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. 2025:2866:45-57.
doi: 10.1007/978-1-0716-4192-7_3.

Targeted Next-Generation Sequencing in Rare Diseases

Aleša Kristan  1 , Nataša Debeljak  2
Affiliations

Targeted Next-Generation Sequencing in Rare Diseases

Aleša Kristan et al. Methods Mol Biol. 2025.

Abstract

Targeted next-generation sequencing (NGS) in rare disease focuses on genetic analysis of specific regions in genome that are linked to a rare disease. In addition to library preparation, sequencing, and data analysis, targeted NGS includes an additional step of target enrichment of selected genes and regions. It allows for more sensitive and profound sequencing, as it is a fast and cost-effective approach with less data burden and is therefore often a method of choice for identifying rare variants in known genes, especially in diagnostics of rare diseases. Several in silico tools address the pathogenicity predictions of rare variants of unknown significance (VUS) and can therefore facilitate clinical interpretation.

Keywords: Data analysis; Disease-specific DNA probes; Hybridization capture; Library preparation; Pathogenicity prediction; Sequencing; Targeted enrichment.

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