{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,11]],"date-time":"2026-02-11T16:41:51Z","timestamp":1770828111106,"version":"3.50.1"},"reference-count":36,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T00:00:00Z","timestamp":1753833600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"\u201cFonds de Recherche Clinique\u201d of the Cliniques universitaires Saint-Luc, (\u201cCr\u00e9dits de recherche\u2014starting grant\u201d)"},{"name":"Fondation Saint Luc"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJNS"],"abstract":"<jats:p>Pyridoxine-dependent epilepsy (PDE) represents a group of rare developmental and epileptic encephalopathies. The most common PDE is caused by biallelic pathogenic variants in ALDH7A1 (PDE-ALDH7A1; OMIM #266100), which encodes \u03b1-aminoadipate semialdehyde (\u03b1-AASA) dehydrogenase, a key enzyme in lysine catabolism. Affected individuals present with seizures unresponsive to conventional anticonvulsant medications but responsive to high-dose of pyridoxine (vitamin B6). Adjunctive lysine restriction and arginine supplementation have also shown potential in improving neurodevelopmental outcomes. Given the significant benefit of early intervention, PDE-ALDH7A1 is a strong candidate for newborn screening (NBS). However, traditional biomarkers are biochemically unstable at room temperature (\u03b1-AASA and piperideine-6-carboxylate) or lack sufficient specificity (pipecolate), limiting their utility for biomarker-based NBS. The recent identification of two novel and stable biomarkers, 2S,6S-\/2S,6R-oxopropylpiperidine-2-carboxylate (2-OPP) and 6-oxo-pipecolate (oxo-PIP), offers renewed potential for biochemical NBS. We evaluated the feasibility of incorporating 2-OPP, oxo-PIP, and pipecolate into routine butylated FIA-MS\/MS workflows used for biochemical NBS. A total of 9402 dried blood spots (DBS), including nine confirmed PDE-ALDH7A1 patients and 9393 anonymized controls were analyzed using a single multiplex assay. 2-OPP emerged as the most sensitive biomarker, identifying all PDE-ALDH7A1 patients with 100% sensitivity and a positive predictive value (PPV) of 18.4% using a threshold above the 99.5th percentile. Combining elevated 2-OPP (above the 99.5th percentile) with either pipecolate or oxo-PIP (above the 85.0th percentile) as secondary marker detected within the same multiplex FIA-MS\/MS assay further improved the PPVs to 60% and 45%, respectively, while maintaining compatibility with butanol-derivatized method. Notably, increasing the 2-OPP threshold above the 99.89th percentile, in combination with either pipecolate or oxo-PIP above the 85.0th percentile resulted in both 100% sensitivity and 100% PPV. This study supports the strong potential of 2-OPP-based neonatal screening for PDE-ALDH7A1 within existing NBS infrastructures. The ability to multiplex 2-OPP, pipecolate and oxo-PIP within a single assay offers a robust, practical, high-throughput and cost-effective approach. These results support the inclusion of PDE-ALDH7A1 in existing biochemical NBS panels. Further prospective studies in larger cohorts are needed to refine cutoffs and confirm clinical performance.<\/jats:p>","DOI":"10.3390\/ijns11030059","type":"journal-article","created":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T10:55:37Z","timestamp":1753872937000},"page":"59","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Advancing Neonatal Screening for Pyridoxine-Dependent Epilepsy-ALDH7A1 Through Combined Analysis of 2-OPP, 6-Oxo-Pipecolate and Pipecolate in a Butylated FIA-MS\/MS Workflow"],"prefix":"10.3390","volume":"11","author":[{"given":"Myl\u00e8ne","family":"Donge","sequence":"first","affiliation":[{"name":"Department of Pediatric Neurology, Kannerklinik Centre Hospitalier du Luxembourg, L-1210 Luxembourg, Luxembourg"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sandrine","family":"Marie","sequence":"additional","affiliation":[{"name":"Biochemical Genetics and Newborn Screening Laboratory, Department of Laboratory Medicine, Cliniques Universitaires Saint-Luc, UCLouvain, B-1200 Brussels, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Amandine","family":"Pochet","sequence":"additional","affiliation":[{"name":"Louvain Centre for Toxicology and Applied Pharmacology, Institut de Recherche Experimentale et Clinique, UCLouvain, B-1200 Brussels, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lionel","family":"Marcelis","sequence":"additional","affiliation":[{"name":"Laboratory