An accurate and rapid gender determination assay in single cells by the capillary polymerase chain reaction method.
OBJECTIVE
In preimplantation genetic diagnosis (PGD), a rapid and accurate assay has been required. We have therefore developed a capillary polymerase chain reaction (PCR) method using rapid thermal cycling programs to determine the gender of single amniocytes.


METHODS
Single amniocytes from each amniotic fluid sample were isolated by micromanipulation and their gender was determined by a multiplex PCR assay in a capillary tube, using primers that amplify a 308-bp DXZ1 and a 154-bp DYZ1 repeat sequence on the X and Y chromosomes, respectively.


RESULTS
All four thermal cycling programs, which took 180, 150, 120, and 90 min, were 100% accurate in diagnosing the gender of single amniocytes. No DNA contamination was observed in any samples.


CONCLUSIONS
The multiplex PCR assay was rapid and accurate in diagnosing gender in single cells and may be clinically applicable in PGD.
Hashiba
T
T
Department of Obstetrics and Gynecology, Keio University School of Medicine, Tokyo, Japan.
Sueoka
K
K
Kuroshima
M
M
Asada
H
H
Kuji
N
N
Yoshimura
Y
Y
D000650:Amnion / Q000166:cytology*; D005260:Female; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D019836:Preimplantation Diagnosis / Q000379:methods; D019849:Sex Determination Processes