title: Preimplantation diagnosis of single gene disorders by two-step oocyte genetic analysis using first and second polar body.
abstract: Previous work on preimplantation genetic diagnosis (PGD) of single gene disorders by the first polar body (IPB) analysis has demonstrated that the genotype of a considerable number of embryos resulting from heterozygous oocytes cannot be predicted without testing their second PB (IIPB). To overcome this limitation we introduce a two-step DNA analysis of oocytes using both IPB and IIPB to identify hemizygous mutation-free oocytes following the second meiotic division. In the application of the approach to PGD of cystic fibrosis (CF) Delta F-508 mutation, sickle cell disease, and hemophilia B, 80 oocytes were studied by both PBs, resulting in the identification and transfer of 32 homozygous normal embryos. A follow-up genotyping of 52 embryos, resulting from oocytes tested by both IPB and IIPB demonstrated the accuracy of the predicted genotypes. In addition to a nested PCR analysis of the mutant genes in PBs and resulting embryos, simultaneous amplification of different polymorphic markers was performed, demonstrating the reliability of the two-step polar body analysis of oocytes.
authors.affiliation: Reproductive Genetics Institute, Illinois Masonic Medical Center, Chicago 60657, USA.
authors.forename: Y
authors.initials: Y
authors.lastname: Verlinsky
authors.forename: S
authors.initials: S
authors.lastname: Rechitsky
authors.forename: J
authors.initials: J
authors.lastname: Cieslak
authors.forename: V
authors.initials: V
authors.lastname: Ivakhnenko
authors.forename: G
authors.initials: G
authors.lastname: Wolf
authors.forename: A
authors.initials: A
authors.lastname: Lifchez
authors.forename: B
authors.initials: B
authors.lastname: Kaplan
authors.forename: J
authors.initials: J
authors.lastname: Moise
authors.forename: J
authors.initials: J
authors.lastname: Walle
authors.forename: M
authors.initials: M
authors.lastname: White
authors.forename: N
authors.initials: N
authors.lastname: Ginsberg
authors.forename: C
authors.initials: C
authors.lastname: Strom
authors.forename: A
authors.initials: A
authors.lastname: Kuliev
mesh_terms: D000483:Alleles; D004247:DNA / Q000032:analysis; D004622:Embryo, Mammalian / Q000737:chemistry; D005260:Female; D005307:Fertilization in Vitro; D005500:Follow-Up Studies; D030342:Genetic Diseases, Inborn / Q000175:diagnosis* / Q000235:genetics* / Q000517:prevention & control; D005838:Genotype; D006801:Humans; D008540:Meiosis / Q000235:genetics; D009865:Oocytes / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D019836:Preimplantation Diagnosis / Q000379:methods*
chemical_list: D004247:DNA