title: Improving FISH diagnosis for preimplantation genetic aneuploidy screening.
abstract: BACKGROUND
  In our routine programme of preimplantation genetic aneuploidy screening (PGS) by fluorescence in situ hybridization (FISH), nine chromosomes (13, 15, 16, 17, 18, 21, 22, X and Y) are analysed in two consecutive hybridization rounds. We also perform additional hybridization rounds for these chromosomes, using probes that bind to different loci, for non-conclusive results and for confirmation of certain aneuploidies. The aim of this study was to evaluate the impact of additional hybridization rounds on FISH accuracy.
  
  
  METHODS
  This is a retrospective analysis of our FISH data from 1000 PGS cycles performed from December 2007 to December 2008 for various indications. In addition to the hybridization rounds described above, 132 of the embryos diagnosed as chromosomally abnormal were re-analysed on Day 5.
  
  
  RESULTS
  A total of 2477 embryos were re-hybridized, 1496 due to non-conclusive results and 981 to confirm observed aneuploidies. After re-hybridization, 882 embryos (59%) were then diagnosed as normal, 600 embryos (40.1%) had a clear abnormality and only 14 embryos (0.9%) remained non-informative. From the 981 embryos in the latter group, 890 embryos had monosomies and, after re-hybridization 174 embryos (19.6%) were normal and 716 (80.5%) had confirmed monosomies. In contrast, re-hybridization confirmed 90 (98.9%) of the 91 observed trisomies. In addition, Day-5 re-analysis of abnormal embryos showed a higher rate of concordant diagnosis between Day 3 and Day 5 when re-hybridizations had been included on Day-3 (95 versus 82.7%; P= 0.0443), especially for the confirmation of monosomies (82.8 versus 61.0%; P = 0.0087).
  
  
  CONCLUSIONS
  Our data indicate that additional hybridization rounds improve the accuracy of the diagnosis, increasing the number of chromosomally normal embryos available for transfer. Re-hybridization with additional probes as a standard approach to PGS could enhance the potential benefits of the technique.
authors.affiliation: PGD Unit, Institut Universitari IVI-Valencia, Plaza Policía Local, 3, 46015 Valencia, Spain. pmir@ivi.es
authors.forename: P
authors.initials: P
authors.lastname: Mir
authors.forename: L
authors.initials: L
authors.lastname: Rodrigo
authors.forename: E
authors.initials: E
authors.lastname: Mateu
authors.forename: V
authors.initials: V
authors.lastname: Peinado
authors.forename: M
authors.initials: M
authors.lastname: Milán
authors.forename: A
authors.initials: A
authors.lastname: Mercader
authors.forename: P
authors.initials: P
authors.lastname: Buendía
authors.forename: A
authors.initials: A
authors.lastname: Delgado
authors.forename: A
authors.initials: A
authors.lastname: Pellicer
authors.forename: J
authors.initials: J
authors.lastname: Remohí
authors.forename: C
authors.initials: C
authors.lastname: Rubio
mesh_terms: D000782:Aneuploidy; D001755:Blastocyst / Q000648:ultrastructure*; D025063:Chromosome Disorders / Q000175:diagnosis*; D002877:Chromosomes, Human; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D019836:Preimplantation Diagnosis / Q000379:methods*; D012189:Retrospective Studies; D012680:Sensitivity and Specificity