{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:44:52Z","timestamp":1760060692348,"version":"build-2065373602"},"reference-count":31,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2025,9,11]],"date-time":"2025-09-11T00:00:00Z","timestamp":1757548800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"The Research Committee of the University of Thessaly to the Medical School Laboratory of Immunology and Histocompatibility"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Medicina"],"abstract":"<jats:p>Background and Objectives: Severe combined immunodeficiency (SCID) represents a group of rare and potentially fatal monogenic disorders arising from pathogenic variants in a broad spectrum of genes. Diagnostic delays beyond the first few months of life have been associated with poor overall survival and hematopoietic stem cell transplantation (HSCT) outcomes. Therefore, the aim of our study was to apply an NGS assay enabling the rapid and reliable diagnosis of SCID. Materials and Methods: We developed a targeted NGS panel of 30 genes implicated in the pathogenesis of most SCID cases and we applied it to three Greek infants with suspected SCID. Results: Each patient displayed a distinct immunophenotype\u2014T\u2212B\u2212NK\u2212, T\u2212B\u2212NK+ and T\u2212B+NK\u2212, respectively\u2014and was found to harbor pathogenic or likely pathogenic variants in the analyzed SCID-related genes. In particular, patient 1 carried two heterozygous ADA variants (c.58G&gt;A, p.Gly20Arg and c.956_960del, p.Glu319Glyfs); patient 2 harbored two discrete pathogenic variants in the DCLRE1C gene (a large deletion of exons 1\u20133 and the nonsense mutation c.241C&gt;T, p.Arg81*), causing Artemis deficiency; and patient 3 carried a hemizygous IL2RG missense variant (c.437T&gt;C, p.Leu146Pro), associated with X-linked SCID. All variants were confirmed by Sanger sequencing. Conclusions: Our method successfully identified the underlying genetic defects in all patients, thereby establishing a molecular diagnosis of SCID. These findings highlight the potential of targeted NGS assays for achieving rapid and accurate molecular diagnosis of SCID, which is crucial for the timely treatment of life-threatening conditions in affected children.<\/jats:p>","DOI":"10.3390\/medicina61091644","type":"journal-article","created":{"date-parts":[[2025,9,11]],"date-time":"2025-09-11T07:51:46Z","timestamp":1757577106000},"page":"1644","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Targeted Next-Generation Sequencing in the Molecular Diagnosis of Severe Combined Immunodeficiency"],"prefix":"10.3390","volume":"61","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1670-6895","authenticated-orcid":false,"given":"Evangelos","family":"Bakaros","sequence":"first","affiliation":[{"name":"Department of Immunology and Histocompatibility, Faculty of Medicine, University of Thessaly, 41500 Larissa, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8134-9228","authenticated-orcid":false,"given":"Styliani","family":"Sarrou","sequence":"additional","affiliation":[{"name":"Department of Immunology and Histocompatibility, Faculty of Medicine, University of Thessaly, 41500 Larissa, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3609-1824","authenticated-orcid":false,"given":"Antonios","family":"Gkantaras","sequence":"additional","affiliation":[{"name":"Pediatric Immunology and Rheumatology Referral Centre, First Department of Pediatrics, Aristotle University, \u201cHippokration\u201d General Hospital, 54642 Thessaloniki, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexia","family":"Matziri","sequence":"additional","affiliation":[{"name":"Laboratory of Hygiene and Epidemiology, Faculty of Medicine, University of Thessaly, 41222 Larissa, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Achilleas P.","family":"Galanopoulos","sequence":"additional","affiliation":[{"name":"Department of Immunology and Histocompatibility, Faculty of Medicine, University of Thessaly, 41500 Larissa, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2528-6426","authenticated-orcid":false,"given":"Konstantina","family":"Charisi","sequence":"additional","affiliation":[{"name":"Pediatric Immunology and Rheumatology Referral Centre, First Department of Pediatrics, Aristotle University, \u201cHippokration\u201d General Hospital, 54642 Thessaloniki, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Athanasios","family":"Bangeas","sequence":"additional","affiliation":[{"name":"Pediatric Immunology and Rheumatology Referral Centre, First Department of Pediatrics, Aristotle University, \u201cHippokration\u201d General Hospital, 54642 Thessaloniki, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5083-6824","authenticated-orcid":false,"given":"Anna","family":"Taparkou","sequence":"additional","affiliation":[{"name":"Pediatric Immunology and Rheumatology Referral Centre, First Department of Pediatrics, Aristotle University, \u201cHippokration\u201d General Hospital, 54642 Thessaloniki, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eleni","family":"Papadimitriou","sequence":"additional","affiliation":[{"name":"Pediatric Immunology and Rheumatology Referral Centre, First Department of Pediatrics, Aristotle University, \u201cHippokration\u201d General Hospital, 54642 Thessaloniki, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Varvara A.","family":"Mouchtouri","sequence":"additional","affiliation":[{"name":"Laboratory of Hygiene and Epidemiology, Faculty of Medicine, University of Thessaly, 41222 Larissa, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fani","family":"Kalala","sequence":"additional","affiliation":[{"name":"Department of Immunology and Histocompatibility, Faculty of Medicine, University of Thessaly, 41500 Larissa, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4769-8376","authenticated-orcid":false,"given":"Christos","family":"Hadjichristodoulou","sequence":"additional","affiliation":[{"name":"Laboratory of Hygiene and Epidemiology, Faculty of Medicine, University of Thessaly, 41222 Larissa, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1287-7734","authenticated-orcid":false,"given":"Matthaios","family":"Speletas","sequence":"additional","affiliation":[{"name":"Department of Immunology and Histocompatibility, Faculty of Medicine, University of Thessaly, 41500 Larissa, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5132-8255","authenticated-orcid":false,"given":"Evangelia","family":"Farmaki","sequence":"additional","affiliation":[{"name":"Pediatric Immunology and Rheumatology Referral Centre, First Department of Pediatrics, Aristotle University, \u201cHippokration\u201d General Hospital, 54642 Thessaloniki, Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e20250003","DOI":"10.70962\/jhi.20250003","article-title":"Human inborn errors of immunity: 2024 update on the classification from the International Union of Immunological Societies Expert Committee","volume":"1","author":"Poli","year":"2025","journal-title":"J. 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