Congenital HCMV infection: a collaborative and comparative study of virus detection in amniotic fluid by culture and by PCR (original) (raw)

Detection of HCMV DNA in placenta, amniotic fluid and fetuses of seropositive women by nested PCR

European Journal of Pediatrics, 2007

Human cytomegalovirus (HCMV) is the most common viral cause of intrauterine infection throughout the world. Its distribution patterns in different clinical samples are poorly understood. This study was performed to determine the frequency of CMV DNA positivity in maternal/fetus sera, placentas and amniotic fluid, together with maternal/fetus serology. Clinical specimens were obtained from 92 pregnant women who delivered by cesarean section. 98% of women and their neonates were HCMV IgG positive and 5.4% of these mothers were IgM positive, while no IgM was detected in neonates of IgM positive mothers. Among the IgG positive mothers, IgM was detected in 3.3% of their fetuses. 5.4% and 3.3% of maternal and fetal sera were HCMV DNA positive, respectively. The three neonates who were positive for HCMV DNA in sera were also positive for HCMV IgM and the PCR of their amnions was positive (p<0.0001). 9.8% of placenta samples and 4.3% of amniotic fluid specimens were positive for HCMV DNA while among these placenta samples, two amnions were PCR positive (p=0.046). Our results showed that there is not always a correlation between placenta and amnion infections. This may be due to reactivation of HCMV leading to placenta infection, as all affected placentas do not pass infection to fetuses and amniotic fluids. Detection of HCMV DNA in amnion and fetus plasma and the existence of fetus IgM against HCMV can also occur without clinical symptoms.

Polymerase chain reaction for prenatal diagnosis of congenital human cytomegalovirus infection

Journal of Medical Virology, 1995

The reliability of the polymerase chain reaction (PCR) for prenatal diagnosis of human cytomegalovirus (HCMV) infection was determined by retrospective testing of 35 amniotic fluids identified previously as positive or negative for HCMV by virus isolation. Amniocentesis was performed in 26 pregnant women with primary HCMV infection at 14-36 weeks gestation, 3-21 weeks after maternal infection. Blood samples were obtained from 20 fetuses for IgM determination and/or virus isolation. Amniotic fluid culture led to antenatal diagnosis of HCMV in 9 of the 13 infected fetuses (sensitivity 69.2%) with one case diagnosed at a second sampling. PCR was able to detect one additional infected fetus (10/13, sensitivity 76.9%). Nested PCR did not increase sensitivity of prenatal diagnosis. Three cases were not diagnosed by all the techniques employed. The specificity of virus isolation from and DNA detection by PCR in amniotic fluid was 100%. The negative predictive value for virus isolation from amniotic fluid was 76.5% and for DNA detection by PCR 81.2%, whereas the positive predictive value was 100% for both techniques. The results showed that neither approach can detect all cases of congenital HCMV infection prenatally, and that the time interval between maternal infection and sampling seems to be a major factor affecting the reliability of prenatal diagnosis.

The prevalence of congenital and perinatal cytomegalovirus infections among newborns of seropositive mothers

The Journal of Infection in Developing Countries, 2012

Introduction: This study aimed to determine the prevalence of congenital and perinatal human cytomegalovirus (HCMV) infections among newborns in two major neonatal intensive care units (NICU) in Bahrain. Methodology: One hundred newborns comprised of 84 preterm and 16 term babies admitted to the NICUs were enrolled in the study. During the first six weeks of life, urine and saliva was obtained from the babies weekly and serial breast milk samples were obtained from the mothers. Maternal serum HCMV IgG was measured. Virus isolation and detection was done by shell vial culture and nested PCR. Results: Maternal HCMV IgG-seropositivity was 100%. Eight HCMV infections were detected comprising of three congenital and five perinatal infections. Congenital HCMV infection was found in preterm (2/84; 1.9%) and term (1/16; 6.3%) babies. HCMV DNA was detected in breast milk samples obtained during the first 10 days postpartum from all mothers whose babies had congenital HCMV. Fortynine women provided breast milk samples between four and six weeks post-partum and HCMV DNA was detected in the breast milk of 11 women. Five (45.5%) of these eleven were mothers of babies with perinatal HCMV infection. There was no significant difference in the detection of HCMV using shell vial culture versus nested PCR method. Conclusion: The findings indicate occurrence of congenital and perinatal HCMV transmission in this setting of high maternal seropositivity. The use of shell vial culture and PCR amplification for HCMV screening in the NICU for rapid detection of infection during the early postnatal period is recommended.

Results of Routine Antenatal Screening for Cytomegalovirus at a Tertiary Center

Medeniyet Medical Journal

was reported to be between 0.3% and 2.4% 3. During pregnancy, whereas transplantal transmission of the virus spread was reported to be between 24% and 75% with the first infection of pregnant women, it was reported to be between 1% and 2.2% with non-primary infections 4. Additionally, while the rate of maternalfetal transmission is low in the first trimester, the rate of transmission to the fetus increases with advancing ABSTRACT Objective: Cytomegalovirus (CMV) is the most common viral infection. In this study, we discussed the results of pregnant women who underwent antenatal CMV screening in a tertiary center and the value of CMV antenatal screening. Methods: For this retrospective study, the data of pregnant patients with antenatal CMV screening test results between 2019 and 2022 were obtained from hospital records. CMV immunoglobulin M (IgM), CMV IgG, anti-IgG avidity test results, amniocentesis, CMV polymerase chain reaction (PCR), and the outcome of the babies were recorded. Results: A total of 31,912 CMV IgM and 26,969 CMV IgG tests were performed. CMV IgG seropositivity was observed in 78.99% of pregnant women, and 0.09% of the pregnant women were confirmed to have a positive CMV IgM test result. Pregnant women with positive IgM accompanying low avidity were referred to perinatology clinics for detailed ultrasonography and amniocentesis. Only 3 of the 44 pregnant women who underwent amniocentesis were confirmed to have positive CMV PCR testing. Conclusions: CMV screening should be preserved for pregnant women with ultrasonographic findings at high risk of congenital CMV infection.

