François Forestier

Prenatal Diagnosis of Congenital Toxoplasmosis with a Polymerase-Chain-Reaction Test on Amniotic Fluid

BACKGROUND Congenital infection with Toxoplasma gondii can produce serious sequelae. However, there is little consensus about screening during pregnancy, and the tests used to establish a prenatal diagnosis of toxoplasmosis are complex and slow. We evaluated a simpler approach that is based on a polymerase-chain-reaction (PCR) test. METHODS Prenatal diagnostic tests, including ultrasonography, amniocentesis, and fetal-blood sampling, were performed in 2632 women with T. gondii infection acquired during pregnancy. In 339 consecutive women, a competitive PCR test for T. gondii was performed on amniotic fluid, and its results were compared with those of conventional diagnostic tests. The PCR test targets the B1 gene of T. gondii, uses an internal control, and can be completed in a day. Positive tests were confirmed by serologic testing of newborns or by autopsy in terminated pregnancies. RESULTS Overall, the risk of fetal infection was 7.4 percent, but it increased sharply with gestational age. Congenital infection was demonstrated in 34 of 339 fetuses by conventional methods, and the PCR test was positive in all 34. In three other fetuses, only the PCR test gave positive results, and follow-up testing confirmed the presence of congenital toxoplasmosis. The PCR test gave one false negative result but no false positive results. The PCR test performed better than conventional parasitologic methods (sensitivity, 97.4 vs. 89.5 percent; negative predictive value, 99.7 vs. 98.7 percent). CONCLUSIONS For the prenatal diagnosis of congenital T. gondii infection, an approach based on a PCR test performed on amniotic fluid is rapid, safe, and accurate.

Prenatal management of 746 pregnancies at risk for congenital toxoplasmosis

Abstract When infection with Toxoplasma gondii occurs during pregnancy, there is a risk that the parasite will cause severe congenital toxoplasmosis. We developed a method of diagnosing and treating congenital toxoplasmosis in utero. Diagnosis was based on the identification of maternal acute infection, followed by culture of fetal blood and amniotic fluid, testing of fetal blood for toxoplasma-specific IgM and nonspecific measures of infection, and ultrasound examination of the fetal brain. Treatment included the administration of antibiotics to all mothers with confirmed acute infection during pregnancy, with more intensive antibiotic treatment of those who had infected fetuses and who chose to continue the pregnancy. We report a prospective study of 746 documented cases of maternal toxoplasma infection, in which the infants were followed for at least three months. Infection was diagnosed antenatally in 39 of 42 fetuses. Twenty-four of the 39 pregnancies were terminated, and 15 were continued. All the m...

Serum Insulin-Like Growth Factors and Insulin-Like Growth Factor Binding Proteins in the Human Fetus. Relationships with Growth in Normal Subjects and in Subjects with Intrauterine Growth Retardation

ABSTRACT: IGF-I, IGF-II, and their binding proteins (BP) were studied in sera obtained by direct puncture of umbilical cords in utero between 20 and 37 wk of gestation in 103 normal fetuses and in 16 fetuses with intrauterine growth retardation, as well as in the cord blood of 37 normal newborns of 38− to 42-wk pregnancies. In normal fetuses, IGF-I levels were approximately 50 ng/mL and IGF-II levels approximately 350 ng/mL up to the 33rd wk of pregnancy. Thereafter, both increased to reach values two to three times higher at term. Correlations were found between fetal placental lactogen levels and those of IGF-I and IGF-II, which is consistent with the hypothesis that placental lactogen is involved in the regulation of IGF synthesis in the fetus. With weight (either measured at birth or deduced from echographical data) as index of fetal size, IGF-I levels were significantly (p < 0.001) higher in fetuses with weights above the mean for gestational age than in fetuses with weights below the mean, whereas IGF-II levels were similar in the two groups. Similarly, IGF-I (but not IGF-II) levels in fetuses with intrauterine growth retardation were significantly lower than those in normal fetuses of the same age (p < 0.01). These findings suggest that, during the latter months of intrauterine life, IGF-I (but not IGF-II) is involved in the control of fetal size. Total fetal BP concentrations were approximately 1/3 those of adults. The fetal electrophoretic profile obtained by Western-ligand blotting bore a strong resemblance to that of subjects with growth hormone deficiency. In new borns, the proportions of IGF-I and IGF-II associated with BP to form 150-kD complexes were considerably lower than those in adults, but similar to those in hypopituitary patients. It may be deduced from these findings that during fetal life, BP synthesis is adapted to increase the bioavailability of the IGF at a time when growth is at a maximum.

Fetal toxoplasmosis: outcome of pregnancy and infant follow-up after in utero treatment.

Eighty-nine cases of fetal Toxoplasma infection are reported in women treated with spiramycin during pregnancy. Thirty-four pregnancy terminations were performed (2.7% of the total number of acquired Toxoplasma infections during pregnancy). Fifty-two pregnancies were allowed to proceed (43 being additionally treated with pyrimethamine and sulfonamides), leading to the birth of 54 live infants. After a mean follow-up period of 19 months, 41 infants had evidence of subclinical Toxoplasma infection, 12 had a benign form, and one had severe congenital toxoplasmosis (this infant did not receive the additional treatment during pregnancy). Efficacy of the additional treatment with pyrimethamine and sulfonamides was demonstrated by a significant reduction of severe congenital toxoplasmosis and the relative decrease of the ratio of benign to subclinical forms. We recommend that spiramycin treatment be started as soon as possible once the diagnosis of maternal Toxoplasma infection during pregnancy is proved or strongly suspected, because a prolonged time interval between onset of infection and start of treatment seems to be associated with the presence of severe fetal lesions at the time of prenatal diagnosis.

