Second-Tier Next Generation Sequencing Integrated in Nationwide Newborn Screening Provides Rapid Molecular Diagnostics of Severe Combined Immunodeficiency
Severe combined immunodeficiency (SCID) and other T cell lymphopenias can be detected during newborn screening (NBS) by measuring T cell receptor excision circles (TRECs) in dried blood spot (DBS) DNA. Second tier next generation sequencing (NGS) with an amplicon based targeted gene panel using the...
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Frontiers Media S.A.
2020-07-01
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Online Access: | https://www.frontiersin.org/article/10.3389/fimmu.2020.01417/full |
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English |
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author |
Janne Strand Kiran Aftab Gul Hans Christian Erichsen Hans Christian Erichsen Emma Lundman Mona C. Berge Anette K. Trømborg Linda K. Sørgjerd Mari Ytre-Arne Silje Hogner Ruth Halsne Ruth Halsne Hege Junita Gaup Liv T. Osnes Grete A. B. Kro Hanne S. Sorte Lars Mørkrid Alexander D. Rowe Trine Tangeraas Trine Tangeraas Jens V. Jørgensen Jens V. Jørgensen Charlotte Alme Trude E. H. Bjørndalen Arild E. Rønnestad Astri M. Lang Terje Rootwelt Terje Rootwelt Jochen Buechner Torstein Øverland Tore G. Abrahamsen Tore G. Abrahamsen Rolf D. Pettersen Asbjørg Stray-Pedersen Asbjørg Stray-Pedersen |
spellingShingle |
Janne Strand Kiran Aftab Gul Hans Christian Erichsen Hans Christian Erichsen Emma Lundman Mona C. Berge Anette K. Trømborg Linda K. Sørgjerd Mari Ytre-Arne Silje Hogner Ruth Halsne Ruth Halsne Hege Junita Gaup Liv T. Osnes Grete A. B. Kro Hanne S. Sorte Lars Mørkrid Alexander D. Rowe Trine Tangeraas Trine Tangeraas Jens V. Jørgensen Jens V. Jørgensen Charlotte Alme Trude E. H. Bjørndalen Arild E. Rønnestad Astri M. Lang Terje Rootwelt Terje Rootwelt Jochen Buechner Torstein Øverland Tore G. Abrahamsen Tore G. Abrahamsen Rolf D. Pettersen Asbjørg Stray-Pedersen Asbjørg Stray-Pedersen Second-Tier Next Generation Sequencing Integrated in Nationwide Newborn Screening Provides Rapid Molecular Diagnostics of Severe Combined Immunodeficiency Frontiers in Immunology SCID - severe combined immunodeficiency newborn SCID screening NGS - next generation sequencing severe T-cell immunodeficiency TREC analysis |
author_facet |
Janne Strand Kiran Aftab Gul Hans Christian Erichsen Hans Christian Erichsen Emma Lundman Mona C. Berge Anette K. Trømborg Linda K. Sørgjerd Mari Ytre-Arne Silje Hogner Ruth Halsne Ruth Halsne Hege Junita Gaup Liv T. Osnes Grete A. B. Kro Hanne S. Sorte Lars Mørkrid Alexander D. Rowe Trine Tangeraas Trine Tangeraas Jens V. Jørgensen Jens V. Jørgensen Charlotte Alme Trude E. H. Bjørndalen Arild E. Rønnestad Astri M. Lang Terje Rootwelt Terje Rootwelt Jochen Buechner Torstein Øverland Tore G. Abrahamsen Tore G. Abrahamsen Rolf D. Pettersen Asbjørg Stray-Pedersen Asbjørg Stray-Pedersen |
author_sort |
Janne Strand |
title |
Second-Tier Next Generation Sequencing Integrated in Nationwide Newborn Screening Provides Rapid Molecular Diagnostics of Severe Combined Immunodeficiency |
title_short |
Second-Tier Next Generation Sequencing Integrated in Nationwide Newborn Screening Provides Rapid Molecular Diagnostics of Severe Combined Immunodeficiency |
title_full |
Second-Tier Next Generation Sequencing Integrated in Nationwide Newborn Screening Provides Rapid Molecular Diagnostics of Severe Combined Immunodeficiency |
title_fullStr |
Second-Tier Next Generation Sequencing Integrated in Nationwide Newborn Screening Provides Rapid Molecular Diagnostics of Severe Combined Immunodeficiency |
title_full_unstemmed |
Second-Tier Next Generation Sequencing Integrated in Nationwide Newborn Screening Provides Rapid Molecular Diagnostics of Severe Combined Immunodeficiency |
title_sort |
second-tier next generation sequencing integrated in nationwide newborn screening provides rapid molecular diagnostics of severe combined immunodeficiency |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Immunology |
issn |
1664-3224 |
publishDate |
2020-07-01 |
description |
