Extraction of DNA from amniotic fluid cells for the early prenatal diagnosis of genetic disease.
Ten-ml samples of amniotic fluid were taken from pregnancies being terminated at 8-14 weeks' gestation. DNA was extracted from the amniotic cells by sequential centrifugation and analysed using the polymerase chain reaction (PCR). Fifteen samples were analysed for evidence of maternal contamination using Mfd5 oligonucleotide primers for repeat polymorphisms. Ten amniotic fluid samples were tested for the Delta-F508 deletion characteristic of cystic fibrosis to demonstrate a diagnostic application for the technique. In each case, DNA extracted from fetal tissue from the same pregnancy was included in the controls. In 14 of the 15 cases tested with the Mfd5 primers, both the amniotic fluid DNA and the fetal DNA showed no evidence of contaminating DNA. In one case, neither the amniotic fluid cells nor the fetal cells yielded results. In nine of the ten cases tested with the Delta-F508 primers, the amniotic fluid cell DNA provided accurate information about the genetic status of the fetus; in the tenth, the fetal DNA failed to amplify. The results indicate that adequate DNA can be extracted from amniotic fluid from 8 weeks' gestation onward and these samples are suitable for prenatal diagnosis using PCR.
Rebello
M T
MT
Department of Cytogenetics, St Mary's Hospital, Paddington, London, U.K.
Hackett
G
G
Smith
J
J
Loeffler
F E
FE
Robson
S
S
MacLachlan
N
N
Beard
R W
RW
Rodeck
C H
CH
Williamson
R
R
Coleman
D V
DV
D000328:Adult; D000653:Amniotic Fluid / Q000166:cytology*; D002869:Chromosome Aberrations / Q000175:diagnosis*; D025063:Chromosome Disorders; D004247:DNA / Q000302:isolation & purification*; D005260:Female; D006801:Humans; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D011296:Prenatal Diagnosis / Q000379:methods*
D004247:DNA