Clinical Trial Details
— Status: Completed
Administrative data
| NCT number |
NCT01232387 |
| Other study ID # |
10-0920 |
| Secondary ID |
KL2RR029885 |
| Status |
Completed |
| Phase |
N/A
|
| First received |
October 29, 2010 |
| Last updated |
September 19, 2013 |
| Start date |
May 2011 |
| Est. completion date |
September 2013 |
Study information
| Verified date |
September 2013 |
| Source |
Icahn School of Medicine at Mount Sinai |
| Contact |
n/a |
| Is FDA regulated |
No |
| Health authority |
United States: Institutional Review Board |
| Study type |
Observational
|
Clinical Trial Summary
Objectives: 1) To determine risk factors for fetomaternal hemorrhage. 2) To identify a
cost-effective method to detect fetomaternal hemorrhage prior to significant fetal anemia.
Significance/Background: Fetomaternal hemorrhage (FMH) is a condition in which occurs when
the placenta transfers blood from the fetus to the mother. Normally, nutrition and gasses
pass from mother to baby through the placenta and only waste products pass from baby to
mother through the placenta. Whole blood cells do not normally cross the placenta in
significant amounts. Mild FMH, where a small amount of whole blood passes from fetus to
mother but does not hurt the mother or baby, occurs in about 75% of pregnancies. A pregnant
woman does not know this occurs. It is only discovered if a special blood test that is
labor-intensive to perform and difficult to interpret called the Kleihauer-Betke acid
elution test is done. As mild FMH hurts no one, this test is not part of routine care. In
most cases, testing is done only if a baby is born sick with unexplained anemia. Severe FMH,
which can cause the baby to become sick from anemia (low red blood cell count) is caused by
large blood loss into the mother, occurs in only 1-3 per 1000 births. Severe anemia caused
by FMH can result in death of the baby before or after birth, or significant illness in the
newborn period. Short term problems for the baby include difficulty breathing, difficulty
maintaining blood pressure, and difficulty providing oxygen to all parts of the body. This
can cause multiple problems with the function of internal organs including the liver,
kidneys, intestines, and brain. Babies who become sick from severe FMH can develop long-term
problems including cerebral palsy (a lifelong problem with body movements) and/or mental
retardation.
It is not known why some pregnancies are affected by FMH and others are not. It is thought
that FMH may occur more frequently now than in the past, but no one knows why. If identified
early, FMH is readily treatable by blood transfusion of the baby before or after birth
and/or early delivery. Current laboratory testing for FMH is difficult and expensive. There
is great need identify high risk patients early in pregnancy in order to treat the condition
before the baby gets sick.
Approach: Five hundred women will be asked to participate in the study at the time they are
admitted to the Mount Sinai labor floor for delivery at term. After birth, newborns of study
mothers will be tested for anemia. Mothers of anemic babies will donate blood for
confirmation of FMH by established laboratory methods as well as for development of a new
laboratory screening protocol. All mothers will provide medical, social, environmental, and
full pregnancy history. Risk factors for FMH will be identified by statistical analysis of
this information.
Description:
1. Introduction:
The objective of this study is to identify candidate clinical predictors of
fetomaternal hemorrhage (FMH) and to devise a screening strategy to identify
pregnancies affected by the condition before the fetus is compromised by severe anemia.
Fetomaternal hemorrhage is a condition in which the placental barrier fails and the
fetus "bleeds" into the maternal circulation. When functioning correctly, the placenta
allows transfer of nutrients and waste between the mother and fetus while keeping the
cellular components of blood separate. It is not uncommon for small amounts of fetal
blood to reach the maternal circulation without adverse effect on the fetus. In fact,
mild FMH can be detected in up to 75% of normal pregnancies. In approximately 3 in 1000
pregnancies, however, the volume of fetal blood transferred to the mother causes
clinically-significant anemia in the fetus. Fetal anemia can cause significant
morbidity and mortality. In fetuses who survive severe anemia, life-long disability is
common.
Diagnosis of FMH is most commonly made after an adverse fetal or neonatal outcome has
occurred, indicating the need for testing. Early risk factors for FMH are unknown.