Severe combined immunodeficiency (SCID) and other T cell lymphopenias can be detected during newborn screening (NBS) by measuring T cell receptor excision circles (TRECs) in dried blood spot (DBS) DNA. Second tier next generation sequencing (NGS) with an amplicon based targeted gene panel using the same DBS DNA was introduced as part of our prospective pilot research project in 2015. With written parental consent, 21 000 newborns were TREC-tested in the pilot. Three newborns were identified with SCID, and disease-causing variants in IL2RG, RAG2, and RMRP were confirmed by NGS on the initial DBS DNA. The molecular findings directed follow-up and therapy: the IL2RG-SCID underwent early hematopoietic stem cell transplantation (HSCT) without any complications; the leaky RAG2-SCID received prophylactic antibiotics, antifungals, and immunoglobulin infusions, and underwent HSCT at 1 year of age. The child with RMRP-SCID had complete Hirschsprung disease and died at 1 month of age. Since January 2018, all newborns in Norway have been offered NBS for SCID using 1st tier TRECs and 2nd tier gene panel NGS on DBS DNA. During the first 20 months of nationwide SCID screening an additional 88 000 newborns were TREC tested, and four new SCID cases were identified. Disease-causing variants in DCLRE1C, JAK3, NBN, and IL2RG were molecularly confirmed on day 8, 15, 8 and 6, respectively after birth, using the initial NBS blood spot. Targeted gene panel NGS integrated into the NBS algorithm rapidly delineated the specific molecular diagnoses and provided information useful for management, targeted therapy and follow-up i.e., X rays and CT scans were avoided in the radiosensitive SCID. Second tier targeted NGS on the same DBS DNA as the TREC test provided instant confirmation or exclusion of SCID, and made it possible to use a less stringent TREC cut-off value. This allowed for the detection of leaky SCIDs, and simultaneously reduced the number of control samples, recalls and false positives. Mothers were instructed to stop breastfeeding until maternal cytomegalovirus (CMV) status was determined. Our limited data suggest that shorter time-interval from birth to intervention, may prevent breast milk transmitted CMV infection in classical SCID. |
topic |
SCID - severe combined immunodeficiency newborn SCID screening NGS - next generation sequencing severe T-cell immunodeficiency TREC analysis |
url |
https://www.frontiersin.org/article/10.3389/fimmu.2020.01417/full |
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doaj-634d7c285f864c7781322eb1360b861b2020-11-25T03:52:32ZengFrontiers Media S.A.Frontiers in Immunology1664-32242020-07-011110.3389/fimmu.2020.01417545364Second-Tier Next Generation Sequencing Integrated in Nationwide Newborn Screening Provides Rapid Molecular Diagnostics of Severe Combined ImmunodeficiencyJanne Strand0Kiran Aftab Gul1Hans Christian Erichsen2Hans Christian Erichsen3Emma Lundman4Mona C. Berge5Anette K. Trømborg6Linda K. Sørgjerd7Mari Ytre-Arne8Silje Hogner9Ruth Halsne10Ruth Halsne11Hege Junita Gaup12Liv T. Osnes13Grete A. B. Kro14Hanne S. Sorte15Lars Mørkrid16Alexander D. Rowe17Trine Tangeraas18Trine Tangeraas19Jens V. Jørgensen20Jens V. Jørgensen21Charlotte Alme22Trude E. H. Bjørndalen23Arild E. Rønnestad24Astri M. Lang25Terje Rootwelt26Terje Rootwelt27Jochen Buechner28Torstein Øverland29Tore G. Abrahamsen30Tore G. Abrahamsen31Rolf D. Pettersen32Asbjørg Stray-Pedersen33Asbjørg Stray-Pedersen34Norwegian National Unit for Newborn Screening, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwayPaediatric Research Institute, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwayDepartment of Paediatrics, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwayDivision of Paediatric and Adolescent Medicine, Institute of Clinical Medicine, University of Oslo, Oslo, NorwayNorwegian National Unit for Newborn Screening, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwayNorwegian National Unit for Newborn Screening, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwayNorwegian National Unit for Newborn Screening, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwayNorwegian National Unit for Newborn Screening, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwayNorwegian National Unit for Newborn Screening, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwayNorwegian National Unit for Newborn Screening, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwayNorwegian National Unit for Newborn Screening, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwayDepartment of Forensic Biology, Oslo University Hospital, Oslo, NorwayNorwegian National Unit for Newborn Screening, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwayDepartment of Immunology