Polymerase-chain-reaction-based detection of fetal rhesus D and Y-chromosome-specific DNA in the whole blood of pregnant women during different trimesters of pregnancy.
OBJECTIVE
The aim of this study was to determine whether or not a noninvasive procedure utilizing maternal peripheral blood as the source of DNA and polymerase chain reaction (PCR) could be used to detect fetal rhesus D (RhD) status as well as fetal gender during different gestational stages of pregnancy.


METHODS
Maternal blood samples were obtained from 54 RhD-negative pregnant women during the first trimester (6-13 weeks, n = 14), second trimester (14-26 weeks, n = 26) and third trimester (27-40 weeks, n = 14). Genomic DNA was extracted from the whole blood and analyzed by seminested and nested PCR for detection of DNA sequences corresponding to RhD (n = 54) and Y chromosome (n = 48) using RhD and Y-chromosome-specific oligonucleotide primers, respectively. The seminested/nested PCR results were compared with the RhD status and gender of the babies after delivery.


RESULTS
The sensitivity and specificity of seminested PCR for detection of fetal RhD positivity in whole blood of pregnant women were 81 and 100%, respectively, while the sensitivity and specificity of nested PCR for detection of male fetuses, using Y-chromosome-specific DNA as a marker, were 96 and 91%, respectively. There were no significant differences in the PCR results with samples obtained from women at different gestational stages of pregnancy.


CONCLUSIONS
Seminested and nested PCRs for detection of fetal RhD and gender status, respectively, by using the blood of pregnant women during different gestational stages of pregnancy, are reliable noninvasive procedures with high sensitivity and specificity.
Al-Yatama
M K
MK
Department of Obstetrics and Gynecology, Faculty of Medicine, Kuwait University, Kuwait.
Mustafa
A S
AS
Al-Kandari
F M
FM
Khaja
N
N
Zohra
K
K
Monem
R A
RA
Abraham
S
S
D041322:Chromosomes, Human, Y; D004247:DNA / Q000097:blood*; D005260:Female; D005312:Fetal Blood / Q000737:chemistry*; D005865:Gestational Age; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D011247:Pregnancy; D011264:Pregnancy Trimesters / Q000097:blood*; D011296:Prenatal Diagnosis / Q000379:methods*; D012203:Rh Isoimmunization / Q000097:blood / Q000235:genetics; D012204:Rh-Hr Blood-Group System / Q000097:blood* / Q000235:genetics; D012680:Sensitivity and Specificity; D012732:Sex Determination Analysis / Q000379:methods
D012204:Rh-Hr Blood-Group System; C026410:Rho(D) antigen; D004247:DNA