High sensitivity of fetal DNA in plasma compared to serum and nucleated cells using unnested PCR in maternal blood.
DNA analysis of blood is conventionally performed on cells - plasma and serum are discarded. Free DNA has been demonstrated in serum in cancer and autoimmune disorders and in pregnancy. We investigated possible noninvasive prenatal diagnosis using fetal DNA from maternal plasma and serum in pregnancy. Fetal gender was determined by PCR on DNA from maternal venous blood, serum and plasma of 65 women by boiling with or without phenol/chloroform extraction. When sensitivities were compared for plasma, additional phenol/chloroform extraction proved more sensitive than boiling alone (89 vs. 50%), the observed difference was 50% (CI 19 to 81%). Extracted plasma amplified better than extracted serum (89 vs. 46%), the observed difference being 44% (CI 22 to 66%). In contrast, fetal gender could not reliably be determined from DNA extracted from maternal nucleated blood cells. The size of plasma and serum DNA at 15-17 weeks of gestation was >1,500 bp. This work confirms the presence of fetal DNA in maternal plasma and serum which may be applicable to noninvasive prenatal diagnosis of paternally derived DNA sequences. We conclude that optimal sensitivity requires two methods of DNA extraction and that the use of plasma is preferred to that of serum.
Houfflin-Debarge
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V
Institute of Obstetrics and Gynaecology, Department of Materno-Fetal Medicine, Imperial College School of Medicine, Queen Charlotte's and Chelsea Hospital, London, UK.
O'Donnell
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H
Overton
T
T
Bennett
P R
PR
Fisk
N M
NM
D002725:Chloroform; D004247:DNA / Q000097:blood*; D005260:Female; D005333:Fetus; D005865:Gestational Age; D006358:Hot Temperature; D006801:Humans; D019800:Phenol; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011296:Prenatal Diagnosis; D012680:Sensitivity and Specificity; D012732:Sex Determination Analysis
D019800:Phenol; D002725:Chloroform; D004247:DNA