Hematological Values of 163 Normal Fetuses between 18 and 30 Weeks of Gestation

ABSTRACT. Utilizing an easy and safe procedure for fetal blood sampling in utero we have studied 409 fetuses for prenatal diagnosis of rubella, toxoplasmosis, hemophilia, and hemoglobinopathies. Retrospectively we selected 163 fetuses confirmed as normal at birth and tested between 18 and 30 wk of gestation to establish normal hematological parameters and to follow the evolution of erythropoiesis, differential counts, hemoglobin synthesis, and hemostasis. Total white blood cell and platelet counts did not change during this period. The lymphocytes represented the main population and we observed a decrease of normoblasts during gestation. The results show a progressive increase of red blood cells and hemoglobin. This evolution is demonstrated by the ratio hemoglobin A to acetylated hemoglobin F. No significant modification of hemostasis was observed over a 12-wk intrauterine gestation. These results provide useful reference values for future investigations.

Fetal platelet counts in thrombocytopenic pregnancy

Fetal platelet counts were assessed by percutaneous umbilical blood sampling in 64 pregnancies (62 women) with maternal thrombocytopenia. In 33 pregnancies associated with chronic immune thrombocytopenia, 11 of the fetuses had platelet counts below 150 x 10(9)/l and 4 were severely thrombocytopenic (less than 50 x 10(9)/l). In 31 pregnancies with symptomless maternal thrombocytopenia as an incidental finding, 4 fetuses were thrombocytopenic, 1 of them severely. Maternal indices, including antiplatelet antibodies, did not correlate with risk of fetal thrombocytopenia; and in those with repeat measurements there was no evidence of benefit from treatment with either corticosteroids (4 cases) or intravenous immunoglobulin (3 cases). Percutaneous umbilical blood sampling, a safe procedure in experienced hands, provides accurate platelet counts in thrombocytopenic pregnancy, as an aid to decisions on mode of delivery and to assessment of treatments.

Prenatal diagnosis and management of bleeding disorders with fetal blood sampling

The technique of fetal blood sampling for prenatal diagnosis has been shown to be both feasible and safe. The availability of fetal blood for direct evaluation has changed our attitude about the problems of both hereditary and acquired immune fetal bleeding disorders. We can continue with the classic approach and use fetal blood sampling for those conditions in which termination may be recommended, but we can also investigate less severe disorders in which the diagnosis allows us to plan the management of the pregnancy and minimize intrapartum and neonatal complications. We report our experience in prenatal diagnosis and management of 103 cases of hereditary and 18 cases of acquired immune bleeding disorders. We have developed specific management plans depending on the disorder under investigation, the severity of the condition in the fetus, and parental wishes. We have performed in utero transfusions of platelets and factor concentrate where appropriate. Efficacy of maternal therapy for fetal conditions can be directly assessed during gestation. Mode of delivery is determined by obstetric conditions and fetal status, directly assessed after appropriate therapy. Closer surveillance of the fetus by fetal blood sampling gives precise information on which to base clinical decisions to provide optimal maternal and fetal outcome.

PRENATAL TREATMENT OF ALLOIMMUNE THROMBOCYTOPENIA

absorption experiments, and there was no relation with previous hormonal treatment. Several disorders are associated with circulating autoantibodies. These autoantibodies often react with cells or tissues from healthy donors, but, except in a few cases, the antigen determinant has not been characterised. The significance of our finding remains to be worked out. The cause of undescended testis is still unclear in cases where mechanical factors are absent,4 but there may be a disturbance of the hypothalamo-pituitary testicular axis with an early deficiency of LH secretion.5 Testicular descent usually happens in the last week in utero, so if autoantibodies play a role in maldescent they would have to act on the fetus. Preliminary results suggest that this may be so. We have detected such antibodies in 5 out of 12 mothers of cryptorchid children tested during the first week after delivery. The same antibodies ofIgG class were detected in the newborn babies. Longitudinal and collaborative studies are in

The assessment of fetal blood samples

Fetal blood sampling under ultrasound control is rapidly expanding the study of human fetal biology. Pure fetal blood is required for prenatal diagnosis, establishment of reference ranges for biologic measurements, and assessment of fetal welfare. We present here the methods that we have developed to detect contamination in more than 1500 samples. These include hematologic indexes, blood smear, erythrocyte antigens, beta-human chorionic gonadotropin, and coagulation factor assays. The tests are relatively simple, inexpensive, and widely available. No single test is reliable in all situations, and it is necessary to perform all the tests on each sample of fetal blood. In the clinical setting where irrevocable action may be taken as a consequence of our results, we require absolute assurance of the purity of the sample.

Blood Chemistry of Normal Human Fetuses at Midtrimester of Pregnancy

Abstract: Thirteen biochemical parameters and five enzymatic activities were determined on sera of 63 normal human fetuses sampled by direct puncture under ultrasound guidance, between the 20th and the 26th wk of gestation, and on their mothers. They were referred to us for various prenatal diagnoses but were well and confirmed healthy at birth. Some parameters were found to be very similar in both groups, mainly creatinine, calcium, creatine kinase, aspartate aminotransferase, and γ-glutamyl transferase. Some values were significantly higher in the fetuses, such as total bilirubin, direct bilirubin, phosphorus, lactic dehydrogenase and alkaline phosphatase activities, and α-fetoprotein. Urea, uric acid, glucose, triglycerides, cholesterol, total protein, and albumin levels were found to be lower in fetuses. These data indicate a slower metabolism in fetuses compared to their mothers, a lower level of energy requirement, and a relative liver immaturity. These normal values of fetal biochemistry will improve our knowledge of physiology and help to determine the specific values of a test in fetal pathology.