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Rapid sexing of human embryos by non-radioactive in situ hybridization: Potential for preimplantation diagnosis of X-linked disorders |
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| Authors: | R J A Penketh J D A Delhanty J A van den Berghe E M Finklestone A H Handyside S Malcolm R M L Winston |
| Institution: | 1. Institute of Obstetrics and Gynaecology, Hammersmith Hospital, Du Cane Road, London W12 OHS
Seconded from Clinical Research Centre, Watford Road, Harrow, Middlesex, U.K.;2. The Galton Laboratory, Department of Genetics and Biometry, University College London, 4 Stephenson Way, London NW1 2HE;3. Institute of Child Health, 30 Guilford Street, London WCIN IEH, U.K.;4. Institute of Obstetrics and Gynaecology, Hammersmith Hospital, Du Cane Road, London W12 OHS |
| 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 streptavidinlinked 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. |
| Keywords: | Preimplantation diagnosis Embryo sexing Non-radioactive in situ hybridization |