Sex determination by simultaneous application of polymerase chain reaction and fluorescent in situ hybridization on the same blastomere of a pre-embryo.
OBJECTIVE
To describe an investigation of the feasibility of carrying out polymerase chain reaction (PCR) and fluorescent in situ hybridization on the same single human cell for sex determination.


METHODS
To develop protocol for a clinical diagnostic test in preclinical trials.


METHODS
Infertile human volunteers in a clinical environment.


METHODS
Polyploid embryos were obtained from patients treated by IVF at the Advanced Institute of Fertility.


METHODS
Seventeen blastomeres biopsied from human polyploid embryos were analyzed for sex determination by simultaneous application of PCR and fluorescent in situ hybridization.


METHODS
Presence of an X- or X and Y-chromosome band after agarose gel electrophoresis of PCR products and presence of an X- or X and Y-chromosome fluorescent probe signal after fluorescent in situ hybridization following PCR analysis.


RESULTS
By PCR, all 17 blastomeres were amplified and, by fluorescent in situ hybridization, 12 (70.6%) blastomeres produced signals that were consistent with PCR results. Two blastomeres showed only X signals, although by PCR they had both X and Y-chromosome bands.


CONCLUSIONS
The sequential use of PCR and fluorescent in situ hybridization on the same blastomere can be applied to improve the accuracy of sex determination before fresh ET.
Sasabe
Y
Y
Advanced Institute of Fertility, Arlington Heights, Illinois, USA.
Krisher
R L
RL
Stehlik
J C
JC
Stehlik
E F
EF
Katayama
K P
KP
D001483:Base Sequence; D001757:Blastomeres / Q000737:chemistry / Q000166:cytology*; D004247:DNA / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D004622:Embryo, Mammalian / Q000737:chemistry / Q000166:cytology*; D005260:Female; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012732:Sex Determination Analysis / Q000379:methods*; D014960:X Chromosome; D014998:Y Chromosome
D004247:DNA