Reliability of gender determination using the polymerase chain reaction (PCR) for single cells.
Contamination with extraneous DNA sequences is a frequent problem when performing PCR analysis of single cells. This report describes our experience with eliminating contaminating DNA sequences from PCR reagents for the purposes of gender identification. We have used amplification of Y-specific sequences to identify the gender of single human amniocytes. Female cells consistently showed no Y-specific bands but only 80% of male cells showed the expected intense Y-specific band. This phenomenon could lead to incorrect gender identification of single cells. We developed a technique of simultaneous amplification of X- and Y-specific sequences to prevent misdiagnosis because of failed PCR, which allows accurate preimplantation gender determination for women at risk for conceiving children with X-linked genetic diseases. We analyzed the gender of 141 consecutive single cells in a blinded manner without a single incorrect gender assignment.
Strom
C M
CM
Reproductive Genetics Institute, Illinois Masonic Medical Center, Chicago 60657.
Rechitsky
S
S
Verlinsky
Y
Y
D001757:Blastomeres; D015342:DNA Probes; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D011296:Prenatal Diagnosis; D015203:Reproducibility of Results; D012732:Sex Determination Analysis
D015342:DNA Probes