Journal Article

NGS Approaches in Clinical Diagnostics: From Workflow to Disease-Specific Applications

Treccarichi, Simone, Bruno, Francesca, Coniglio, Elvira, Brancato, Desiree, Saccone, Salvatore, Calì, Francesco, Vinci, Mirella, Federico, Concetta

2025

Overview

Next-Generation Sequencing (NGS) techniques have become a cornerstone of molecular diagnostics, enabling high-throughput, parallel analysis of multiple disease-associated genes. Their targeted design allows streamlined interpretation and optimised diagnostic yield, especially in disorders with known genetic heterogeneity. In this review, we provide a comprehensive overview of the clinical application of NGS techniques—targeted gene panels, whole exome sequencing (WES) and whole genome sequencing (WGS)—detailing the methodological workflow and the critical steps involved in their implementation. Particular emphasis is placed on the genes identified through NGS that are implicated in neurodevelopmental, neurodegenerative, psychiatric, neuromuscular, cardiovascular, and metabolic disorders. We also compare the advantages and limitations of panel-based diagnostics versus WES and WGS, and discuss future directions, including the integration of long-read sequencing technologies into multidisciplinary clinical practice. Finally, we consider how these advances may ultimately bridge biomedical research and clinical practise to improve the diagnosis and management of multifactorial diseases.

• Publisher: MDPI AG
• Subject: Accuracy / Connective tissue / Data analysis / Development and progression / Diagnostic equipment (Medical) / Disease / Diseases / DNA sequencing / Exome Sequencing - methods / Genes / Genomes / Genomics / Genotype & phenotype / High-Throughput Nucleotide Sequencing - methods / Humans / Italy / Medical diagnosis / Medical research / Medicine, Experimental / Metabolism / Molecular Diagnostic Techniques - methods / Nucleotide sequencing / Whole genome sequencing / Whole Genome Sequencing - methods / Workflow