Cytomegalovirus and Epstein–Barr virus may be associated with some cases of cerebral palsy

Objective: Intrauterine infection is a risk factor for cerebral palsy. Previous work reported a high frequency of viral DNA in newborn screening cards from cerebral palsy cases and controls possibly due to contamination. Methods: Retrospective case-control study using improved methodologies to minimize contamination during PCR-based detection of viral DNA sequences. Newborn screening cards of 339 Caucasian children with cerebral palsy and 594 controls were examined. Viruses tested were herpes simplex viruses 1 and 2 (HSV1 and 2), varicella zoster virus (VZV), Epstein–Barr virus (EBV), cytomegalovirus (CMV), human herpes viruses 6, 7 and 8 (HHV6, HHV7 and HHV8), and parvovirus B19. Genotyping was performed on DNA extracted from dried blood spots. Results: CMV and EBV were detected in 5 (1.5%) and 3 (0.9%) of 339 cases, respectively, but not in controls (p?=?0.047 and 0.006). Frequencies of detection of the other viruses examined were similar for cases and controls. DNA from at least one of the nine viruses tested was found in 4.4% of cases and 3.1% of controls [OR 1.4 95% CI (0.71–2.76)]. Conclusion: Evidence of congenital viral infection was uncommon in cases of cerebral palsy and controls. However, CMV and EBV were significantly associated with cerebral palsy.     

Detection of viral deoxyribonucleic acid in amniotic fluid: association with fetal malformation and pregnancy abnormalities

OBJECTIVE: To first test the hypothesis that the presence of viral nucleic acid in amniotic fluid (AF) is associated with an abnormal pregnancy outcome, and second, to determine if the overall rate of polymerase chain reaction (PCR) positivity and the distribution of virus types vary geographically. STUDY DESIGN: Cytomegalovirus (CMV), parvovirus B19, adenovirus, enterovirus, herpes simplex virus, Epstein-Barr virus, and respiratory syncytial virus nucleic acids were sought in 423 AF samples obtained for clinical indications: 284 from the East Coast (EC) and 139 from the Midwest (MW). RESULTS: Gestational age at sampling was 19.1 weeks for EC and 20.1 weeks for MW. 13.5% of karyotypically normal singleton pregnancies (57/423) had a positive AF PCR. 11% of AF PCR from the EC while 18% of AF PCR from the MW were positive (p = 0.06). The most commonly detected viruses were adenovirus (77%), enterovirus (12%), and CMV and parvovirus B19 (5% each). Twenty-four percent of sonographically abnormal pregnancies (33/136) had a positive AF PCR compared to only 8.4% (24/287) of normal pregnancies (p < 0.001). CONCLUSION: A positive AF PCR is associated with an increased rate of fetal structural malformations, intrauterine growth restriction, hydrops and other fetal abnormalities. There were no significant geographic differences in the incidence of AF viral PCR positivity.     

Molecular detection of HHV1-5, AAV and HPV in semen specimens and their impact on male fertility

Viral infections have been considered as possible destructive factors that influence male fertility. The aim of this study was to determine the prevalence of human herpes viruses 1-5 (HHV1-5), adeno associated virus (AAV) and human papilloma virus (HPV) in semen and whether these influence semen quality. DNA extraction was performed using phenol–chloroform protocol, then three different nested-PCRs were done to detect HHV1-5, AAV and HPV DNAs in the semen samples. Of 145 samples, 66 (45.5%) were positive at least for one of the viruses. The genome detection rate of HSV1/2, VZV, EBV, HCMV, AAV and HPV were zero, 2.8%, zero, 1.4%, 27.6% and 19.3%, respectively. Of 66 positive samples for these viruses, 6 (4.1% of all samples) were positive for two viruses simultaneously. Here no association was found between variations in semen parameters related to fertility and detection of VZV, HCMV, AAV and HPV DNA in semen samples. It should be noted that the prevalence of different viruses in semen, and their relevance to male infertility, differs significantly due to the genome extraction and amplification methods or due to a real variation between study populations and geographical regions.     

