title: Cystic Fibrosis Mutation Detection with SPR Biosensor in Real Samples via Multiple Surfaces Binding Method.
abstract: OBJECTIVE
  Surface Plasmon Resonance (SPR) based biosensor system was developed for the detection of Delta F508 (ΔF508del) Cystic Fibrosis (CF) mutation in both synthetic and real samples.
  
  
  METHODS
  In order to provide an effective hybridization between probe and the Polymerase Chain Reaction (PCR) amplicons (target), streptavidin was bound to the surface and biotin-tag probe was sent to the streptavidin-coated surface. For the target preparation, blood samples were collected from the patients who suffer from CF. Following the DNA isolation; samples were amplified with PCR with biotin-tag. Before sending the biotin-tag PCR amplicons onto the modified surface, amplicons were also interacted with the helper oligonucleotides to prevent re-annealing of the denatured DNA strands. This kind of 'multiple surface binding' method helps increasing the sensitivity of the detection.
  
  
  RESULTS
  The limit of detection (S/N= 3) was calculated as 12.24 pico-mole/ml for PCR-like synthetic long target sequence and 13x105 molecules for real samples in less than half an hour.
  
  
  CONCLUSIONS
  Using the both biotin-tag probe and the helper oligonucleotides together, hybridization was achieved much more efficiently than traditional denaturation protocols for real samples and biotinfree hybridization detection. To the best of our knowledge, the procedure described in this study is one of the simplest, rapid and sensitive methods for CF mutation detection with SPR based biosensor system in real samples.
authors.affiliation: Department of Analytical Chemistry, Faculty of Pharmacy, Ege University, Izmir. Turkey.
authors.forename: Yasin Ugur
authors.initials: YU
authors.lastname: Kayran
authors.affiliation: Department of Analytical Chemistry, Faculty of Pharmacy, Ege University, Izmir. Turkey.
authors.forename: Dilsat
authors.initials: D
authors.lastname: Ozkan-Ariksoysal
authors.affiliation: Department of Analytical Chemistry, Faculty of Pharmacy, Ege University, Izmir. Turkey.
authors.forename: Seda Nur
authors.initials: SN
authors.lastname: Topkaya
authors.affiliation: Department of Medicinal Biology, Faculty of Medicine, Ege University, Izmir. Turkey.
authors.forename: Burcin Tezcanli
authors.initials: BT
authors.lastname: Kaymaz
authors.affiliation: Department of Medicinal Biology, Faculty of Medicine, Ege University, Izmir. Turkey.
authors.forename: Buket Kosova
authors.initials: BK
authors.lastname: Can
mesh_terms: D020137:Base Pair Mismatch; D003550:Cystic Fibrosis / Q000235:genetics*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D004247:DNA / Q000235:genetics*; D006801:Humans; D009154:Mutation; D009693:Nucleic Acid Hybridization / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: C093349:cystic fibrosis transmembrane conductance regulator delta F508; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004247:DNA
keywords: DNA
  hybridization; Delta F508 (ΔF508del) cystic fibrosis mutation; Surface Plasmon Resonance (SPR) spectroscopy; optical biosensor; polymerase chain reaction (PCR) amplified samples