title: Carrier-specific breakpoint-spanning DNA probes: an approach to preimplantation genetic diagnosis in interphase cells.
abstract: Carriers of chromosomal inversions or other balanced rearrangements represent a significant fraction of patients in in-vitro fertilization (IVF) programmes due to recurrent reproductive problems. In most cases, chromosomal imbalance in fertilized oocytes is incompatible with embryo survival leading to increased rates of spontaneous abortions. Assuming that a fraction of the germ cells is karyotypically normal, these patients would greatly benefit from efficient procedures for generation and use of breakpoint-specific DNA hybridization probes in preconception and preimplantation genetic diagnosis (PGD). We describe the generation of such patient-specific probes to discriminate between normal and aberrant chromosomes in interphase cells. First, a large insert DNA library was screened for probes that bind adjacent to the chromosomal breakpoints or span them. Then, probe and hybridization parameters were optimized using white blood cells from the carrier to increase in hybridization signal intensity and contrast. Finally, the probes were tested on target cells (typically polar bodies or blastomeres) and a decision about the colour labelling scheme was made, before the probes can be used for preconception or preimplantation genetic analysis. Thus, it was demonstrated that cells with known structural abnormalities could be detected, based on hybridization of breakpoint spanning yeast artificial chromosome (YAC) DNA probes in interphase cells.
authors.affiliation: Resource for Molecular Cytogenetics, Life Sciences Division, University of California, Berkeley 94720, USA.
authors.forename: M J
authors.initials: MJ
authors.lastname: Cassel
authors.forename: S
authors.initials: S
authors.lastname: Munné
authors.forename: J
authors.initials: J
authors.lastname: Fung
authors.forename: H U
authors.initials: HU
authors.lastname: Weier
mesh_terms: D001757:Blastomeres / Q000737:chemistry; D002869:Chromosome Aberrations; D002873:Chromosome Fragility; D007446:Chromosome Inversion; D018244:Chromosomes, Artificial, Yeast; D002896:Chromosomes, Human, Pair 6; D004247:DNA / Q000032:analysis; D015342:DNA Probes; D047108:Embryonic Development; D005260:Female; D005307:Fertilization in Vitro; D006579:Heterozygote; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007399:Interphase; D011247:Pregnancy
chemical_list: D015342:DNA Probes; D004247:DNA