of Paediatric Research and Newborn Screening, Universit\u00e9 Libre de Bruxelles, B-1020 Brussels, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Geraldine","family":"Luis","sequence":"additional","affiliation":[{"name":"Biochemical Genetics and Newborn Screening Laboratory, CHU Liege, B-4000 Liege, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4470-9524","authenticated-orcid":false,"given":"Fran\u00e7ois","family":"Boemer","sequence":"additional","affiliation":[{"name":"Biochemical Genetics and Newborn Screening Laboratory, CHU Liege, B-4000 Liege, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2986-5599","authenticated-orcid":false,"given":"Cl\u00e9ment","family":"Prouteau","sequence":"additional","affiliation":[{"name":"Department of Medical Genetics, Angers University Hospital, F-49000 Angers, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Samir","family":"Mesli","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Centre Hospitalier Universitaire de Bordeaux, F-33404 Bordeaux, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3024-6939","authenticated-orcid":false,"given":"Matthias","family":"Cuykx","sequence":"additional","affiliation":[{"name":"Clinical Chemistry and Newborn Screening Center, UZ Antwerpen, B-2650 Antwerpen, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3738-7299","authenticated-orcid":false,"given":"Thao","family":"Nguyen-Khoa","sequence":"additional","affiliation":[{"name":"Laboratoire du Centre R\u00e9gional de D\u00e9pistage N\u00e9onatal de l\u2019Ile de France, H\u00f4pital Necker-Enfants Malades, AP-HP Centre Universit\u00e9 Paris Cit\u00e9, 161, rue de S\u00e8vres, F-75015 Paris, France"},{"name":"INSERM U1151, Institut Necker Enfants Malades, F-75015 Paris, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David","family":"Gu\u00e9net","sequence":"additional","affiliation":[{"name":"Laboratory of Biochemistry, Normandie Universit\u00e9, UNICAEN, CHU of Caen Normandie, F-14000 Caen, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aur\u00e9lie","family":"Empain","sequence":"additional","affiliation":[{"name":"Nutrition and Metabolic Clinic, Brussels University Hospital, Academic Children Hospital Queen Fabiola, Universit\u00e9 Libre de Bruxelles, B-1020 Brussels, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Magalie","family":"Barth","sequence":"additional","affiliation":[{"name":"Department of Medical Genetics, Angers University Hospital, F-49000 Angers, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Benjamin","family":"Dauriat","sequence":"additional","affiliation":[{"name":"Medical Genetics and Cytogenetics Department, Limoges Universitary Hospital, F-87000 Limoges, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"C\u00e9cile","family":"Laroche-Raynaud","sequence":"additional","affiliation":[{"name":"Centre de Comp\u00e9tence des Maladies H\u00e9r\u00e9ditaires du M\u00e9tabolisme, Centre Hospitalier Universitaire de Limoges, F-87000 Limoges, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Corinne","family":"De Laet","sequence":"additional","affiliation":[{"name":"Nutrition and Metabolic Clinic, Brussels University Hospital, Academic Children Hospital Queen Fabiola, Universit\u00e9 Libre de Bruxelles, B-1020 Brussels, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2077-0275","authenticated-orcid":false,"given":"Patrick","family":"Verloo","sequence":"additional","affiliation":[{"name":"Department of Pediatric Neurology, Center for Inherited Metabolic Disorders, University Hospital Ghent, B-9000 Ghent, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"An I.","family":"Jonckheere","sequence":"additional","affiliation":[{"name":"Centre for Metabolic Diseases, University Hospital Antwerp, University of Antwerp, B-2650 Antwerpen, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Manuel","family":"Schiff","sequence":"additional","affiliation":[{"name":"Reference Center for Inborn Errors of Metabolism, Necker University Hospital, AP-HP Centre Universit\u00e9 Paris Cit\u00e9, Fili\u00e8re G2M, F-75015 Paris, France"},{"name":"INSERM UMRS_1163, Institut Imagine, F-75015 Paris, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6936-1611","authenticated-orcid":false,"given":"Marie-C\u00e9cile","family":"Nassogne","sequence":"additional","affiliation":[{"name":"Department of Pediatric Neurology, Cliniques Universitaires Saint-Luc, UCLouvain, B-1200 Brussels, Belgium"},{"name":"Institut des 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