Detection of parvovirus B19, cytomegalovirus and enterovirus infections in cases of intrauterine fetal death

AIMS: Maternal infections with parvovirus B19, cytomegalovirus (CMV) and enterovirus have been associated with intrauterine fetal death (IUFD), but the incidence of these infections is not clear. This prospective study was conducted to estimate this incidence. METHODS: A prospective study of 38 months was conducted on cases of IUFD referred to Huddinge University Hospital, Stockholm, Sweden. Placental biopsies, fetal blood and amniotic fluid were collected from cases of IUFD (n=52). Placental biopsies from normal pregnancies at term (n=53) were used as controls. These tissues were examined for parvovirus B19 DNA, CMV DNA and enterovirus RNA using polymerase chain reaction (PCR). Maternal viral serology was measured in 46 cases and virus isolation for enterovirus in maternal stool samples was performed in 31 cases. RESULTS: Viral nucleic acid was recovered in at least one tissue sample from six cases of fetal death (parvovirus B19 in two cases, CMV in three and enterovirus in one), while all placental biopsies from controls were found negative. Serological signs of primary maternal infection were found in two of the cases, and virus isolation for enterovirus was negative in all samples examined. CONCLUSION: Parvovirus B19, CMV and enterovirus may be considered as etiologic agents in cases of fetal death. PCR on placental and/or fetal tissue improves diagnostic accuracy for these infections.     

应用套式聚合酶链反应技术检测孕妇与胎儿巨细胞病毒感染

目的评价套式聚合酶链反应技术（ＮＴ－ＰＣＲ）加限制酶分析在孕妇与胎儿人巨细胞病毒（ＨＣＭＶ）感染检测中的应用价值。方法应用ＮＴ－ＰＣＲ及限制酶分析、病毒分离、电镜观察和特异性抗体测定,对各孕期孕妇３６７例的外周血、部分孕妇脐血及死胎组织进行ＨＣＭＶ检测。结果孕早、中、晚期ＨＣＭＶ阳性检出率分别为８．６％、１．６％及７．０％,ＮＴ－ＰＣＲ检出率（４．９％）高于病毒分离（３．０％,Ｐ＜０．０２５）。６份ＨＣＭＶＤＮＡ阳性母血中,３份配对脐血ＨＣＭＶＤＮＡ也阳性。其中２对ＮＴ－ＰＣＲ、病毒分离及特异性ＩｇＭ、ＩｇＡ均阳性,１对ＮＴ－ＰＣＲ、病毒分离、特异性ＩｇＡ阳性,而ＩｇＭ阴性。２８例死胎组织中,发现１例死胎肺组织ＨＣＭＶＤＮＡ阳性。结论ＮＴ－ＰＣＲ能提高诊断ＨＣＭＶ感染的特异性与敏感性     

Viral infections in pregnancy

Viral infections are a common complication of pregnancy and in some cases, can have profound effects for the unborn fetus. The human herpesvirus family is composed of large, enveloped DNA viruses that have close structural similarity. The family includes the herpes simplex viruses types 1 and 2, varicella zoster virus, Epstein Barr virus, cytomegalovirus (CMV), and human herpes viruses types 6, 7 and 8. These viruses all share the ability to establish latency and reactivate at a later time. Structural fetal abnormalities can result from intrauterine infection and transmission of the infection during the pregnancy or at the time of delivery can result in important neonatal disease. Human parvovirus B19 is a DNA virus with strong tropism for erythroid precursors and infection during pregnancy can result in fetal hydrops and stillbirth. The causative agents of hepatitis are hepatotropic viruses termed hepatitis A, B, C, D (deltavirus) and E. All except hepatitis B virus are RNA viruses. Vertical transmission of maternal infection with hepatitis B and C can result in significant long term sequelae.     