Accuracy of amniotic fluid testing before 21 weeks' gestation in prenatal diagnosis of congenital cytomegalovirus infection

Prenatal Diagnosis, 1994

Cytomegalovirus (CMV) is the most common cause of intrauterine infection. Recent publications show amniocentesis to have an 81-100 per cent sensitivity in antenatal diagnosis after 21 weeks' gestation. Testing before 21 weeks' gestation is less well documented. We performed 36 amniocenteses between 14 and 20 weeks' gestation. The sensitivity was 45 per cent and the specificity 100 per cent. Implications and possible causes of this low sensitivity are discussed. KEY woms-Cytomegalovi, amniocentesis, prenatal diagnosis, fetus.

Prenatal diagnosis of congenital human cytomegalovirus infection

Prenatal Diagnosis, 1994

Fifteen fetuses at risk of congenital human cytomegalovirus (HCMV) infection underwent prenatal diagnosis at 16-30 weeks' gestation by a combination of amniocentesis and fetal blood sampling. HCMV was isolated from the amniotic fluid in six patients, but HCMV-specific IgM was detected in only three of them. Two of the nine neonates, who were delivered following a negative prenatal diagnosis, had congenital HCMV infection diagnosed by virus isolation in the urine. The interval from infection to prenatal testing was 3 and 4 weeks in the two false-negative cases and 2 7 weeks in the true-positive cases. Although timely testing for HCMV infection allows the option of termination of pregnancy, it may be flawed by false-negative results.

Real-Time PCR Quantification of Human Cytomegalovirus DNA in Amniotic Fluid Samples from Mothers with Primary Infection

Journal of Clinical Microbiology, 2002

A real-time PCR assay was developed to quantify human cytomegalovirus (HCMV) DNA in amniotic fluid (AF) samples collected from 30 pregnant women with primary HCMV infection as detected either from HCMV-immunoglobulin G (IgG) seroconversion or by the presence of HCMV-specific IgG and IgM associated with a low IgG avidity. Clinical information available for each case included ultrasonographic examination and fetal or newborn outcome. HCMV infection of fetuses or newborns was confirmed for the 30 studied cases. AF samples were subdivided into three groups. In group A (n ‫؍‬ 13), fetuses presented major ultrasound abnormalities, and pregnancy was terminated. In group B (n ‫؍‬ 13), fetuses had normal ultrasound findings, the pregnancy went to term, and the newborns were asymptomatic at birth. In group C (n ‫؍‬ 4), fetuses had no or minor ultrasonographic signs, and pregnancy was terminated. The HCMV DNA load values in AF samples were significantly higher in group A (median, 2.8 ؋ 10 5 genome equivalents [GE]/ml) than in group B (median, 8 ؋ 10 3 GE/ml) (P ‫؍‬ 0.014). Our findings suggest that HCMV load level in AF samples correlates with fetal clinical outcome but might also be dependent on other factors, such as the gestational age at the time of AF sampling and the time elapsed since maternal infection.

Clinical Predictive Value of Real-Time PCR Quantification of Human Cytomegalovirus DNA in Amniotic Fluid Samples

Journal of Clinical Microbiology, 2009

The aim of this study was to evaluate the diagnostic reliability and prognostic significance of the quantification of cytomegalovirus (CMV) DNA in amniotic fluid (AF). We retrospectively reviewed the results for 282 amniotic fluid samples that had been tested for CMV by a quantitative real-time PCR. We observed three cases in which no CMV genomes were detected in the AF but in which the children were nevertheless congenitally infected. Hence, we conclude that a negative result by PCR for CMV in AF cannot rule out the possibility of congenital infection. No false-positive PCR results were observed. A correlation between the CMV viral load in AF and the fetal and neonatal outcomes could not be demonstrated in our study. Instead, a correlation was found between the CMV viral load and the gestational age at the time of amniocentesis.

Comparison of conventional, immunological and molecular techniques for the diagnosis of symptomatic congenital human cytomegalovirus infection in neonates and infants

Indian Journal of Medical Microbiology, 2015

in the general population is between 50-70%, [3,4] the corresponding fi gure for the developing nations ranging from 70-100%. In India, this fi gure approaches 98-100%. [5] The rates of congenital infection in developed countries are about 0.6-0.7% of live births, whereas in the developing world, higher rates between 1-5% have been observed. [1,3] Congenital HCMV infection is asymptomatic in more than 90% of infected infants, whereas the remaining 10% show manifestations including microcephaly, small for gestational age (SGA), neonatal hepatitis, hepatomegaly, splenomegaly, chorioretinitis, cataract, thrombocytopenia, etc, Some of these symptomatic infants also develop sensorineural hearing loss (SNHL) in the ensuing few years. [2] A fraction of infants who were asymptomatic at birth also go on to develop complications like SNHL and neurodevelopmental delays in the next few years of their lives. In fact, it is now established that congenital HCMV infection is the leading cause of nonsyndromic SNHL in the developed world. [2] The diagnosis of congenital HCMV infection has traditionally been based on demonstration of the virus in urine by isolation in cell culture. Owing to the slow turnaround time for cell culture and its low sensitivity, demonstration of HCMV DNA in urine by PCR has gradually replaced virus isolation. [2] Demonstration of IgM antibodies to HCMV (anti-HCMV IgM) is a common diagnostic test in neonates or infants, which suggests current infection and supports the diagnosis. Other tests, utilising