title: Cost-effective and scalable DNA extraction method from dried blood spots.
abstract: BACKGROUND
  Dried blood spot (DBS) samples have been widely used in newborn screening (NBS) for the early identification of disease to facilitate the presymptomatic treatment of congenital diseases in newborns. As molecular genetics knowledge and technology progresses, there is an increased demand on NBS programs for molecular testing and a need to establish reliable, low-cost methods to perform those analyses. Here we report a flexible, cost-efficient, high-throughput DNA extraction method from DBS adaptable to small- and large-scale screening settings.
  
  
  METHODS
  Genomic DNA (g.DNA) was extracted from single 3-mm diameter DBS by the sequential use of red cell lysis, detergent-alkaline, and acid-neutralizing buffers routinely used in whole blood and plant tissue DNA extractions. We performed PCR amplification of several genomic regions using standard PCR conditions and detection methods (agarose gel, melting-curve analysis, TaqMan-based assays). Amplicons were confirmed by BigDye® Terminator cycle sequencing and compared with reference sequences.
  
  
  RESULTS
  High-quality g.DNA was extracted from hundreds of DBS, as proven by mutation detection of several human genes on multiple platforms. Manual and automated extraction protocols were validated. Quantification of g.DNA by Oligreen® fluorescent nucleic acid stain demonstrated a normal population distribution closely corresponding with white blood cell counts detected in newborn populations.
  
  
  CONCLUSIONS
  High-quality, amplifiable g.DNA is extractable from DBSs. Our method is adaptable, reliable, and scalable to low- and high-throughput NBS at low cost ($0.10/sample). This method is routinely used for molecular testing in the New York State NBS program.
authors.affiliation: Newborn Screening Program, Division of Genetics, Wadsworth Center, New York State Department of Health, Albany, NY 12201, USA. saavedra@wadsworth.org
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Saavedra-Matiz
authors.forename: Jason T
authors.initials: JT
authors.lastname: Isabelle
authors.forename: Chad K
authors.initials: CK
authors.lastname: Biski
authors.forename: Salvatore J
authors.initials: SJ
authors.lastname: Duva
authors.forename: Melissa L
authors.initials: ML
authors.lastname: Sweeney
authors.forename: April L
authors.initials: AL
authors.lastname: Parker
authors.forename: Allison J
authors.initials: AJ
authors.lastname: Young
authors.forename: Lisa L
authors.initials: LL
authors.lastname: Diantonio
authors.forename: Lea M
authors.initials: LM
authors.lastname: Krein
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Nichols
authors.forename: Michele
authors.initials: M
authors.lastname: Caggana
mesh_terms: D003362:Cost-Benefit Analysis; D004247:DNA / Q000097:blood / Q000302:isolation & purification*; D059788:Dried Blood Spot Testing / Q000191:economics / Q000379:methods*; D006801:Humans; D007231:Infant, Newborn; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D004247:DNA