title: Improving newborn screening for cystic fibrosis using next-generation sequencing technology: a technical feasibility study.
abstract: OBJECTIVE
  Many regions have implemented newborn screening (NBS) for cystic fibrosis (CF) using a limited panel of cystic fibrosis transmembrane regulator (CFTR) mutations after immunoreactive trypsinogen (IRT) analysis. We sought to assess the feasibility of further improving the screening using next-generation sequencing (NGS) technology.
  
  
  METHODS
  An NGS assay was used to detect 162 CFTR mutations/variants characterized by the CFTR2 project. We used 67 dried blood spots (DBSs) containing 48 distinct CFTR mutations to validate the assay. NGS assay was retrospectively performed on 165 CF screen-positive samples with one CFTR mutation.
  
  
  RESULTS
  The NGS assay was successfully performed using DNA isolated from DBSs, and it correctly detected all CFTR mutations in the validation. Among 165 screen-positive infants with one CFTR mutation, no additional disease-causing mutation was identified in 151 samples consistent with normal sweat tests. Five infants had a CF-causing mutation that was not included in this panel, and nine with two CF-causing mutations were identified.
  
  
  CONCLUSIONS
  The NGS assay was 100% concordant with traditional methods. Retrospective analysis results indicate an IRT/NGS screening algorithm would enable high sensitivity, better specificity and positive predictive value (PPV). This study lays the foundation for prospective studies and for introducing NGS in NBS laboratories.
authors: Department of Pediatrics, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin, USA.|Newborn Screening Laboratory, Wisconsin State Laboratory of Hygiene, University of Wisconsin-Madison, Madison, Wisconsin, USA.
authors: Mei W
authors: http://orcid.org/0000-0002-8707-1834
authors: MW
authors: Baker
authors: Newborn Screening Laboratory, Wisconsin State Laboratory of Hygiene, University of Wisconsin-Madison, Madison, Wisconsin, USA.
authors: Anne E
authors: AE
authors: Atkins
authors: The Newborn Screening and Molecular Biology Branch, Centers for Disease Control and Prevention, Atlanta, Georgia, USA.
authors: Suzanne K
authors: SK
authors: Cordovado
authors: The Newborn Screening and Molecular Biology Branch, Centers for Disease Control and Prevention, Atlanta, Georgia, USA.
authors: Miyono
authors: M
authors: Hendrix
authors: The Newborn Screening and Molecular Biology Branch, Centers for Disease Control and Prevention, Atlanta, Georgia, USA.
authors: Marie C
authors: MC
authors: Earley
authors: Department of Pediatrics, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin, USA.|Department of Population Health Sciences, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin, USA.
authors: Philip M
authors: PM
authors: Farrell
mesh_terms: D003550:Cystic Fibrosis / Q000175:diagnosis* / Q000235:genetics; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D059788:Dried Blood Spot Testing; D005240:Feasibility Studies; D020022:Genetic Predisposition to Disease; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D007231:Infant, Newborn; D009154:Mutation; D015997:Neonatal Screening / Q000379:methods*; D012189:Retrospective Studies; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: C505032:CFTR protein, human; D019005:Cystic Fibrosis Transmembrane Conductance Regulator