Seroprevalence, incidence of prenatal infections and reliability of maternal history of varicella zoster virus, cytomegalovirus, herpes simplex virus and parvovirus B19 infection in South-Western Finland

Objective   To study seroprevalence and incidence and fetal transmission of varicella zoster virus (VZV), cytomegalovirus (CMV), herpes simplex virus (HSV) types 1 and 2 and parvovirus B19 infections during pregnancy and to evaluate the reliability of maternal past history of VZV, HSV and parvovirus infections.
Design   Prospective study of parturient women.
Setting   South-Western Finland.
Participants   Five hundred and fifty-eight parturient women.
Methods   IgG and IgM antibodies against VZV, CMV, HSV-1 and -2, and parvovirus B19 were measured from maternal serum in the first trimester and at delivery and from cord serum, mother's own information of her past infections was compared with her serological status.
Main outcome measures   Seroprevalence, seroconversions and fetal transmission of VZV, CMV, HSV and parvovirus B19, reliability of maternal history of VZV, HSV and parvovirus B19.
Results   Seroprevalences were 96.2% for VZV, 56.3% for CMV, 54.3% for HSV, 46.8% for HSV-1, 9.3% for HSV-2 and 58.6% for parvovirus B19. Parity was associated with CMV seropositivity, maternal age differed only between HSV-2 seropositive and seronegative women, while area of residence (urban or rural) had no effect. Six seroconversions were observed: two VZV, one CMV and three parvovirus infections. No cases of primary HSV infections occurred. Fetal transmission was observed in two cases of parvovirus infection. No infants with anti-CMV IgM antibodies were born to CMV immunised women. False positive history of chickenpox was given only by 1.5% of the women, history of herpes infections was less reliable, and history of parvovirus infection was unreliable.
Conclusions   Seroprevalence and the risk of viral infections during pregnancy cannot be extrapolated from one pregnant population to another.     

Maternal serum concentrations of the chemokine CXCL10/IP-10 are elevated in acute pyelonephritis during pregnancy

Objective. Acute pyelonephritis is one of the most frequent medical complications of pregnancy, as well as a common cause of antepartum hospitalization. Interferon (IFN)-γ inducible protein, CXCL10/IP-10, is a member of the CXC chemokine family with pro-inflammatory and anti-angiogenic properties. The purpose of this study was to determine whether maternal serum concentrations of CXCL10/IP-10 change in patients with acute pyelonephritis during pregnancy.

Study design. This cross-sectional study was conducted to determine the difference in maternal serum concentrations of CXCL10/IP-10 in pregnant women with acute pyelonephritis (N = 41) and normal pregnant women (N = 89). Pyelonephritis was defined in the presence of a positive urine culture, fever, and maternal clinical signs; blood cultures were performed in 36 cases. Maternal serum concentrations of CXCL10/IP-10 were measured by a sensitive immunoassay. Non-parametric statistics were used for analysis.

Results. (1) The median serum concentration of CXCL10/IP-10 in pregnant patients with pyelonephritis was significantly higher than in normal pregnant women (median 318.5 pg/mL, range 78.8–2459.2 vs. median 116.1 pg/mL, range 40.7–1314.3, respectively; p < 0.001); (2) maternal median serum concentrations of CXCL10/IP-10 did not differ significantly among patients with acute pyelonephritis with and without bacteremia (positive blood cultures: median 362.6 pg/mL, range 100.2–2459.2 vs. negative blood cultures: median 298.9 pg/mL, range 108.5–1148.7, respectively; p = 0.3).

Conclusions. Pyelonephritis in pregnant women is associated with an increased maternal serum concentration of the chemokine CXCL10/IP-10.     

Maternal serum concentrations of the chemokine CXCL10/IP-10 are elevated in acute pyelonephritis during pregnancy.

