Rapid sexing of human embryos by non‐radioactive in situ hybridization: Potential for preimplantation diagnosis of X‐linked disorders

Abstract

Sixty spare human embryos at various stages of preimplantation development were prepared for cytogenetic analysis. Fluorescent staining of those with metaphases allowed scoring for the presence of a Y chromosome. In situ hybridization was then performed using a biotinylated Y-specific sequence, and the probe was detected by a standard streptavidin-linked alkaline phosphatase system. This enabled comparison of the chromosomal sex with that obtained after in situ hybridization in 28 embryos, and the sexing result obtained by the two methods was concordant in all cases. A further 21 embryos in which no metaphase chromosomes were obtained were sexed by biotinylated in situ hybridization only. Overall, 66 per cent of male interphase nuclei demonstrated a Y-specific hybridization signal. Results were obtained in under 24 h, which may permit the sexing of an embryo biopsied during cleavage and the transfer of sexed embryos at the blastocyst stage to the mother's uterus in the same cycle as oocytes are collected for in vitro fertilization.

Rapid Prenatal Diagnosis of Chromosomal Aneuploidies by Fluorescence in Situ Hybridization: Clinical Experience With 4500 Specimens

“…Procedures which allow rapid detection of certain chromosomal abnormalities in uncultured fetal cells have been known for some time, as demonstrated by analysis of Barr and Y bodies to detect sex chromosome aneuploidies (Robinson et al 1979;Penketh et al 1989;West et al 1989). Fluorescence in situ hybridization (FISH) to interphase nuclei with chromosome-specific DNA probes can now rapidly and accurately detect the most common autosomal trisomies and aneuploidies of the sex chromosomes (Cremer et al 1986;Lichter et al 1988aLichter et al , 1988bKlinger et al 1990Klinger et al , 1992 Christensen et al 1992; Lebo et al 1992;Zahed et al 1992;Zheng et al 1992).…”

Multiplex Polymerase Chain Reaction Used for Bovine Embryo Sex Determination

“…Dev. 57: [539][540][541][542] 2011) ex determination of preimplantation embryos is important for manipulating the sex ratios of domestic animals at an early stage of embryo development [1,2]. A number of approaches have been used to determine the sex of bovine embryos but have failed to gain much popularity due to the lack of either accuracy or speed [3].…”

Pre-Implantation Development

“…Initially, the procedure utilized radiolabelled probes coupled with autoradiography, and was used to detect Y chromosome-specific repeated nucleotide sequences in the human male embryo Oones et al, 1987) and in domestic species (Leonard et al, 1987;Matthews et al, 1987). More recently, rapid non-isotopic methods have been used for the detection of both X-and Y -chromosome sequences in human preimplantation embryos (Penketh et al, 1989;Criffin et al, 1992). Since signals are visualized directly under the microsope, localization of the signal is more precise.…”