DEVELOPMENT AND OPTIMIZATION OF A TARGETED NGS PANEL FOR MODY DIAGNOSIS
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Authors
A. Toleuzhanova
National Center for Biotechnology, 13/5, Kurgalzhynskoye road, Astana, Kazakhstan, 010000
A. Romanova
National Center for Biotechnology, 13/5, Kurgalzhynskoye road, Astana, Kazakhstan, 010000;
E. Zholdybayeva
National Center for Biotechnology, 13/5, Kurgalzhynskoye road, Astana, Kazakhstan, 010000
R. Kalendar
National Laboratory Astana, Qabanbay Batyr Ave 53, Astana, Kazakhstan, 020000
Abstract
This study presents the development and optimization of a targeted next-generation sequencing (NGS) panel for the diagnosis of maturity-onset diabetes of the young (MODY). MODY is frequently misdiagnosed as type 1 or type 2 diabetes, highlighting the need for accurate genetic testing approaches.
This study aimed to design a primer panel targeting MODY-associated genes, optimize multiplex PCR conditions, and evaluate the performance of the developed workflow.
A total of 124 primer pairs were designed using PCRpanel software and tested. Specific amplification of target regions (300-400 bp) was confirmed by agarose gel electrophoresis. Multiplex PCR conditions were optimized. DNA libraries were prepared and successfully sequenced using the Illumina MiSeq platform. The minimum sequencing coverage achieved was 30×, with most regions exceeding 100×.
The developed panel demonstrated high specificity and may serve as an effective diagnostic tool for MODY.
Keywords
MODY, targeted panel, NGS, multiplex PCR, genetic diagnostics, WES
Article Details
References
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