and Transfusion Medicine, Oslo University Hospital, Oslo, NorwayDepartment of Microbiology, Oslo University Hospital, Oslo, NorwayDepartment of Medical Genetics, Oslo University Hospital, Oslo, NorwayDepartment of Medical Biochemistry, Oslo University Hospital, Oslo, NorwayNorwegian National Unit for Newborn Screening, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwayNorwegian National Unit for Newborn Screening, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwayDepartment of Paediatrics, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwayNorwegian National Unit for Newborn Screening, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwayDepartment of Paediatrics, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, Norway0Department of Paediatric Haematology, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, Norway1Department of Obstetrics and Gynaecology, Oslo University Hospital, Oslo, NorwayDepartment of Paediatrics, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwayDepartment of Paediatrics, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwayDepartment of Paediatrics, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwayDivision of Paediatric and Adolescent Medicine, Institute of Clinical Medicine, University of Oslo, Oslo, Norway0Department of Paediatric Haematology, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwayDepartment of Paediatrics, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwayDepartment of Paediatrics, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwayDivision of Paediatric and Adolescent Medicine, Institute of Clinical Medicine, University of Oslo, Oslo, NorwayNorwegian National Unit for Newborn Screening, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwayNorwegian National Unit for Newborn Screening, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwayDepartment of Paediatrics, Division of Paediatric and Adolescent Medicine, Oslo University Hospital, Oslo, NorwaySevere combined immunodeficiency (SCID) and other T cell lymphopenias can be detected during newborn screening (NBS) by measuring T cell receptor excision circles (TRECs) in dried blood spot (DBS) DNA. Second tier next generation sequencing (NGS) with an amplicon based targeted gene panel using the same DBS DNA was introduced as part of our prospective pilot research project in 2015. With written parental consent, 21 000 newborns were TREC-tested in the pilot. Three newborns were identified with SCID, and disease-causing variants in IL2RG, RAG2, and RMRP were confirmed by NGS on the initial DBS DNA. The molecular findings directed follow-up and therapy: the IL2RG-SCID underwent early hematopoietic stem cell transplantation (HSCT) without any complications; the leaky RAG2-SCID received prophylactic antibiotics, antifungals, and immunoglobulin infusions, and underwent HSCT at 1 year of age. The child with RMRP-SCID had complete Hirschsprung disease and died at 1 month of age. Since January 2018, all newborns in Norway have been offered NBS for SCID using 1st tier TRECs and 2nd tier gene panel NGS on DBS DNA. During the first 20 months of nationwide SCID screening an additional 88 000 newborns were TREC tested, and four new SCID cases were identified. Disease-causing variants in DCLRE1C, JAK3, NBN, and IL2RG were molecularly confirmed on day 8, 15, 8 and 6, respectively after birth, using the initial NBS blood spot. Targeted gene panel NGS integrated into the NBS algorithm rapidly delineated the specific molecular diagnoses and provided information useful for management, targeted therapy and follow-up i.e., X rays and CT scans were avoided in the radiosensitive SCID. Second tier targeted NGS on the same DBS DNA as the TREC test provided instant confirmation or exclusion of SCID, and made it possible to use a less stringent TREC cut-off value. This allowed for the detection of leaky SCIDs, and simultaneously reduced the number of control samples, recalls and false positives. Mothers were instructed to stop breastfeeding until maternal cytomegalovirus (CMV) status was determined. Our limited data suggest that shorter time-interval from birth to intervention, may prevent breast milk transmitted CMV infection in classical SCID.https://www.frontiersin.org/article/10.3389/fimmu.2020.01417/fullSCID - severe combined immunodeficiencynewborn SCID screeningNGS - next generation sequencingsevere T-cell immunodeficiencyTREC analysis |