title: Familial haplotyping and embryo analysis for Preimplantation genetic diagnosis (PGD) using DNA microarrays: a proof of principle study.
abstract: OBJECTIVE
  Development of PGD assays for molecular disorders is based on analysis of a familial mutation together with linked polymorphic STR markers; a process which is lengthy and requires the identification of multiple informative markers prior to PGD analysis. On the other hand, whole genome amplification (WGA), in conjunction with microarray platforms, allows the use of a universal assay for the analysis of a very large number of SNP markers at once. The aim of this study was to test high throughput pre-PGD familial haplotyping for in-case blastomere analysis in order to eliminate time-consuming pre-case preparations for each family.
  
  
  METHODS
  A PGD cycle was performed for a couple with paternal Charcot Marie Tooth 1A (CMT1A) using a classic multiplex nested PCR approach. Mutant embryos from the case were blindly reanalyzed, as single or multi-cell biopsies, using a multiple displacement amplification-based WGA protocol and microarray SNP analysis. In parallel, relevant genomic DNA samples from the family were also analyzed by SNP microarray.
  
  
  RESULTS
  After applying a 'unique informative allele' selection algorithm to the data, this array-based assay reconfirmed the initial diagnosis in all samples.
  
  
  CONCLUSIONS
  We describe a PGD method that is both accurate and feasible during the time-frame required for embryo transfer. This strategy greatly reduces the time for pre-case haplotype preparation.
authors.affiliation: Medical Genetics Institute, ZOHAR PGD Lab and IVF Unit, Shaare Zedek Medical Center, Jerusalem, Israel, 91031, gheona@szmc.org.il.
authors.forename: G
authors.initials: G
authors.lastname: Altarescu
authors.forename: D A
authors.initials: DA
authors.lastname: Zeevi
authors.forename: S
authors.initials: S
authors.lastname: Zeligson
authors.forename: S
authors.initials: S
authors.lastname: Perlberg
authors.forename: T
authors.initials: T
authors.lastname: Eldar-Geva
authors.forename: E J
authors.initials: EJ
authors.lastname: Margalioth
authors.forename: E
authors.initials: E
authors.lastname: Levy-Lahad
authors.forename: P
authors.initials: P
authors.lastname: Renbaum
mesh_terms: D000483:Alleles; D001706:Biopsy; D002607:Charcot-Marie-Tooth Disease / Q000175:diagnosis / Q000235:genetics; D004624:Embryo Transfer; D005260:Female; D005784:Gene Amplification; D030342:Genetic Diseases, Inborn / Q000175:diagnosis* / Q000235:genetics; D006239:Haplotypes / Q000235:genetics*; D006801:Humans; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D011247:Pregnancy; D019836:Preimplantation Diagnosis / Q000379:methods*