OBJECTIVE: Acute pyelonephritis is one of the most frequent medical complications of pregnancy, as well as a common cause of antepartum hospitalization. Interferon (IFN)-gamma inducible protein, CXCL10/IP-10, is a member of the CXC chemokine family with pro-inflammatory and anti-angiogenic properties. The purpose of this study was to determine whether maternal serum concentrations of CXCL10/IP-10 change in patients with acute pyelonephritis during pregnancy. STUDY DESIGN: This cross-sectional study was conducted to determine the difference in maternal serum concentrations of CXCL10/IP-10 in pregnant women with acute pyelonephritis (N = 41) and normal pregnant women (N = 89). Pyelonephritis was defined in the presence of a positive urine culture, fever, and maternal clinical signs; blood cultures were performed in 36 cases. Maternal serum concentrations of CXCL10/IP-10 were measured by a sensitive immunoassay. Non-parametric statistics were used for analysis. RESULTS: (1) The median serum concentration of CXCL10/IP-10 in pregnant patients with pyelonephritis was significantly higher than in normal pregnant women (median 318.5 pg/mL, range 78.8-2459.2 vs. median 116.1 pg/mL, range 40.7-1314.3, respectively; p < 0.001); (2) maternal median serum concentrations of CXCL10/IP-10 did not differ significantly among patients with acute pyelonephritis with and without bacteremia (positive blood cultures: median 362.6 pg/mL, range 100.2-2459.2 vs. negative blood cultures: median 298.9 pg/mL, range 108.5-1148.7, respectively; p = 0.3). CONCLUSIONS: Pyelonephritis in pregnant women is associated with an increased maternal serum concentration of the chemokine CXCL10/IP-10.     

HIV detection in amniotic fluid samples. Amniocentesis can be performed in HIV pregnant women?

OBJECTIVES: To assess amniotic fluid (AF) HIV contamination as a marker of intrauterine HIV infection and to evaluate amniocentesis as a risk factor for vertical HIV transmission when the mother was under antiretroviral treatment. STUDY DESIGN: Three hundred and sixty-six HIV pregnant women were included in the study. AF was obtained from three groups of patients: (a) genetic amniocentesis before 1997 (n=11); (b) amniocentesis a few days before the delivery day (n=18); and (c) AF collected on delivery (n=38). An univariate study was conducted to analyze amniocentesis as a risk factor of HIV transmission (groups a and b). Groups b and c were recruited after 1997; these patients were under combined antiretroviral treatment, they were studied to relate AF HIV contamination with fetal infection and maternal blood viral load at delivery (n=56). RESULTS: From 1984 to 1996, before antiretroviral therapy use in HIV pregnant women, transmission rate was 17%. In the group of patients who underwent genetic amniocentesis (group a) it was 30% (3/10) versus 16.2% (40/247) for patients without amniocentesis. Between 1997 and 2000 transmission rate was 3%. In group b it was 0% (0/18) when amniocentesis was done versus 3.7% (3/81) if no amniocentesis was done (no statistical differences). AF virus was undetectable in all samples (n=56) and no newborn infection was observed after the follow up. CONCLUSIONS: Amniotic fluid virus load was undetectable when maternal antiretroviral therapy was used, even if the virus was detectable in maternal blood samples. This finding raises the possibility to perform amniocentesis, when it is indicated, to provide the mother with an adequate antiretroviral treatment.     

A point of care test for the determination of amniotic fluid interleukin-6 and the chemokine CXCL-10/IP-10

Objective: Intra-amniotic inflammation is a mechanism of disease implicated in preterm labor, preterm prelabor rupture of membrane, cervical insufficiency, a short cervix, and idiopathic vaginal bleeding. Determination of interleukin (IL)-6 with immunoassays has been proven for more than two decades to be an excellent method for the detection of intra-amniotic inflammation. However, assessment of IL-6 for this indication has been based on immunoassays which are not clinically available, and this has been an obstacle for the implementation of this test in clinical practice. It is now possible to obtain results within 20?min with a point of care (POC) test which requires minimal laboratory support. This test is based on lateral flow-based immunoassay. The objective of this study was to compare amniotic fluid (AF) IL-6 and interferon-γ – inducible protein 10 (IP-10 or CXCL-10) concentrations determined using lateral flow-based immunoassay or POC test and standard enzyme-linked immunosorbent assay (ELISA) techniques.

Material and methods: AF samples were collected from patients with singleton gestations and symptoms of preterm labor (n?=?20). AF IL-6 and IP-10 concentrations were determined by lateral flow-based immunoassay and ELISA. Intra-amniotic inflammation was defined as AF IL-6?≥?2.6?ng/ml. AF IL-6 and IP-10 concentrations between two assays were compared.

