title: Preimplantation genetic diagnosis of Marfan syndrome using multiple displacement amplification.
abstract: OBJECTIVE
  To evaluate the use of multiple displacement amplification (MDA) for whole-genome amplification in the preimplantation genetic diagnosis (PGD) of Marfan syndrome.
  
  
  METHODS
  Multiple displacement amplification was used to amplify the whole-genome directly from a single cell. The MDA product was used for polymerase chain reaction (PCR) analysis of five different loci. At this point MDA was used to develop a PGD-Marfan syndrome program.
  
  
  METHODS
  Fertility and gynecology private center in Alicante, Spain.
  
  
  METHODS
  A couple in which the husband is affected by Marfan syndrome and carries a novel mutation in the FBN-1 gene.
  
  
  METHODS
  The MDA of single cells and PCR tests for PGD.
  
  
  METHODS
  Allele drop-out (ADO), amplification efficiency rates, and the ability to detect Marfan syndrome using MDA.
  
  
  RESULTS
  We report that isothermal whole-genome amplification from single cells allowed analysis of five different loci using standard conditions. The development of a MDA-PGD protocol for Marfan syndrome allowed for the diagnosis of seven embryos. These were biopsied on day 3 of culture and analyzed. Two healthy embryos were transferred 48 hours after culture, resulting in a singleton ongoing pregnancy and the birth of a healthy child.
  
  
  CONCLUSIONS
  The MDA technique is useful for overcoming the problem of insufficient genomic DNA in PGD. The use of MDA as a universal step marks a new cycle for PGD as it allows for the diagnosis of any known gene defect by standard methods and conditions.
authors.affiliation: Bernabeu Institute of Fertility and Gynecology, Alicante, Spain. belen.lledo@ua.es
authors.forename: Belén
authors.initials: B
authors.lastname: Lledó
authors.forename: Jorge
authors.initials: J
authors.lastname: Ten
authors.forename: Francisco M
authors.initials: FM
authors.lastname: Galán
authors.forename: Rafael
authors.initials: R
authors.lastname: Bernabeu
mesh_terms: D000328:Adult; D002874:Chromosome Mapping / Q000379:methods*; D005260:Female; D020022:Genetic Predisposition to Disease / Q000235:genetics; D006801:Humans; D008297:Male; D008382:Marfan Syndrome / Q000175:diagnosis* / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D019836:Preimplantation Diagnosis / Q000379:methods*