title: New tool for biological dosimetry: reevaluation and automation of the gold standard method following telomere and centromere staining.
abstract: OBJECTIVE
  The dicentric chromosome (dicentric) assay is the international gold-standard method for biological dosimetry and classification of genotoxic agents. The introduction of telomere and centromere (TC) staining offers the potential to render dicentric scoring more efficient and robust. In this study, we improved the detection of dicentrics and all unstable chromosomal aberrations (CA) leading to a significant reevaluation of the dose-effect curve and developed an automated approach following TC staining.
  
  
  METHODS
  Blood samples from 16 healthy donors were exposed to (137)Cs at 8 doses from 0.1 to 6Gy. CA were manually and automatically scored following uniform (Giemsa) or TC staining. The detection of centromeric regions and telomeric sequences using PNA probes allowed the detection of all unstable CA: dicentrics, centric and acentric rings, and all acentric fragments (with 2, 4 or no telomeres) leading to the precise quantification of estimated double strand breaks (DSB).
  
  
  RESULTS
  Manual scoring following TC staining revealed a significantly higher frequency of dicentrics (p<10(-3)) (up to 30%) and estimated DSB (p<10(-4)) compared to uniform staining due to improved detection of dicentrics with centromeres juxtaposed with other centromeres or telomeres. This improvement permitted the development of the software, TCScore, that detected 95% of manually scored dicentrics compared to 50% for the best currently available software (DCScore™).
  
  
  CONCLUSIONS
  The use of TC staining has permitted a reevaluation of the dose-response curve and the highly efficient automation of the scoring process, marking a new step in the management and follow-up of populations exposed to genotoxic agents including ionizing radiation.
authors.affiliation: Laboratoire de Radiobiologie et Oncologie (LRO), Commissariat à l'Energie Atomique (CEA), Route du Panorama, 92265 Fontenay-aux-Roses, France.
authors.forename: Radhia
authors.initials: R
authors.lastname: M'kacher
authors.affiliation: Laboratoire de Radiobiologie et Oncologie (LRO), Commissariat à l'Energie Atomique (CEA), Route du Panorama, 92265 Fontenay-aux-Roses, France; Laboratoire MIPS - Groupe TIIM3D, Université de Haute-Alsace, F-68093 Mulhouse, France.
authors.forename: Elie E L
authors.initials: EE
authors.lastname: Maalouf
authors.affiliation: Laboratoire de Radiobiologie et Oncologie (LRO), Commissariat à l'Energie Atomique (CEA), Route du Panorama, 92265 Fontenay-aux-Roses, France.
authors.forename: Michelle
authors.initials: M
authors.lastname: Ricoul
authors.affiliation: MetaSystems GmbH, Robert-Bosch-Str. 6, 68804 Altlussheim, Germany.
authors.forename: Leonhard
authors.initials: L
authors.lastname: Heidingsfelder
authors.affiliation: Pole Concept, 61 Rue Erlanger, 75016 Paris, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Laplagne
authors.affiliation: Laboratoire de Radiobiologie et Oncologie (LRO), Commissariat à l'Energie Atomique (CEA), Route du Panorama, 92265 Fontenay-aux-Roses, France.
authors.forename: Corina
authors.initials: C
authors.lastname: Cuceu
authors.affiliation: Laboratoire de Radiobiologie et Oncologie (LRO), Commissariat à l'Energie Atomique (CEA), Route du Panorama, 92265 Fontenay-aux-Roses, France.
authors.forename: William M
authors.initials: WM
authors.lastname: Hempel
authors.affiliation: Laboratoire MIPS - Groupe TIIM3D, Université de Haute-Alsace, F-68093 Mulhouse, France.
authors.forename: Bruno
authors.initials: B
authors.lastname: Colicchio
authors.affiliation: Laboratoire MIPS - Groupe TIIM3D, Université de Haute-Alsace, F-68093 Mulhouse, France.
authors.forename: Alain
authors.initials: A
authors.lastname: Dieterlen
authors.affiliation: Laboratoire de Radiobiologie et Oncologie (LRO), Commissariat à l'Energie Atomique (CEA), Route du Panorama, 92265 Fontenay-aux-Roses, France. Electronic address: laure.sabatier@cea.fr.
authors.forename: Laure
authors.initials: L
authors.lastname: Sabatier
mesh_terms: D000328:Adult; D001773:Blood Cells / Q000378:metabolism / Q000528:radiation effects; D002503:Centromere / Q000528:radiation effects; D002869:Chromosome Aberrations / Q000528:radiation effects; D004307:Dose-Response Relationship, Radiation; D001330:Electronic Data Processing; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D011839:Radiation, Ionizing; D011874:Radiometry / Q000295:instrumentation / Q000379:methods* / Q000592:standards; D013194:Staining and Labeling / Q000379:methods*; D016615:Telomere / Q000528:radiation effects; D055815:Young Adult
keywords: Biological dosimetry; Centromere staining; Dicentric; Gold standard technique; Telomere