Amniotic fluid alpha-fetoprotein as a marker in prenatal diagnosis of neural tube defects.

The prenatal diagnosis of anencephaly and spina bifida (neural tube defect, NTD) through amniotic fluid analysis for alpha-fetoprotein (AFP) is gradually gaining clinical recognition. AFP concentrations were determined in 237 amniotic fluids from normal pregnancies ranging between 7 and 42 weeks of gestation. A steady decline in AFP from 26 mug/ml at 7-9 weeks to 155 ng/ml at term is observed. AFP concentration was determined in 35 amniotic fluids from 33 confirmed neural tube defective pregnancies. In 14 cases where amniotic fluid was examined prior to the 26th week of gestation. AFP was markedly elevated when compared with the normal range of the same gestational period. In 21 amniotic fluids past the 26th week, 17 cases (85-) had markedly elevated AFP levels; however, 2 cases of anencephaly, 1 of spina bifida, and 1 of hydrocephaly gave levels within the normal range. It is concluded that elevated AFP in the amniotic fluid is a reliable but nonspecific marker for open neural tube defects prior to the 26th week of pregnancy, but may become normal after the 26th week in a small percentage of patients.     

Amniotic fluid fibrinogen degradation products in the prenatal diagnosis of neural tube defects.

Elevated levels of fibrin(ogen) degradation products (FDP) have been detected in second-trimester amniotic fluid in association with open neural tube defects (NTD'S) in the fetus. The FDP assay has been proposed for the prenatal diagnosis of NTD's and in this paper is compared with the established use of the alpha-fetoprotein (AFP) assay. FDP and AFP levels were measured in 132 amniotic fluid samples, including 20 NTD cases. The false negative and false positive rates for the FDP assay are high compared with the AFP assay, but it may be of value as an adjunct assay to reduce the over-all false positive rate.     

Amniotic fluid alpha-fetoprotein levels and prenatal diagnosis of autosomal trisomies

Pregnancies with fetal trisomy 21 have been associated with low amniotic fluid alpha-fetoprotein levels (AFAFP). This observation led to the suggestion that low AFAFP levels be used as a criterion for completion of a chromosomal analysis in patients who are not otherwise at increased risk for a fetal chromosome abnormality and in whom karyotyping might not have been completed for economic reasons. In order to assess the usefulness of such criteria, we reviewed the AFAFP levels of 90 cases of fetal trisomy 21, 23 cases of trisomy 18, and 10 cases of trisomy 13. These were compared with 2400 control samples with normal chromosome constitution. AFAFP levels were generally lower in pregnancies with trisomy 21, showing a median value of 0.72 MoM. However, 40 per cent of the trisomy 21 samples had AFAFP values greater than 0.8 MoM and 20 per cent were over 1.0 MoM. These data imply that over 50 per cent of Down syndrome cases might have been missed using a cut-off level of 0.70 MoM for completion of chromosome analysis. Using a higher cut-off level will leave only a small percentage of samples unkaryotyped. The distribution of AFP levels in trisomy 13 and 18 is no different from controls; we therefore believe that fetal karyotyping should be completed in every amniotic fluid sample obtained.     

Amniotic fluid acetylcholinesterase measurement in the prenatal diagnosis of open neural tube defects. Second report of the Collaborative Acetylcholinesterase Study

Seventeen centres from Australia, Britain, France, and the United States collaborated in a study to compare amniotic fluid acetylcholinesterase (AChE) determination by gel electrophoresis and amniotic fluid alpha-fetoprotein (AFP) measurement as diagnostic tests for open neural tube defects. The study was based on 32,642 women with singleton pregnancies (including 428 with open spina bifida and 238 with anencephaly) who had an amniocentesis at 13-24 weeks' gestation. The AChE test yielded a detection rate for open spina bifida of 99 per cent (95 per cent confidence interval 98-100 per cent), 98 per cent for anencephaly (95 per cent confidence interval 96-100 per cent), and a false-positive rate of 0.34 per cent (95 per cent confidence interval 0.28-0.40 per cent) excluding miscarriages, intrauterine death, and serious fetal abnormalities. The false-positive rate was 0.30 per cent among the 13 centres that used a specific AChE inhibitor in the test. Comparable rates for the AFP test were less favourable. (For example, the open spina bifida detection rate was 90 per cent and the false-positive rate was 0.46 per cent using the cut-off levels specified in the U.K. Collaborative AFP Study.) The AChE false-positive rate was lower in samples that were not bloodstained (0.16 per cent) than in those that were (2.4 per cent). It was higher in women who had an amniocentesis on account of a raised maternal serum AFP level (0.56 per cent) than in those who had one for other reasons (0.29 per cent). The best results were obtained by a combination of the two tests, an effective and economical policy being to perform the AFP measurement on all amniotic fluid samples and an AChE test on samples with AFP levels greater than or equal to 2.0 multiples of the normal median (about 5 per cent of all samples). Using this policy, the open spina bifida detection rate was 96 per cent and the false-positive rate was 0.14 per cent (0.06 per cent for samples that were not bloodstained and 1.2 per cent for those that were; 0.40 per cent for women with raised serum AFP levels and 0.09 per cent for other women). This policy offers a useful improvement to the prenatal diagnosis of open spina bifida.     