Results: (1) Lateral flow-based immunoassay POC AF IL-6 and IP-10 test results were strongly correlated with concentrations of this cytokine/chemokine determined by ELISA (Spearman's ρ?=?0.92 and 0.83, respectively, both p?<?0.0001); (2) AF IL-6 concentrations determined by the lateral flow-based immunoassay test were, on average, 30% lower than those determined by ELISA, and the median difference was statistically significant (p?<?0.0001); and (3) in contrast, AF IP-10 concentrations determined by the lateral flow-based immunoassay test were, on average, only 7% lower than those determined by ELISA, and the median difference was not statistically significant (p?=?0.81).

Conclusion: AF IL-6 and IP-10 concentrations determined using a lateral flow-based immunoassay POC are strongly correlated with concentrations determined by conventional ELISA. This justifies further studies about the diagnostic indices and predictive values of this POC test.     

Can we use neopterin as marker of viral infections in pregnant women with symptoms of imminent spontaneous abortion?

Purpose.?To find out if determination of neopterin can be used for the detection of viral infections in pregnant women with symptoms of imminent spontaneous abortion.

Methods.?Eighty-eight pregnant women with symptoms of imminent spontaneous abortion (investigated group) and 88 healthy pregnant women were evaluated (control group). Neopterin level and IgM and IgG antibodies for eight viruses in the blood were determined.

Results.?Parvo B19 virus and elevated neopterin values were found in significantly higher number in investigated group than in control. There was no correlation between women with acute ParvoB19 infection and elevated neopterin level.

Conclusions.?The determination of neopterin in the sera cannot be used for screening of viral infections in pregnancy.     

Prenatal diagnosis of human cytomegalovirus by culture and polymerase chain reaction: 98 pregnancies leading to congenital infection.

Human cytomegalovirus (HCMV) is the most common cause of viral intra-uterine infection. The experience with prenatal diagnosis remains limited and is based on few reports of small numbers of cases. It is thus difficult to compare the accuracy of the different tests because the groups studied were small and heterogeneous. We describe here our experience on a series of 98 pregnancies leading to HCMV congenital infection, among which 71 have been tested by amniotic fluid (AF) sampling followed by culture and/or polymerase chain reaction (PCR). Independently of the delay between AF sampling and the first HCMV IgM positive result, the mean sensitivity of both culture and PCR was around 70 per cent. The best sensitivity (95.5 per cent) was obtained after a delay > or = 6 weeks in late pregnancy (> or = 23 weeks). The present study demonstrated clearly that the delay between AF puncture and the presumed date of seroconversion is more important for sensitivity than the technique used for the diagnosis (PCR or culture). However, even in the best diagnostic conditions, negative results of HCMV culture or PCR in AF cannot formally exclude intra-uterine infection.     

Fetale Virusinfektionen

Infections during pregnancy may adversely affect pregnancy outcome and child health. They may be associated with fetal death, preterm delivery, congenital defects, and an increased risk of perinatal morbidity and mortality. The risk of intrauterine transmission and fetal disease depends on the nature of the pathogen, type of maternal infection (primary, recurrent, or chronic infection), and gestational age at time of fetal infection. The most important viruses that may cause symptomatic fetal infections are human cytomegalovirus (CMV), human parvovirus B19 (B19V), varicella-zoster virus (VZV), and rubella virus (RV). Available preventive measures (e.g., active or passive immunization, exposure or postexposure prophylaxis) and treatment options as well as modern serological and virological diagnostics should be thoroughly used to minimize the risk of sequelae associated with prenatal viral infections..     

Amino acid concentrations in maternal plasma and amniotic fluid in relation to fetal insulin secretion during the last trimester of pregnancy in gestational and type I diabetic women and women with small-for-gestational-age infants

