Aachen, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Begemann
authors.forename: Isabelle
authors.initials: I
authors.lastname: Leisten
authors.forename: Lukas
authors.initials: L
authors.lastname: Soellner
authors.forename: Klaus
authors.initials: K
authors.lastname: Zerres
authors.forename: Thomas
authors.initials: T
authors.lastname: Eggermann
authors.forename: Sabrina
authors.initials: S
authors.lastname: Spengler
mesh_terms: D001506:Beckwith-Wiedemann Syndrome / Q000175:diagnosis / Q000235:genetics; D016022:Case-Control Studies; D002877:Chromosomes, Human / Q000235:genetics; D018899:CpG Islands; D019175:DNA Methylation; D017931:DNA Primers / Q000235:genetics; D044127:Epigenesis, Genetic; D030342:Genetic Diseases, Inborn / Q000175:diagnosis* / Q000235:genetics; D056426:Genetic Loci; D015894:Genome, Human; D018392:Genomic Imprinting; D006801:Humans; D058885:Multilocus Sequence Typing / Q000379:methods*; D009154:Mutation; D009711:Nucleotides / Q000235:genetics / Q000378:metabolism*; D015203:Reproducibility of Results; D056730:Silver-Russell Syndrome / Q000175:diagnosis / Q000235:genetics; D024182:Uniparental Disomy / Q000175:diagnosis / Q000235:genetics
chemical_list: D017931:DNA Primers; D009711:Nucleotides