Clinical predictors of FMH have been suggested, but have not been born out in
retrospective study. Current standard of care testing for FMH, the Kleihauer-Betke (KB)
acid elution test, is labor-intensive and time-consuming, and therefore expensive.
Additionally, the KB test is observer-dependent and can be significantly affected by
variations in sample preparation. No screening protocol for FMH with automated
laboratory testing appropriate for use in the general pregnant population exists.
Specific Aim: To identify an appropriate laboratory protocol for diagnosis of
fetomaternal hemorrhage by prospective study and to pilot a study intended to determine
early clinical predictors of mild fetomaternal hemorrhage.
2. Background and Significance Fetomaternal hemorrhage occurs when the normal flow of
blood within the placenta is disrupted and the cellular components of fetal blood cross
into the maternal circulation. A healthy placenta permits transfer of dissolved
substances between the mother and fetus while keeping the cellular components of the
two circulations separate. It is common for the placental filter to "leak" during
normal pregnancies, resulting in transfer of small volumes of fetal whole blood into
the maternal bloodstream. Volumes of fetal blood under 1 mL can be detected in up to
75% of pregnancies. This volume of blood loss is thought to be clinically insignificant
to the fetus. Severe FMH with a large volume of blood transfer from the fetus to the
mother is less common. Although the exact volume of blood transfer needed to classify
FMH as "severe" is debated, it is clear that significant acute or chronic hemorrhage
leads to adverse perinatal outcome. Estimates of the incidence of severe FMH vary,
primarily because no comprehensive epidemiologic studies of the condition exist.
Estimates based on screening for Rh alloimmunization in Rho(d) negative women suggest
that severe FMH occurs in 1-3 of 1000 pregnancies, , but accounts for almost 14% of
otherwise unexplained fetal deaths. Large volume FMH can result in severe fetal anemia.
Although fetal anemia can manifest in utero with recognized hydrops fetalis or
intrauterine growth restriction, more commonly there are no outward signs of
FMH-related severe anemia before stillbirth, fetal distress at delivery, or neonatal
critical illness. The immediate sequelae of severe FMH for neonates that survive
delivery include devastating illnesses such as persistence of the fetal circulation,
hypovolemic shock, and hypoxic-ischemic encephalopathy. Ultimate detrimental outcomes
include high rates of mental retardation, cerebral palsy, neurologic devastation, and
death. Despite these consequences, and a suspected recent rise in incidence of severe
FMH, early predictors of severe FMH remain unknown and no cause of hemorrhage can be
identified in over 80% of cases. Maternal or pregnancy risk factors appropriate for
early screening or diagnosis of severe FMH in the general pregnant population have not
been elucidated. , If identified prior to delivery, fetal anemia from FMH can be
successfully managed by intrauterine fetal transfusion and delivery prior to the onset
of labor. , Outcomes following treatment of fetal anemia with fetal transfusion, even
repeated fetal transfusion, are encouraging. Thus, empirical predictors of FMH would
offer immediate promise for improving clinical outcomes. The social and financial cost
of both initial intensive care and long-term chronic care for children with cognitive
and motor disability resulting from FMH is substantial. There is considerable need for
early risk identification and a practical screening strategy to mitigate the human and
economic costs of severe FMH.
Validated clinical predictors of FMH include decreased fetal movement, sinusoidal fetal
heart rate tracing, and hydrops fetalis. , These markers are not useful for prevention of
adverse events stemming from FMH as each signifies established severe fetal anemia. Maternal
characteristics purported to increase the risk of FMH include hypertension, placental
abruption, substance abuse, and trauma. These were evaluated in retrospective study of
stillbirths in a 1999 study. None were significantly associated with FMH. Clinical
characteristics with suspected association with FMH were evaluated in a 2004 study of
Rho(d)-negative women. Only twin gestation was significantly associated with FMH. It is
unknown whether the incidence of FMH in multiple gestations is higher than those in
singleton pregnancies. No prospective or retrospective epidemiologic survey of FMH in the
general pregnant population has been published. No epidemiologic description over the
spectrum (mild, moderate, severe) of FMH exists. No series of placental pathology drawn from
cases of FMH has been completed.
