title: Improved molecular diagnosis of patients with neonatal diabetes using a combined next-generation sequencing and MS-MLPA approach.
abstract: BACKGROUND
  We evaluated a methylation-specific multiplex-ligation-dependent probe amplification (MS-MLPA) assay for the molecular diagnosis of transient neonatal diabetes mellitus (TNDM) caused by 6q24 abnormalities and assessed the clinical utility of using this assay in combination with next generation sequencing (NGS) analysis for diagnosing patients with neonatal diabetes (NDM).
  
  
  METHODS
  We performed MS-MLPA in 18 control samples and 42 retrospective NDM cases with normal bi-parental inheritance of chromosome 6. Next, we evaluated 22 prospective patients by combining NGS analysis of 11 NDM genes and the MS-MLPA assay.
  
  
  RESULTS
  6q24 aberrations were identified in all controls and in 19% of patients with normal bi-parental inheritance of chromosome 6. The MS-MLPA/NGS combined approach identified a genetic cause in ~64% of patients with NDM of unknown etiology.
  
  
  CONCLUSIONS
  MS-MLPA is a reliable method to identify all known 6q24 abnormalities and comprehensive testing of all causes reveals a causal mutation in ~64% of patients.
authors.forename: Gorka
authors.initials: G
authors.lastname: Alkorta-Aranburu
authors.forename: Madina
authors.initials: M
authors.lastname: Sukhanova
authors.forename: David
authors.initials: D
authors.lastname: Carmody
authors.forename: Trevor
authors.initials: T
authors.lastname: Hoffman
authors.forename: Latrice
authors.initials: L
authors.lastname: Wysinger
authors.forename: Jennifer
authors.initials: J
authors.lastname: Keller-Ramey
authors.forename: Zejuan
authors.initials: Z
authors.lastname: Li
authors.forename: Amy Knight
authors.initials: AK
authors.lastname: Johnson
authors.forename: Frances
authors.initials: F
authors.lastname: Kobiernicki
authors.forename: Shaun
authors.initials: S
authors.lastname: Botes
authors.forename: Carrie
authors.initials: C
authors.lastname: Fitzpatrick
authors.forename: Soma
authors.initials: S
authors.lastname: Das
authors.forename: Daniela
authors.initials: D
authors.lastname: Del Gaudio
mesh_terms: D015415:Biomarkers / Q000378:metabolism*; D016022:Case-Control Studies; D019175:DNA Methylation; D003920:Diabetes Mellitus / Q000175:diagnosis* / Q000235:genetics; D005500:Follow-Up Studies; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D007231:Infant, Newborn; D007232:Infant, Newborn, Diseases / Q000175:diagnosis* / Q000235:genetics; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D016133:Polymerase Chain Reaction; D011379:Prognosis; D011446:Prospective Studies; D012189:Retrospective Studies
chemical_list: D015415:Biomarkers