Because there is no reliable method except elective abortion in treating pregnant women who have fetus(es) with severely compromised genetic disease(s), and the prognosis of the fetus depends upon sex, the determiniation of fetal sex as early in
gestation as possible is most important.
It is well known that polymerase chain reaction is a useful method of prenatal genetic diagnosis in molecular levels. Specific amplification ZFY region which is male specific within the short arm of Y chromsome was performed to determine the
fetal
sex
rapidly and accurately.
In this study, a buffer containing 2.5mM MgCl2 , 100μM dNTPs, and annealing temperature 44℃were found to yield the maximum product, and a successful amplification was achieved with only 1 cc of amniotic fluid sample. Six male DNA from
peripheral
blood
generated a 530bp band and 4 female DNA produced no band. Among the 10 amniotic fluid samples, 8 generated a 530bp band and remaining 2 samples produced no band. The results of chromosome analysis on amniotic cell cultures confirmed that all
samples
having the 530bp band had a 46, XY karyotype whereas the samples showing no band were 46 XX. Among the 5 chorionic-villus samples, 2 generated q 530bp band and remaining 3 samples produced no band. The results of chromosome analysis on chorionic
villus
cultures confirmed that 2 samples having the 530bp band had a 46, XY karyotype whereas 2 of 3 samples showing no band were 46,XX and remaining one was 46, XX,t(14:21). Successful amplification was obtained with simple lysing method instead of
complicated DNA purification.
These data suggest that PCR amplification of ZFY region can make a rapid and correct fetal sex determination, and may be used for basic tools of prenatal genetic diagnosis.
|