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Genetic testing for monogenic diabetes using targeted next-generation sequencing in patients with maturity-onset diabetes of the young
DOI: 10.20452/pamw.3164
Published online: November 09, 2015
Introduction 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.
Objectives 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.
Patient s and 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.
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