title: Dual blastomere analysis improves reliability of preimplantation trembler mouse diagnosis.
abstract: Dual blastomere biopsy and independent blastomere analysis dramatically improved preimplantation diagnostic reliability as confirmed by testing the remaining biopsied eight-cell mouse embryo. The autosomal dominant trembler mouse point mutation was selected as a model for human preimplantation diagnosis because: (1) single cell assay failure is predicted to be the highest when testing autosomal dominant mutations; (2) point mutations represent the most common of all mutation categories and the most demanding mutation to assay reliably; and (3) the trembler mouse point mutation in peripheral myelin protein 22 (Pmp22) is a model of human Charcot-Marie-Tooth type 1A disease. Mathematical models predict our experimental results assuming amplification of 80% of each target allele as well as trembler sperm DNA contamination in 1 of 44 normal biopsied single blastomeres. Single blastomere analysis correctly predicted the genotype in only 84% of embryos that would have been implanted as normal. In contrast, when independent tests of both biopsied blastomeres agreed, test results were confirmed in 20 of 21 (95.2%) of the remaining six-cell biopsied embryos designated as normal. Thus, biopsied six-cell embryo confirmation demonstrated that dual biopsied blastomere analysis improved test reliability remarkably.
authors.affiliation: Departments of Obstetrics, Gynecology, and Reproductive Sciences, University of California, San Francisco 94143-0720, USA.
authors.forename: H
authors.initials: H
authors.lastname: Sago
authors.forename: H S
authors.initials: HS
authors.lastname: Kim
authors.forename: J D
authors.initials: JD
authors.lastname: Goldberg
authors.forename: J H
authors.initials: JH
authors.lastname: Chung
authors.forename: R A
authors.initials: RA
authors.lastname: Pedersen
authors.forename: R V
authors.initials: RV
authors.lastname: Lebo
mesh_terms: D000818:Animals; D001706:Biopsy; D001757:Blastomeres; D002607:Charcot-Marie-Tooth Disease / Q000175:diagnosis*; D047108:Embryonic Development; D005260:Female; D051379:Mice; D008818:Mice, Neurologic Mutants; D009185:Myelin Proteins / Q000235:genetics*; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*; D015203:Reproducibility of Results
chemical_list: D009185:Myelin Proteins; C489545:Pmp22 protein, mouse