title: Newborn screening for severe combined immunodeficiency using a novel and simplified method to measure T-cell excision circles (TREC).
abstract: The prognosis of children with severe combined immunodeficiency (SCID) depends on a presymptomatic diagnosis and early treatment before complications occur. We established and tested a simplified, practical and economic newborn screening method based on the quantification of T-cell receptor excision circles (TRECs) on dried blood spots (DBSs) through qPCR. Our method was validated by the analysis of 11 positive controls, which all showed an absence of TRECs, thus yielding a sensitivity of 100%. Further, we analyzed 6034 anonymized newborns of whom 6031 (99,95%) showed a normal TREC qPCR with a median of 600 estimated TREC copies/1.6mm punch. The test showed a recall-rate of 0.05%. We present a highly sensitive, specific and time- and cost-effective method of TREC quantification, which is suitable for SCID newborn screening. In comparison to established methods, our test requires only 25% of the input material, doesn't require DNA purification and significantly reduces time and cost requirement.
authors.affiliation: Department of Pediatric Oncology, Hematology and Immunology, Children's Hospital, University of Heidelberg, Heidelberg, Germany; Pediatric Highly Intensive Care Unit, Department of Pathophysiology and Transplantation, Università degli Studi di Milano, Fondazione IRCCS Ca'randa, Ospedale Maggiore Policlinico, Milan, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Tagliaferri
authors.affiliation: Department of Pediatric Oncology, Hematology and Immunology, Children's Hospital, University of Heidelberg, Heidelberg, Germany.
authors.forename: Joachim B
authors.initials: JB
authors.lastname: Kunz
authors.affiliation: Department of Pediatric Oncology, Hematology and Immunology, Children's Hospital, University of Heidelberg, Heidelberg, Germany.
authors.forename: Margit
authors.initials: M
authors.lastname: Happich
authors.affiliation: Pediatric Highly Intensive Care Unit, Department of Pathophysiology and Transplantation, Università degli Studi di Milano, Fondazione IRCCS Ca'randa, Ospedale Maggiore Policlinico, Milan, Italy.
authors.forename: Susanna
authors.initials: S
authors.lastname: Esposito
authors.affiliation: Institute of Medical Biometry and Informatics, University of Heidelberg, Heidelberg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Bruckner
authors.affiliation: Department of Pediatric Oncology, Hematology and Immunology, Children's Hospital, University of Heidelberg, Heidelberg, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Hübschmann
authors.affiliation: Division of Metabolic Diseases and Newborn Screening Center, Department of Paediatrics I, Children's Hospital, University of Heidelberg, Heidelberg, Germany.
authors.forename: Jürgen G
authors.initials: JG
authors.lastname: Okun
authors.affiliation: Division of Metabolic Diseases and Newborn Screening Center, Department of Paediatrics I, Children's Hospital, University of Heidelberg, Heidelberg, Germany.
authors.forename: Georg F
authors.initials: GF
authors.lastname: Hoffmann
authors.affiliation: Department of Pediatrics, University Medical Center Ulm, Germany.
authors.forename: Ansgar
authors.initials: A
authors.lastname: Schulz
authors.affiliation: Department of Pediatrics, University Medical Center Ulm, Germany.
authors.forename: Judit
authors.initials: J
authors.lastname: Kappe
authors.affiliation: Center for Pediatrics and Adolescent Medicine and Center of Chronic Immunodeficiency, University Medical Center Freiburg, Germany.
authors.forename: Carsten
authors.initials: C
authors.lastname: Speckmann
authors.affiliation: Department of Pediatric Oncology, Hematology and Immunology, Children's Hospital, University of Heidelberg, Heidelberg, Germany.
authors.forename: Martina U
authors.initials: MU
authors.lastname: Muckenthaler
authors.affiliation: Department of Pediatric Oncology, Hematology and Immunology, Children's Hospital, University of Heidelberg, Heidelberg, Germany. Electronic address: andreas.kulozik@med.uni-heidelberg.de.
authors.forename: Andreas E
authors.initials: AE
authors.lastname: Kulozik
mesh_terms: D004247:DNA / Q000235:genetics; D059788:Dried Blood Spot Testing / Q000379:methods; D006801:Humans; D007231:Infant, Newborn; D015997:Neonatal Screening / Q000379:methods*; D010865:Pilot Projects; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000276:immunology; D016511:Severe Combined Immunodeficiency / Q000175:diagnosis* / Q000235:genetics / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D011948:Receptors, Antigen, T-Cell; D004247:DNA
keywords: Newborn screening; Severe combined immunodeficiency; T-cell receptor excision circles (TRECs)