Parvovirus B19 infection in pregnancy: Quantitative viral DNA analysis using a kinetic fluorescence detection system (TaqMan PCR) (original) (raw)
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Journal of clinical microbiology, 1996
Human parvovirus B19 infection in pregnancy represents a potential hazard to the fetus since fetal loss or fetal hydrops can occur. The risk of fetal loss due to transplacental B19 transmission has been evaluated in several studies using different diagnostic methods on maternal and fetal specimens. We analyzed the diagnostic value of virological and serological techniques on maternal serum, fetal cord blood, and amniotic fluid specimens obtained at the time of clinical diagnosis of fetal hydrops in 18 cases of B19 fetal hydrops. B19 DNA was detected by nested PCR, dot blot hybridization, and in situ hybridization assay. Anti-B19 immunoglobulin M and G antibodies were detected by immunoassays using recombinant B19 antigens. Our data suggest that for maternal sera, virological and serological methods have a complementary role in diagnosis, while for fetal specimens the in situ detection of B19 DNA in fetal cord blood is the most sensitive diagnostic system.
Prenatal diagnosis of human parvovirus B19 in nonimmune hydrops fetalis by polymerase chain reaction
International Journal of Gynecology & Obstetrics, 1993
OBJECTIVE: Nonimmune hydrops fetalis is a potentially lethal condition reflecting the clinical manifestation of several pathologic processes. Recently maternal infection by human parvovirus 819 has been reported to result in nonimmune fetal hydrops. We sought to develop a rapid and sensitive test to detect the presence of this agent in utero. STUDY DESIGN: Using a cloned isolate of the virus, we developed an assay based on enzymatic amplification of a segment of the human parvovirus 819 genome that allows direct detection of this agent in samples of fetal blood and amniotic fluid. RESULTS: The method detected as few as 100,000 genome equivalences and was specific for the viral genome alone. We used this assay to evaluate nine fetuses initially seen with nonimmune hydrops. Three cases were found to be positive for the human parvovirus 819 genome. CONCLUSION: The method is powerful in that it is rapid, sensitive, and simple. This assay may have general applicability in evaluation of nonimmune hydrops and in documentation of the natural history of fetal human parvovirus infections . (AMJ OaSTET GVNECOL 1992;167:461-6.)
Pathologie-biologie, 2009
The aim of the present study was to determine whether there is an association between Parvovirus B19 infection and hydrops fetalis setting in fetus and neonate. Twenty-nine samples were analyzed by three methods. Each sample was histologically examined for viral nuclear inclusions in fetal organs and placenta, then immunohistochemical study using Parvovirus B19 antibody that recognized the VP2 protein of the Parvovirus B19 capsid was done in tissue embedded in paraffin (lungs, liver, thymus, kidneys, heart and placenta). Nested-PCR analysis was done after DNA extraction from paraffin blocks and using specific primers of the Parvovirus B19 VP1 gene. Apparent causes of hydrops were eliminated such as metabolic diseases, cardiac failure or malformation. The standard histological study objects viral inclusion in one case (lung tissue). However, the immunohistochemical study was negative in all cases. Nested-PCR demonstrates the presence of the viral DNA in five cases. Our study demonstr...
Journal of Clinical Pathology, 1989
Attempts were made to detect human parvovirus B 19-DNA by in situ hybridisation and the polymerase chain reaction in placental and fetal tissues from a case of intrauterine fetal death. In the in situ hybridisation experiments radioactive and non-radioactive (labelled with 2-acetylaminofluorene, AAF) DNA probes were used. B19-DNA was detectable in paraffin wax embedded fetal tissue from the liver, heart, lung, brain and thymus. The resolution with the AAF-labelled probes was higher than with the radiolabelled DNA. Parvovirus B 19 DNA sequences were also detected in these tissues by an enzymatic in vitro amplification technique-the polymerase chain reaction. Amplification of a B 19-DNA sequence before detection increases the rapidity and sensitivity of detection.
Detection of Parvovirus B19 in Bad Obstetric History by Using Real Time PCR
Background Human parvovirus B19 (B19V) is a small single-stranded DNA virus. Infection during pregnancy can cause a variety of signs of fetal damage. The risk of adverse fetal outcome is increased if maternal infection occurs during the first two trimesters of pregnancy but may also happen during the third trimester.
