title: Genetic testing for monogenic diabetes using targeted next-generation sequencing in patients with maturity-onset diabetes of the young.
abstract: BACKGROUND
  Molecular diagnosis of monogenic diabetes mellitus is important for individualized patient care. Next-generation sequencing (NGS) enables a simultaneous analysis of multiple genes in a single test.
  
  
  OBJECTIVE
  We aimed to assess the feasibility of using NGS for detecting mutations in a set of known monogenic diabetes gene mutations in a cohort of Polish patients with maturity-onset diabetes of the young (MODY) with earlier negative Sanger sequencing results for HNF1A-MODY or GCK-MODY.
  
  
  METHODS
  We selected a panel of 28 chromosomal genes in which mutations have been reported to cause monogenic diabetes. The MiSeq platform was used for NGS. An exon-capture assay was designed to include coding regions and splice sites. A total of 54 patients with existing negative Sanger sequencing screening results for HNF1A or GCK gene mutations were selected for the study.
  
  
  RESULTS
  NGS results were generated for all 54 patients and 9 positive controls with previously identified HNF1A or GCK gene mutation. All selected positive controls were confirmed by NGS. Among 28 genes, mutations were detected in 16. The type of the analyzed genetic changes was described in the NGS study as high (n = 3) or moderate (n = 76). Among the detected mutations, there were 4 known GCK gene mutations that had been previously missed in Sanger sequencing. So far, Sanger sequencing allowed us to confirm 21 gene mutations detected by NGS, and segregation with diabetes in 14 pedigrees.
  
  
  CONCLUSIONS
  Our pilot study using NGS for monogenic diabetes screening in the MODY cohort confirmed that it improves the detection of diabetes-related sequence differences. The screening with NGS should also include diabetic patients for whom Sanger-based screening for particular subtypes of MODY provided negative results.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Szopa
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Ludwig-Gałęzowska
authors.forename: Piotr
authors.initials: P
authors.lastname: Radkowski
authors.forename: Jan
authors.initials: J
authors.lastname: Skupień
authors.forename: Barbara
authors.initials: B
authors.lastname: Zapała
authors.forename: Teresa
authors.initials: T
authors.lastname: Płatek
authors.forename: Tomasz
authors.initials: T
authors.lastname: Klupa
authors.forename: Beata
authors.initials: B
authors.lastname: Kieć-Wilk
authors.forename: Maciej
authors.initials: M
authors.lastname: Borowiec
authors.forename: Wojciech
authors.initials: W
authors.lastname: Młynarski
authors.forename: Paweł
authors.initials: P
authors.lastname: Wołkow
authors.forename: Maciej T
authors.initials: MT
authors.lastname: Małecki
mesh_terms: D000293:Adolescent; D000328:Adult; D002648:Child; D004252:DNA Mutational Analysis; D003924:Diabetes Mellitus, Type 2 / Q000175:diagnosis* / Q000235:genetics*; D005260:Female; D005820:Genetic Testing / Q000379:methods; D055106:Genome-Wide Association Study; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D008297:Male; D009154:Mutation; D010375:Pedigree; D010865:Pilot Projects; D017422:Sequence Analysis, DNA; D055815:Young Adult