Amniography in the prenatal diagnosis of neural tube defects.

Amniography for the visualization and confirmation of suspected neural tube defect was performed in 9 midtrimester gravidas. In all cases, amniotic fluid alpha-feto protein (AFP) was abnormally elevated. Four cases of anencephaly and one of spina bifida were demonstrated by amniography. These pregnancies were terminated and the defects were confirmed by gross pathologic examination. In 4 remaining cases, amniography was normal. Three of these pregnancies proceeded to term, culminating in the birth of a normal child. The fourth patient had spontaneous abortion of a normal fetus at 23 weeks of gestation. The experience reported here suggests that amniography is an important adjunctive diagnostic technique in the prenatal diagnosis of neural tube defect, and if used correctly, may significantly reduce the chance of false-positive diagnosis.     

Amniotic fluid fibrinolytic system in fetal neural tube defects

Second trimester amniotic fluid fibrinolytic system was examined in normal pregnancies and those complicated by anencephaly, spina bifida and fetal chromosome abnormalities. No significant difference was demonstrated between the fibrinolytic systems from normal pregnancies and those complicated by fetal chromosome abnormalities. In pregnancies complicated with anencephaly and spina bifida no significant difference was demonstrated for alpha-1-antitrypsin, alpha-1-antichymotrypsin and urokinase. Plasminogen was significantly lower (p less than 0.02) and plasmin significantly higher (p less than 0.001) than levels from normal amniotic fluid. Alpha-2-macroglobulin, fibrinogen, FDP-D and FDP-E were detected only in pregnancies complicated with anencephaly and spina bifida.     

Maternal serum alpha-fetoprotein screening for open neural tube defects in twin pregnancies.

Data on maternal serum alpha-fetoprotein (AFP) levels at 13-24 weeks' gestation in 46 twin pregnancies with open neural tube defects (22 with anencephaly, 24 with open spina bifida) and 169 unaffected twins were used to estimate the detection and false-positive rates associated with different cut-off levels. Using the conventional cut-off level of 2.5 multiples of the median (MoM) for unaffected singleton pregnancies of the same gestation and laboratory, the detection rate in twins was 99 per cent for anencephaly and 89 per cent for open spina bifida, with a false-positive rate of 30 per cent. Using a 5.0 MoM cut-off level to maintain a similar false-positive rate to that found among singleton pregnancies at 16-18 weeks' gestation (about 3 per cent), the detection rate was 83 per cent for anencephaly and 39 per cent for open spina bifida. Estimates are provided of the odds of having an affected twin pregnancy given a positive AFP result as well as the odds for individual women with a raised AFP level.     

Amniotic fluid human chorionic gonadotrophin and alpha-fetoprotein levels in pregnancies conceived after assisted reproduction

OBJECTIVES: To study the alteration in the second-trimester maternal serum levels of human chorionic gonadotrophin (hCG) and alpha-fetoprotein (AFP) in pregnancies conceived after assisted reproduction. METHODS: We compared the amniotic fluid hCG and AFP concentrations of 45 pregnancies with fresh embryo transfer and 25 pregnancies with frozen-thawed embryo transfer with 269 spontaneous pregnancies. Wilcoxon rank-sum test was used for analysis. RESULTS: The median amniotic fluid hCG MoM in pregnancies conceived after frozen-thawed embryo transfer was significantly increased to 1.41 compared to 1.00 (p = 0.01) in naturally occurring pregnancies and 0.96 (p = 0.049) in pregnancies after fresh embryo transfer. Further analysis showed that this was only observed in frozen embryos fertilized by conventional insemination with MoM of 1.59. The AFP MoMs were similar among the groups. CONCLUSIONS: The observed raised amniotic fluid hCG level in IVF-FET pregnancies may reflect the elevated maternal serum level in these pregnancies. Further studies should be directed towards exploring the underlying pathophysiology.     

Amniotic fluid B12, calcium, and lead levels associated with neural tube defects

The group mean differences and relationships between folate, B12, calcium, lead, and methionine levels in amniotic fluid from 29 non-NTD and 11 NTD pregnancies (15-20 weeks' gestation age) were determined. The study population was predominantly Hispanic in both groups (48% in non-NTD and 36% in NTD group) with an average maternal age of 29 years in the non-NTD group and 24 years in the NTD group; and, an average gestation age of 18.5 weeks in the non-NTD and 17.5 weeks in the NTD group. The folate, B12, lead, calcium, and methionine levels of the two groups were compared by Student's t-test and by Pearson's correlation. The NTD levels were lower for calcium (p< or =0.0001), B12 (p< or =0.001), and methionine (P< or =0.001); but, the lead level was higher (p< or =0.0001). A negative correlation was observed between lead and both B12 (p< or =0.007) and methionine (p< or =0.02).     