Currently, even if at-risk pregnancies worthy of screening are identified prior to
devastating outcome, laboratory testing for FMH is not straightforward. The most commonly
used laboratory test to detect and quantify the volume of fetal blood in the maternal
circulation is the Kleihauer-Betke (KB) acid elution test. The reagents for the KB test are
relatively inexpensive, but the test is operator-dependent and labor-intensive making it an
unappealing candidate for use in expanded screening. The test is based on the fact that an
acid eluent will lyse fragile maternal erythrocytes while fetal hemoglobin stabilizes fetal
erythrocytes. Maternal blood is obtained and exposed to the acid eluent. Remaining intact
fetal cells are stained and counted. Estimates of maternal and fetal total blood volume are
then used to calculate the volume of FMH. The KB test is influenced by sample handling, is
technician-dependent, and can be inaccurate if patient blood volumes or blood counts are not
similar to standard estimates used. It is hypothesized that using maternal height and weight
to estimate maternal blood volume for each individual tested can improve the accuracy of KB
testing. This method is not widely practiced and has not been validated in the general
pregnant population. Such a "corrected KB" warrants further examination.
An additional test that has been explored to avoid some of the pitfalls of KB testing is
evaluation of maternal serum alpha-fetoprotein (AFP) levels. The concentration of
fetal-derived AFP in the maternal circulation can be used to estimate volume of FMH. As AFP
levels vary during and across pregnancies, however, AFP testing is most useful when multiple
levels obtained over the course of pregnancy are compared. The vast majority of pregnant
women in the United States undergo AFP testing in the second trimester as part of the "quad"
or "triple" screening tests for fetal aneuploidy and neural tube defects. Although
theoretically viable, no published reports document the use of this second-trimester AFP
baseline level for detecting FMH following additional AFP testing closer to term.
The most accurate measurement of FMH developed to date is achieved with flow cytometry (FC)
using antibodies specific to the hemoglobin of fetal cells. Cells must be made permeable
chemically to permit exposure of the FC antibody to intracellular hemoglobin prior to
analysis. Additionally, FC using monoclonal antibodies to fetal hemoglobin has only been
documented to reliably diagnose FMH >0.1% in the general clinical population. This is less
sensitive than the KB test when performed under standard clinical conditions. Finally, as
currently practiced, FC using antibodies to fetal hemoglobin is time-consuming and
expensive. This test is therefore not appropriate for large-scale screening at this time.
No series of placental pathology related to FMH has been published. As FMH is thought to be
a disease of placental failure, systematic review of placentas of pregnancies affected by
FMH could illuminate the pathogenesis of this type of placental abnormality.
Fetomaternal hemorrhage is an under-studied disease entity with significant associated
morbidity and mortality. This study aims to identify early clinical predictors of the
disease and to develop an automated screening strategy appropriate for wide-spread use.
Standard epidemiologic techniques will be used in combination with adaptation of existing
laboratory techniques to fully address this problem.
4. Methods: To address these Specific Aims, we propose (1) to test our automated laboratory
testing procedures for FMH and (2) to pilot a protocol using the novel automated laboratory
testing procedures to identify neonate-mother pairs affected by FMH for identification of
early clinical predictors of FMH. This is a pilot prospective nested case-control study.
In collaboration with the Red Cell Physiology laboratory at the New York Blood Center, we
have developed a prototype automated assay for FMH using flow cytometry facilities and
reagents available at the New York Blood Center.
A convenience sample of women admitted to the Mount Sinai Medical Center labor floor will be
selected for this pilot study. Eligible women will be those admitted for term delivery
(delivery between 37 0/7 and 41 6/7 weeks from the last menstrual period). Mothers carrying
fetuses with known fetal anomaly will be excluded. Women with pre-eclampsia or anomaly of
placental implantation (accreta, abruption) will be included, as these conditions may place
the pregnancy at risk for FMH but do not currently indicate pregnancy surveillance for fetal
anemia. Multiple gestations will be included. Women unable to complete the consent process
due to likely precipitous delivery, severe labor discomfort, or fetal distress requiring
immediate obstetric intervention will be excluded. Each woman will be enrolled in the study
prior to the required antepartum blood draw and/or IV placement, such that a study blood
specimen can be obtained with clinically indicated phlebotomy. Study specimens will be
banked for batch testing those from pregnancies of interest.