Free amino acid concentrations were determined in maternal plasma and amniotic fluid (AF) under standardized and unstressed conditions in four groups of women comprising 6 gestational and 13 type I diabetics, 10 women with small-for-gestational-age (SGA) infants, and 18 healthy control women between 36 and 39 weeks of gestation. Plasma values for branched chain amino acids (the sum of leucine, isoleucine and valine) did not differ significantly between the four groups. The corresponding values in AF were significantly higher (P less than 0.05) in the type I diabetic group and significantly lower (P less than 0.05) in the gestational diabetic group as compared to the control group. The mean AF C-peptide concentration was elevated but not significantly so in gestational (0.69 nmol/l) or type I diabetic (0.54 nmol/l) pregnancies and significantly lower (P less than 0.05) in women with SGA infants (0.28 nmol/l) as compared to the control group (0.38 nmol/l). There was a significant correlation between C-peptide in AF and branched chain amino acids in maternal plasma (r = 0.63; P less than 0.05) as well as to maternal blood glucose (r = 0.79; P less than 0.01) in the type I diabetic group, which merely suggests a greater beta cell reactivity to insulin secretagogues in offspring of diabetic mothers. The correlation between AF C-peptide and branched chain amino acids in maternal plasma was significantly inverse in women with SGA infants (r = -0.75; P less than 0.05). Both individual, branched chain, or total amino acid concentration in AF were unrelated to AF C-peptide.     

应用套式聚合酶链反应技术诊断人巨细胞病毒宫内感染

用套式聚合酶链反应（ＲＴ－ＰＣＲ）技术，对妊娠早、中期各３０例及妊娠晚期３１例孕妇进行母血、脐血及胎盘人巨细胞病毒（ＨＣＭＶ）ＤＮＡ测定。结果：早期妊娠妇女中，２０例ＨＣＭＶ阳性，其中８例发生宫内传播（绒毛组织中ＨＣＭＶ－ＤＮＡ阳性）；妊娠中期妇女中，２１例ＨＣＭＶ阳性，其中７例发生宫内传播（胎盘、脐血ＨＣＭＶ－ＤＮＡ阳性）；妊娠晚期妇女中，２３例ＨＣＭＶ阳性，其中１８例发生宫内传播。提示：套式Ｐ     

Viral hepatitis during pregnancy

Viral hepatitis is one of the most common liver diseases appearing during pregnancy. Prevention against hepatotropic viruses is restricted due to lack of vaccines being effective in induction of efficient immunization in the majority of these microorganisms. In general, there is no possibility of active immunization against hepatotropic viruses except type A and B viral hepatitis. An issue of viral hepatitis in pregnancy as an aspect of potential risk factor connected with infection of pregnant women and a fetus has been described in this paper. Furthermore, the most important topics in the field of the epidemiology, prophylaxis and possible treatment options of viral hepatitis A, B, C, D, E and G have been discussed. The newest reports of pregnant women lamivudine therapy as a preventive treatment against vertical transmission during delivery have been reviewed. Rarly diagnosed viral hepatitis caused by herpes simplex virus, cytomegalovirus, Epstein-Barr virus and adenoviruses have been characterized as well.     

Maternal cytokines and disease severity influence pregnancy outcomes in women with rheumatoid arthritis

Objectives: To assess the influence of maternal cytokine levels, disease activity and severity on preterm delivery, small for gestational age (SGA) and cesarean delivery in pregnant women with rheumatoid arthritis (RA).

Methods: A prospective study in 47 pregnant women with RA and 22 healthy pregnant controls. The main outcome measures were birth weight in relation to maternal serum levels of interleukin-6 (IL-6), interleukin-10 (IL-10), and RA activity and severity at three different time points: preconception and during the first and third trimesters.

Results: During the third trimester, IL-10 was detectable in 23.4% of patients with RA, IL-6 in 76.6%. Mean birth weight born to mothers with RA was higher when IL-10 level was high compared with low (p?=?0.001), and lower when IL-6 was high compared with low (p?=?0.035). Also increase in disease activity score-28 (in 60.1%, p?=?0.001), Health Assessment Questionnaire–Disability Index (in 87.5%, p?=?0.013), and pain score (56.9?±?11.4, p?=?0.003) associated with increased risk of SGA. High patient’s global scale was associated with unfavorable pregnancy outcome (preterm, SGA, and cesarean).

Conclusion: High maternal IL-10 levels are associated with higher birth weight and high IL-6 levels are associated with lower birth weight (SGA). Among women with RA, disease activity and severity are predictive of unfavorable pregnancy outcomes suggesting that better disease management early in the pregnancy could improve pregnancy outcomes.