Prenatal diagnosis of neural tube defects. IV. Maternal serum alpha-fetoprotein screening

The maternal serum alpha-fetoprotein (AFP) in 6161 women in routine pregnancy [2771 in a hospital obstetric clinic (group 1) and 3390 in private practices (group 2)] was studied. Group 1 studies enabled the delineation of the normal range of serum AFP, whereas group 2 represented a true screening experience. In group 2, 39 (2.5%) of 1566 women at 16 to 18 weeks' gestation had raised (2.5 times the median or more) serum AFP. Of these 39 women, 3 (7.8%) had neural tube defects (NTDs), 6 (15.4%) had multiple pregnancies, 1 (2.6%) had congenital nephrosis, 7 (17.9%) had spontaneous abortions, 7 (17.9%) had miscellaneous associated factors, and 15 (38.5%) had raised serum AFP for no obvious reason. Only 16 (1%) women had "unnecessary" amniocenteses. None of these aborted subsequently. Analysis of the combined data showed that NTDs were detectable in 87.5% of patients-all 6 with anencephaly and 1 of 2 with spina bifida (1 spina bifida lesion closed); multiple pregnancy was determined in 45% (18/40 cases), and spontaneous abortion ensued in 14.5%. In group 1 a raised serum AFP was associated with a host of complications in 77.3% of the women. Low AFP values had associated complications in 72.2% of cases. Maternal serum AFP screening represents another potentially important tool for early detection of high-risk pregnancy.     

Amniotic fluid alpha-fetoprotein and acetylcholinesterase measurements in pregnancies associated with gastroschisis

In this study we define for the first time the distribution of alpha-fetoprotein levels and acetylcholinesterase ratios in amniotic fluid samples associated with fetal gastroschisis. Elevated alpha-fetoprotein levels and positive acetylcholinesterase measurements are found in virtually all cases, indicating that these combined measurements are highly reliable in detecting this lesion.     

Maternal serum alpha-fetoprotein and previous neural tube defects.

A case-control study involving 63 pregnancies showed that women who had previously had an infant with anencephaly or spina bifida did not have raised serum alpha-fetroprotein (AFP) levels in subsequent unaffected pregnancies. The value of a serum AFP determination in such women is discussed. If the serum AFP level is normal and if ultrasonography excludes anencephaly the risk of spina bifida might be low enough (about 1 per cent) to make diagnostic amniocentesis difficult to justify. Conversely, a high serum AFP value in such women should not be acted on without confirmation of an abnormality by ultrasonography and, if that is negative, by diagnostic amniocentesis.     

Prenatal diagnosis of neural tube defects. III. A reevaluation of the alpha-fetoprotein assay.

Alpha-fetoprotein was measured in 2209 amniotic fluid samples under improved assay conditions. All cases of open neural tube defects had elevated values greater than 5 SD above the mean. The false positive rate based on the 3 SD cutoff is estimated at less than 9.15%. Alpha-fetoprotein assays are recommended for all patients undergoing second trimester amniocentesis, optimally between 14 and 16 weeks' gestation.     

Amniotic fluid levels of fibrin(ogen) degradation fragment E and alpha-fetoprotein in normal pregnancy and with fetal neural tube defect.

Amniotic fluid levels of alpha-fetoprotein (AFP) and fibrin(ogen) degradation fragment E (FgE) were measured in 214 normal subjects and 27 pregnancies associated with an abnormal fetus (open neural tube defect or exomphalos). AFP levels showed no overlap between the normal and abnormal groups, thus confirming the reliability of the AFP assay in the detection of these abnormalities. FgE levels however showed considerable overlap and could not be used as a diagnostic parameter. Agarose gel chromatography of the amniotic fluid revealed the presence of large molecular weight FgE related antigen in open neural tube defect suggesting that leakage of proteins from exposed capillaries is responsible for the elevation of amniotic fluid protein levels.     

Maternal serum vitamin D levels in pregnancies complicated by neural tube defects

Objective: The association between vitamin D deficiency and abnormal neural development has been proposed previously. We aimed to evaluate maternal serum vitamin D levels in pregnancies complicated by neural tube defects (NTDs) and compared them with healthy pregnant women.

Methods: A total of 60 pregnant women were included in this controlled cross-sectional study. Thirty of the patients whose pregnancies were complicated by meningocele, meningomyelocele, encephalocele, anencephaly and fetal acrania constituted the study group, whereas 30 normal pregnant women constituted the control group. The main parameters recorded for each woman were as follows: age, body mass index (BMI), gestational week (GW), gravidity, abortion, co-morbidities, dressing style, consumption of milk and dairy products and serum levels of 25(OH)VitD₃, calcium, albumin and total protein.

Results: The mean maternal serum 25(OH)VitD₃ level was 6.2?±?5.0?ng/ml in the study group and 9.1?±?7.3?ng/ml in the control group (p: 0.071). The mean maternal serum calcium level was statistically significantly higher in the control group, and calcium-rich dietary intake was also more common in this group (p?<?0.05). There was no statistically significant difference between groups in terms of age, BMI, GW, dressing style and serum levels of albumin and total protein.

Conclusions: Vitamin D deficiency is common among pregnant women, and maternal serum calcium levels were lower in pregnancies complicated by NTD than healthy pregnant women. Larger further studies are required to evaluate the effects of calcium-rich dietary sources or vitamin D and calcium in the development of NTDs.