Not all women who donate blood as part of this study will have their blood tested for signs
of FMH. Although 5mL of blood will be drawn from eligible consenting women prior to
delivery, only specimens of those selected for our stratified sample population based on
neonatal hematocrit (HCT) will be tested. All maternal blood drawn will be fixed and/or
frozen immediately after phlebotomy for later batched analysis.
Eligible neonates of consenting mothers will have 0.5mL (approximately 3 drops) of blood
collected for determination of hematocrit at the time of the required blood draw for the New
York State Newborn Screening Program on day two of life. Eligibility will be determined by
the absence of a clinical history other than FMH to explain neonatal anemia. Clinical
indicators of anemia unrelated to FMH that will be treated as exclusion criteria include
hemolytic disease (due to known Rh, Kell, etc sensitization, or Coombs test positive), known
significant cord compression at the time of delivery (prolapsed cord, tight cord around the
neck or body), twin-to-twin transfusion syndrome recognized as anemia in the donor twin and
polycythemia in the recipient twin, and known neonatal blood loss (unclamped cord,
laceration). Neonates who have hematocrit measured within the first 48 hours of life for a
clinical indication will not have additional blood drawn for this study; results of
clinically-indicated samples will be used. All newborns of enrolled mothers will have HCT
tested. All mother-baby pairs with demonstrated neonatal anemia (HCT ≤ 50%) will be included
in the pilot cohort. In the case of multiple gestation, all siblings will be included for
analysis if any demonstrate anemia. Twenty-five percent of neonates with HCT over 50% will
also be included in the pilot cohort (every fourth study subject with HCT > 50%). This
sampling strategy will allow evaluation over the spectrum of newborn blood counts,
over-representing those with anemia.
Maternal blood of study cohort subjects undergo multiple tests for FMH including FC, AFP,
and KB. Flow cytometry will be performed at the New York Blood Center. Determination of AFP
level and KB testing will be performed by the Mount Sinai Hospital laboratory.
Maternal blood samples will be appropriately fixed at the time of collection, then frozen
and batch tested weekly. Any clinically-indicated testing for FMH will be performed
independently on blood specimens obtained post-partum, and will not be included in our
analysis. All available first trimester AFP testing results will be obtained for cohort
mothers. Anthropometric data will be gathered to correct testing results for estimated
maternal blood volume based on maternal height and weight. Accuracy of various testing
methods at predicting volume of fetal blood in the maternal circulation will be determined
by comparison with the KB specimens corrected for actual rather than estimated maternal
blood volume.
All placentas of study subjects will be fixed and held by the pathology laboratory for
standard one-week duration post-partum. Thirty pregnancies chosen by stratified sampling
based on severity of FMH will have the pregnancy placenta(s) subjected to a full pathologic
exam. Standard examination techniques including gross and microscopic exam will be used.
After study cohort mothers have been identified, we will collect medical, social,
environmental, and full pregnancy history via directed interview. Elements of particular
interest will include those hypothesized in the literature to be associated with FMH.
Factors suspected in the literature as being associated with FMH include maternal
hypertension during pregnancy, maternal pre-eclampsia, maternal trauma during pregnancy,
placental abruption, and multiple gestation. The survey tool will investigate these elements
of the maternal history as well as take a broad survey of the environmental and social
history in an attempt to identify unrecognized clinical predictors of FMH. Maternal subjects
will be interviewed during the post-partum stay in the hospital. Data will be used for
identification of early risk factors of FMH.
Risk factors will be identified through logistic regression modeling with a multilevel
outcome in which FMH is coded as absent (referent), mild, or severe, generating separate
odds ratios for mild and severe FMH. The alternate laboratory approaches to identification
of FMH will be compared by examining agreement using a kappa statistic, and quantifying
agreement across categories.
