Combined biochemical profiling and DNA sequencing in the expanded newborn screening for inherited metabolic diseases: the experience in an Italian reference center
Abstract Background Newborn screening (NBS) programs have significantly improved the health and outcomes of patients with inherited metabolic disorders (IMDs). Methods based on liquid chromatography/mass spectrometry (LC–MS/MS) analysis are viewed worldwide as the gold standard procedure for the exp...
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2025-01-01
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| author | Simona Fecarotta Lorenzo Vaccaro Alessandra Verde Marianna Alagia Alessandro Rossi Chiara Colantuono Maria Teresa Cacciapuoti Patrizia Annunziata Sara Riccardo Antonio Grimaldi Tonya Fusco Rosa De Santis Fernando Barretta Lucia Albano Daniela Crisci Fabiana Vallone Antonietta Tarallo Marcella Cesana Nicola Brunetti-Pierri Giulia Frisso Margherita Ruoppolo Davide Cacchiarelli Giancarlo Parenti |
| author_facet | Simona Fecarotta Lorenzo Vaccaro Alessandra Verde Marianna Alagia Alessandro Rossi Chiara Colantuono Maria Teresa Cacciapuoti Patrizia Annunziata Sara Riccardo Antonio Grimaldi Tonya Fusco Rosa De Santis Fernando Barretta Lucia Albano Daniela Crisci Fabiana Vallone Antonietta Tarallo Marcella Cesana Nicola Brunetti-Pierri Giulia Frisso Margherita Ruoppolo Davide Cacchiarelli Giancarlo Parenti |
| author_sort | Simona Fecarotta |
| collection | DOAJ |
| description | Abstract Background Newborn screening (NBS) programs have significantly improved the health and outcomes of patients with inherited metabolic disorders (IMDs). Methods based on liquid chromatography/mass spectrometry (LC–MS/MS) analysis are viewed worldwide as the gold standard procedure for the expanded NBS programs for these disorders. Advanced molecular technologies point to genomic sequencing as an alternative and feasible strategy for the screening of genetic diseases, including IMDs. However, each of the two approaches has potential limitations when used as a first-tier analysis. In this study, we tested a workflow-based parallel biochemical and sequencing analyses to determine whether this approach could improve the diagnostic outcome. Results For each patient identified by LC–MS/MS as positive, we performed both the biochemical confirmatory tests and next-generation sequencing (NGS) procedures from the same Dried Blood Spot (DBS). NGS analysis was based on applying Exome Sequencing libraries, limiting the analysis to 105 actionable genes involved in IMDs. This allows overtaking the actual limitations of NBS on DBS, enhancing our capacity to identify variants that can drive a genetic disease. Through this approach, we could reach 100% of cases solved, with 37.9% of cases (41/108) for which the combination of the biochemical and NGS analysis was indispensable for a correct diagnosis. In total, we could identify 17 affected, 34 false positives, 12 individuals referred to us for maternal conditions. In 45 newborns the molecular analysis showed heterozygosity for mutations in one or more of the genes analyzed, with results compatible with the biochemical profile indicative of NBS positivity. Conclusions In this study, we validated the performance of the proposed workflow. The advantage of this approach is limiting molecular analysis only to positive newborns and using a restricted panel of 105 genes relevant for the expanded NBS, with a 100% rate of diagnosis and potential reduction of the costs related to NBS procedures and reduced impact on patients and families. |
| format | Article |
| id | doaj-art-be2db783765c47a39f414a19fae526fc |
| institution | Kabale University |
| issn | 1750-1172 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | BMC |
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| series | Orphanet Journal of Rare Diseases |
| spelling | doaj-art-be2db783765c47a39f414a19fae526fc2025-01-26T12:52:16ZengBMCOrphanet Journal of Rare Diseases1750-11722025-01-0120111010.1186/s13023-025-03546-1Combined biochemical profiling and DNA sequencing in the expanded newborn screening for inherited metabolic diseases: the experience in an Italian reference centerSimona Fecarotta0Lorenzo Vaccaro1Alessandra Verde2Marianna Alagia3Alessandro Rossi4Chiara Colantuono5Maria Teresa Cacciapuoti6Patrizia Annunziata7Sara Riccardo8Antonio Grimaldi9Tonya Fusco10Rosa De Santis11Fernando Barretta12Lucia Albano13Daniela Crisci14Fabiana Vallone15Antonietta Tarallo16Marcella Cesana17Nicola Brunetti-Pierri18Giulia Frisso19Margherita Ruoppolo20Davide Cacchiarelli21Giancarlo Parenti22Department of Translational Medicine, University of Naples “Federico II”Department of Translational Medicine, University of Naples “Federico II”Department of Translational Medicine, University of Naples “Federico II”Department of Translational Medicine, University of Naples “Federico II”Department of Translational Medicine, University of Naples “Federico II”NEGEDIA SrlNEGEDIA SrlNEGEDIA SrlNEGEDIA SrlDepartment of Translational Medicine, University of Naples “Federico II”Department of Translational Medicine, University of Naples “Federico II”Department of Translational