{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,20]],"date-time":"2026-07-20T10:50:47Z","timestamp":1784544647043,"version":"3.55.0"},"reference-count":25,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T00:00:00Z","timestamp":1777420800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Pediatric-onset neuromuscular diseases (NMDs) represent a clinically and genetically heterogeneous group of rare disorders, often posing significant diagnostic challenges due to overlapping phenotypes. Next-generation sequencing (NGS), particularly whole-exome sequencing (WES), has transformed the diagnostic landscape; however, its clinical utility varies across phenotypic subgroups. We conducted a combined retrospective\u2013prospective cohort study that included 100 pediatric patients with suspected neuromuscular disorders evaluated at a tertiary referral center between 2015 and 2025. Patients were stratified into seven clinically defined diagnostic categories prior to genetic testing. NGS-based diagnostics (primarily WES) were performed following initial clinical and targeted evaluations. Diagnostic yield and predictors of a positive genetic result were analyzed using univariate and multivariable statistical approaches. A molecular diagnosis was established in 71% of patients, including 64% with pathogenic\/likely pathogenic variants and 7% with clinically consistent variants of uncertain significance. Diagnostic yield varied significantly across disease categories (p &lt; 0.001), reaching near-complete rates in well-defined phenotypes such as congenital myasthenic syndromes, neuropathies, and metabolic myopathies, while markedly lower yield was observed in unclassified cases (38.2%). Multivariable logistic regression identified disease group as the only independent predictor of diagnostic success (B = \u22120.436, p = 0.001). Frequently implicated genes included DMD, RYR1, and LAMA2, reflecting a predominance of structural and excitation\u2013contraction coupling defects. NGS demonstrates high diagnostic utility in pediatric neuromuscular disorders, particularly when applied in a phenotype-driven framework. Diagnostic performance is strongly influenced by the degree of clinical definition prior to testing, highlighting the continued importance of expert phenotyping in the genomic era.<\/jats:p>","DOI":"10.3390\/ijms27093949","type":"journal-article","created":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T09:46:11Z","timestamp":1777455971000},"page":"3949","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Neuromuscular Disorders in Children Through the Lens of Next-Generation Sequencing: A Study of Diagnostic Yield"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8979-0445","authenticated-orcid":false,"given":"Slavica","family":"Ostoji\u0107","sequence":"first","affiliation":[{"name":"Department of Neurology, Mother and Child Health Care Institute of Serbia \u201cDr Vukan \u010cupi\u0107\u201d, 11070 Belgrade, Serbia"},{"name":"Faculty of Medicine, University of Belgrade, 11070 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0755-2153","authenticated-orcid":false,"given":"Gordana","family":"Kova\u010devi\u0107","sequence":"additional","affiliation":[{"name":"Department of Neurology, Mother and Child Health Care Institute of Serbia \u201cDr Vukan \u010cupi\u0107\u201d, 11070 Belgrade, Serbia"},{"name":"Faculty of Medicine, University of Belgrade, 11070 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1186-208X","authenticated-orcid":false,"given":"Nikola","family":"Ili\u0107","sequence":"additional","affiliation":[{"name":"Division of Clinical Genetics, Mother and Child Health Care Institute of Serbia \u201cDr Vukan \u010cupi\u0107\u201d, 11070 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3143-3098","authenticated-orcid":false,"given":"Nina","family":"Mari\u0107","sequence":"additional","affiliation":[{"name":"Clinic for Children Diseases, University Clinical Center of the Republic of Srpska, 78000 Banja Luka, Bosnia and Herzegovina"},{"name":"Medical Faculty, University of Banja Luka, 78000 Banja Luka, Bosnia and Herzegovina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4086-6197","authenticated-orcid":false,"given":"Marina","family":"An\u0111elkovi\u0107","sequence":"additional","affiliation":[{"name":"Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, 11070 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tanja","family":"Lali\u0107","sequence":"additional","affiliation":[{"name":"Laboratory of Medical Genetic, Mother and Child Health Care Institute of Serbia \u201cDr Vukan \u010cupi\u0107\u201d, 11070 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marijana","family":"Mi\u0161kovi\u0107","sequence":"additional","affiliation":[{"name":"Laboratory of Medical Genetic, Mother and Child Health Care Institute of Serbia \u201cDr Vukan \u010cupi\u0107\u201d, 11070 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3077-6091","authenticated-orcid":false,"given":"Kristel Klaassen","family":"Ljubi\u010di\u0107","sequence":"additional","affiliation":[{"name":"Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, 11070 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9102-5251","authenticated-orcid":false,"given":"Irena","family":"Marjanovi\u0107","sequence":"additional","affiliation":[{"name":"Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, 11070 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aleksandra","family":"Paripovi\u0107","sequence":"additional","affiliation":[{"name":"Department of Nephrology, Mother and Child Health Care Institute of Serbia \u201cDr Vukan \u010cupi\u0107\u201d, 11070 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1420-6072","authenticated-orcid":false,"given":"Mihail","family":"Ba\u0161a","sequence":"additional","affiliation":[{"name":"Department of Pulmology, Mother and Child Health Care Institute of Serbia \u201cDr Vukan \u010cupi\u0107\u201d, 11070 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vladislav","family":"Vukomanovi\u0107","sequence":"additional","affiliation":[{"name":"Faculty of Medicine, University of Belgrade, 11070 Belgrade, Serbia"},{"name":"Department of Cardiology, Mother and Child Health Care Institute of Serbia \u201cDr Vukan \u010cupi\u0107\u201d, 11070 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jovana","family":"Krsti\u0107","sequence":"additional","affiliation":[{"name":"Pediatric Clinic, Mother and Child Health Care Institute of Serbia \u201cDr Vukan \u010cupi\u0107\u201d, 11070 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Milica","family":"Adamovi\u0107","sequence":"additional","affiliation":[{"name":"Pediatric Clinic, Mother and Child Health Care Institute of Serbia \u201cDr Vukan \u010cupi\u0107\u201d, 11070 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aleksandar","family":"Sovti\u0107","sequence":"additional","affiliation":[{"name":"Faculty of Medicine, University of Belgrade, 11070 Belgrade, Serbia"},{"name":"Department of Pulmology, Mother and Child Health Care Institute of Serbia \u201cDr Vukan \u010cupi\u0107\u201d, 11070 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adrijan","family":"Sarajlija","sequence":"additional","affiliation":[{"name":"Faculty of Medicine, University of Belgrade, 11070 Belgrade, Serbia"},{"name":"Division of Clinical Genetics, Mother and Child Health Care Institute of Serbia \u201cDr Vukan \u010cupi\u0107\u201d, 11070 Belgrade, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.pediatrneurol.2023.08.034","article-title":"Pediatric neuromuscular diseases","volume":"149","author":"Rathore","year":"2023","journal-title":"Pediatr. 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