Improved Diagnosis of Gestational Parvovirus B19 Infection at the Time of Nonimmune Fetal Hydrops
The Journal of Infectious Diseases, 2008
Background. In the diagnosis of parvovirus B19 infection, the detection of virus-specific IgG in the absence of virus-specific IgM is considered to indicate past immunity. Methods. We determined the diagnostic value of a high-quality B19 IgM EIA, compared with that of a VP1 IgG avidity EIA, a VP2 IgG epitope-type specificity (ETS) EIA, and real-time polymerase chain reaction (PCR) in the diagnosis of maternal B19 infection during nonimmune fetal hydrops. Results. Serum samples from 101 pregnant women with confirmed B19-induced fetal hydrops were collected at the time of invasive prenatal diagnosis. The samples were investigated for B19 IgM, VP1 IgG avidity, and VP2 IgG ETS. With the B19 IgM EIA, 78 women (77.2 %) showed positive results, 15 (14.9%) showed negative results, and 8 (7.9 %) showed equivocal results. According to the combined B19 IgG avidity and IgG ETS EIA results, only 5 (5%) of 101 women were classified as having past immunity. Available serum samples (n ϭ 24) that had nondiagnostic results in the antibody assays were further investigated by PCR. All were B19 DNA positive (mean load, 2.5 ϫ 10 4 genome equivalents/mL; range, 2.5 ϫ 10 3-7.8 ϫ 10 6). Conclusions. At the time of B19-induced hydrops, detection of B19 DNA in maternal blood had the best diagnostic sensitivity for identifying maternal B19 infection. However, given the long persistence of B19 DNAemia, supplementary measurement of VP1 IgG avidity and VP2 IgG ETS improves the precision of diagnosis and management of pregnant women affected by the B19 virus.
Intrauterine Parvovirus B19 infections may cause fetal anemia, non-immune hydrops fetalis or abortion. This study focuses on the pathogenic role of Parvovirus B19 in non-immune hydrops fetalis at Hacettepe University, a major reference hospital in Turkey. Twenty-two cases of non-immune hydrops fetalis were retrospectively selected out of a total of 431 hydrops fetalis specimens from the Department of Pathology archieves. Paraffine embedded tissue sections from placental and liver tissues from each case were evaluated by histopathology, immunohistochemistry, nested PCR and commercial quantitative Real-time PCR. Viral DNA was detected in placental tissues by Real-time PCR in 2 cases (2/22, 9.1%) where histopathology also revealed changes suggestive of Parvovirus B19 infection. No significant histopathologic changes were observed for the remaining sections. Nested PCR that targets the VP1 region of the viral genome and immunohistochemistry for viral capsid antigens were negative for all cases. As a result, Parvovirus B19 is identified as the etiologic agent for the development of non-immune hydrops fetalis for 9.1% of the cases in Hacettepe University, Turkey. Real-time PCR is observed to be an effective diagnostic tool for nucleic acid detection from paraffine embedded tissues. Part of this study was presented as a poster at XIIIth International Congress of Virology, San Francisco, USA (Abstract V-572).
The new microbiologica, 2016
To define diagnostic and prognostic markers of parvovirus B19 (B19V) fetal infection, two groups were investigated: (1) pregnant women with specific symptoms or contacts with symptomatic households (n=37); (2) mothers with pathological ultrasound findings and the relevant fetus at the time of prenatal diagnosis (n=16). In the first group, diagnosis of B19V infection was achieved using IgM detection in 29/37 (78.3%) of patients, while B19V DNA was detected in 36/37 (97.3%) of infected women. In the second group, intrauterine infection was investigated by amniocentesis (n=5), cordocentesis (n=3) or both (n=5). Median B19V DNA load in amniotic fluid was 8.2x107 copies/ml and in fetal blood was 2x109 copies/ml. Maternal blood was positive for B19V DNA (median 3.8x104 copies/ml) in 14/16 (87.5%) women examined. At time of fetal US investigation, all mothers were B19V IgG positive and B19V IgM were detected in 10/16 (62.5%), while fetal B19V IgG and IgM were detected in 1/8 (12.5%) and 5/...