Medicine, University of Naples “Federico II”Department of Molecular Medicine and Medical Biotechnology, University of Naples “Federico II”CEINGE Advanced Biotechnologies Franco SalvatoreCEINGE Advanced Biotechnologies Franco SalvatoreCEINGE Advanced Biotechnologies Franco SalvatoreDepartment of Translational Medicine, University of Naples “Federico II”Telethon Institute of Genetics and Medicine (TIGEM), Armenise/Harvard Laboratory of Integrative GenomicsDepartment of Translational Medicine, University of Naples “Federico II”Department of Molecular Medicine and Medical Biotechnology, University of Naples “Federico II”Department of Molecular Medicine and Medical Biotechnology, University of Naples “Federico II”Department of Translational Medicine, University of Naples “Federico II”Department of Translational Medicine, University of Naples “Federico II”Abstract Background Newborn screening (NBS) programs have significantly improved the health and outcomes of patients with inherited metabolic disorders (IMDs). Methods based on liquid chromatography/mass spectrometry (LC–MS/MS) analysis are viewed worldwide as the gold standard procedure for the expanded NBS programs for these disorders. Advanced molecular technologies point to genomic sequencing as an alternative and feasible strategy for the screening of genetic diseases, including IMDs. However, each of the two approaches has potential limitations when used as a first-tier analysis. In this study, we tested a workflow-based parallel biochemical and sequencing analyses to determine whether this approach could improve the diagnostic outcome. Results For each patient identified by LC–MS/MS as positive, we performed both the biochemical confirmatory tests and next-generation sequencing (NGS) procedures from the same Dried Blood Spot (DBS). NGS analysis was based on applying Exome Sequencing libraries, limiting the analysis to 105 actionable genes involved in IMDs. This allows overtaking the actual limitations of NBS on DBS, enhancing our capacity to identify variants that can drive a genetic disease. Through this approach, we could reach 100% of cases solved, with 37.9% of cases (41/108) for which the combination of the biochemical and NGS analysis was indispensable for a correct diagnosis. In total, we could identify 17 affected, 34 false positives, 12 individuals referred to us for maternal conditions. In 45 newborns the molecular analysis showed heterozygosity for mutations in one or more of the genes analyzed, with results compatible with the biochemical profile indicative of NBS positivity. Conclusions In this study, we validated the performance of the proposed workflow. The advantage of this approach is limiting molecular analysis only to positive newborns and using a restricted panel of 105 genes relevant for the expanded NBS, with a 100% rate of diagnosis and potential reduction of the costs related to NBS procedures and reduced impact on patients and families.https://doi.org/10.1186/s13023-025-03546-1Newborn screening (NBS)Inherited metabolic disorders (IMDs)Whole exome sequencing (WES)Next-generation sequencing (NGS) |
| spellingShingle | Simona Fecarotta Lorenzo Vaccaro Alessandra Verde Marianna Alagia Alessandro Rossi Chiara Colantuono Maria Teresa Cacciapuoti Patrizia Annunziata Sara Riccardo Antonio Grimaldi Tonya Fusco Rosa De Santis Fernando Barretta Lucia Albano Daniela Crisci Fabiana Vallone Antonietta Tarallo Marcella Cesana Nicola Brunetti-Pierri Giulia Frisso Margherita Ruoppolo Davide Cacchiarelli Giancarlo Parenti Combined biochemical profiling and DNA sequencing in the expanded newborn screening for inherited metabolic diseases: the experience in an Italian reference center Orphanet Journal of Rare Diseases Newborn screening (NBS) Inherited metabolic disorders (IMDs) Whole exome sequencing (WES) Next-generation sequencing (NGS) |
| title | Combined biochemical profiling and DNA sequencing in the expanded newborn screening for inherited metabolic diseases: the experience in an Italian reference center |
| title_full | Combined biochemical profiling and DNA sequencing in the expanded newborn screening for inherited metabolic diseases: the experience in an Italian reference center |
| title_fullStr | Combined biochemical profiling and DNA sequencing in the expanded newborn screening for inherited metabolic diseases: the experience in an Italian reference center |
| title_full_unstemmed | Combined biochemical profiling and DNA sequencing in the expanded newborn screening for inherited metabolic diseases: the experience in an Italian reference center |
| title_short | Combined biochemical profiling and DNA sequencing in the expanded newborn screening for inherited metabolic diseases: the experience in an Italian reference center |
| title_sort | combined biochemical profiling and dna sequencing in the expanded newborn screening for inherited metabolic diseases the experience in an italian reference center |
| topic | Newborn screening (NBS) Inherited metabolic disorders (IMDs) Whole exome sequencing (WES) Next-generation sequencing (NGS) |
| url | https://doi.org/10.1186/s13023-025-03546-1 |
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