[Usefulness of quantitative assessment of Toxoplasma gondii genome using PCR-TaqMan in amniotic fluid, maternal and neonatal blood in selected complications in pregnancy].
Routine serological diagnosis of Toxoplasmosis provides high sensitivity, but not high specificity. The high sensitivity combined with high specificity offered by PCR-TaqMan as well as the degree of infection led us to investigate the presence and levels of Toxoplasma gondii genome in amniotic fluid, maternal and neonatal blood in cases of pregnancy where infection with this agent was suspected.


METHODS
Samples of amniotic fluid and blood were taken from 28 women between the 16th and 40th week of gestational age. Postnatal blood samples were also taken from their infants. Included in the study group were women with IUGR, PROM, preterm delivery imminent, Toxoplasmosis in previous pregnancies, raised IgG or IgM anti-Toxoplasma antibody titers and with amniotic fluid disturbances (oligohydramnios and polyhydramnios). Presence and levels of Toxoplasma genome was investigated using PCR-TaqMan. PCR products were detected by electrophoresis on polyacrylamide gel.


RESULTS
Toxoplasma gondii genome was detected in blood from 13 women, 3 newborns and in amniotic fluid from one other women. Toxoplasma genome was detected in blood from one newborn, but was not detected in sample from its mother.


CONCLUSIONS
The PCR-TaqMan test is highly sensitive and specific method allowing detection of the parasite genome and assessment of its level. Limitations of this method are its relatively high cost and poor access to ABI PRISM 7700 (TaqMan) sequence detector. The PCR TaqMan is useful in cases, where serological tests for the presence of infections are ambiguous.
Gola
J
J
Katedra i Klinika Patologii Ciazy i Rozrodczości w Zabrzu.
Czuba
B
B
Mazurek
U
U
Sławska
H
H
Kamiński
K
K
Wilczok
T
T
D000653:Amniotic Fluid / Q000469:parasitology*; D000818:Animals; D005260:Female; D005317:Fetal Growth Retardation / Q000097:blood; D005322:Premature Rupture of Fetal Membranes / Q000097:blood; D018503:Genome, Protozoan; D006801:Humans; D007231:Infant, Newborn; D016133:Polymerase Chain Reaction / Q000379:methods; D011247:Pregnancy; D015597:Pregnancy Complications, Parasitic / Q000097:blood / Q000175:diagnosis / Q000469:parasitology; D012680:Sensitivity and Specificity; D019914:Taq Polymerase / Q000235:genetics; D014122:Toxoplasma / Q000235:genetics*; D014125:Toxoplasmosis, Congenital / Q000097:blood / Q000175:diagnosis / Q000469:parasitology
D019914:Taq Polymerase