Journal of Clinical Virology, 2006
Background: Over 95% of fetal complications (fetal hydrops and death) occur within 12 weeks following acute parvovirus B19 (B19) infection in pregnancy. Therefore, weekly fetal ultrasound monitoring is generally recommended for this time period. However, in the majority of women, typical symptoms of acute infection (rash or arthropathy) are absent, and during epidemics, B19 infection may be diagnosed incidentally by antibody screening of women at risk. Objective: To assess the diagnostic value of currently available molecular and serological methods for reliable diagnosis of primary B19 infection in pregnancy. Study design: Large panels of well-characterized acute-phase or convalescent sera were used to investigate the ability of a VP2 IgM EIA, a Light-Cycler-based B19-DNA PCR, a VP1-IgG avidity EIA and two VP2-IgG epitope-type specificity [ETS] EIAs to pinpoint the time of primary B19 infection in pregnancy. Results: The duration of low-level IgM positivity varied greatly (range 4-26 weeks). Samples collected within the first 2 weeks of infection showed high-level viremia (mean 1.75 × 10 8 geq/ml). During follow-up, low-level DNAemia (mean 9.7 × 10 4 geq/ml) persisted for at least 18 weeks in 91% (20/22) of patients. Considering the first 12 weeks after onset of disease the window of greatest risk for fetal complications, the "acute" phase was extended to cover this full period. In this case, performing the avidity and ETS-EIA sequentially, the positive predictive value was 100% in patients showing concordant avidity and ETS-EIA results. Conclusions: In the presence of low IgM titres and/or low-level DNAemia the use of supplementary serological assays such as VP1-IgG avidity EIA and VP2-ETS-EIA is advisable for restriction or avoidance of unnecessary fetal ultrasound examinations or invasive diagnostics; and in general for strengthening the reliability of B19 serodiagnosis of pregnant women.
Detection of Intrauterine Viral Infection Using the Polymerase Chain Reaction
Molecular Genetics and Metabolism, 1998
pressed cell-mediated immunity (1). Therefore, the Intrauterine viral infection commonly presents as fetus is at risk to become infected by transplacental nonimmune hydrops fetalis or intrauterine growth transmission during maternal viremia. Depending restriction. Cytomegalovirus (CMV) and parvovirus on the gestational age of the fetus, the involved viare commonly recognized causes of fetal infection rus, and the viral load, the spectrum of fetal involveusing serology and cultures. We used the polymerment may range from asymptomatic to severe, causase chain reaction (PCR) to evaluate the frequency of fetal viral infection and the associated clinical ing either fetal or neonatal death or long-term course and outcome. Specimens (amniotic fluid, fesequelae in survivors. Rubella, cytomegalovirus tal blood, pleural fluid, tissue) from 303 abnormal (CMV), herpes simplex virus (HSV), and the varipregnancies at risk for viral infection and 154 concella-zoster virus (VZV) are well-known teratogens trols were analyzed using primers for CMV, herpes (2-7). Congenital malformations after infection with simplex virus, parvovirus B19, adenovirus, enterocoxsackie virus and echo virus (cardiac, urological, virus, Epstein-Barr virus, and respiratory syncytial and gastrointestinal malformations), mumps (endovirus. Viral genome was detected in 144/371 samples cardial fibroelastosis), and influenza (central ner-(39%) or 124/303 patients (41%), with adenovirus (n vous system and heart defects) are also documented Å 74 patients; 24%), CMV (n Å 30 patients; 10%), and (8). Ultrasound findings during the second and third enterovirus (n Å 22 patients; 7%) most common. Only 4/154 (2.6%), unaffected control patients' sam-trimester of pregnancy consistent with intrauterine ples were PCR positive. We conclude that diagnosis growth restriction (IUGR), nonimmune hydrops, isoof fetal viral infection by PCR is common in abnorlated ascites, microcephaly, hydrocephaly, intramal pregnancies. Adenovirus and enterovirus may cranial, or intrahepatic calcification raise the suspicause fetal infection that have been previously uncion of a fetal viral infection affecting multiple orrecognized. ᭧ 1998 Academic Press gans (9).