Parvovirus B19 infection in five European countries: seroepidemiology, force of infection and maternal risk of infection.

We conducted a seroprevalence survey in Belgium, Finland, England & Wales, Italy and Poland on 13 449 serum samples broadly representative in terms of geography and age. Samples were tested for the presence of immunoglobulin G antibody using an enzyme immunoassay. The age-specific risk of infection was estimated using parametric and non-parametric statistical modelling. The age-specific risk in all five countries was highest in children aged 7-9 years and lower in adults. The average proportion of women of child-bearing age susceptible to parvovirus B19 infection and the risk of a pregnant women acquiring B19 infection during pregnancy was estimated to be 26% and 0.61% in Belgium, 38% and 0.69% in England & Wales, 43.5% and 1.24% in Finland, 39.9% and 0.92% in Italy and 36.8% and 1.58% in Poland, respectively. Our study indicates substantial epidemiological differences in Europe regarding parvovirus B19 infection.

Type specific seroprevalence of HSV-1 and HSV-2 in four geographical regions of Poland.

OBJECTIVE: To examine the type specific seroprevalence of herpes simplex virus (HSV) types 1 and 2 infections, stratified by age and gender, and associated risk factors for HSV-2 seropositivity in Poland. METHODS: 2257 serum samples of individuals from 15-65 years were randomly selected from serum banks in four different geographical regions of Poland, including the Zachodnio-pomorskie, Warminsko-mazurskie, Lubelskie, and Mazowieckie districts. Type specific serum antibodies to HSV-1 and HSV-2 were detected using HerpeSelect IgG ELISA tests. RESULTS: Overall prevalences of type specific HSV-1 and HSV-2 serum antibodies were 90.4% and 9.3%, respectively. Age standardised HSV-2 seroprevalence was higher in women (9.7%) than men (8.8%) (p = 0.06), and increased notably with age from 4% in 15-24 year olds to 12% in those aged 50-65 years. HSV-1 seroprevalence was consistently higher than HSV-2 seroprevalence in each specific age group, ranging from 74.5% in 15-24 year olds to 98.8% in 50-65 year olds. HSV-2 seroprevalence varied significantly by geographical region, with the highest prevalence in the Zachodnio-pomorskie district (12%). Significant multivariate risk factors for HSV-2 seropositivity included older age, female gender, and geographical place of residence. CONCLUSION: This large survey found a notably high seroprevalence of HSV-1, even among young female adolescents 15-19 years of age (80%). HSV-2 seropositivity was under 12% in all age groups surveyed in Poland, tending to be among the lowest overall HSV-2 seropositivity rates reported thus far in Europe.

Relationship between CMV infection and coronary heart disease.

Conflicting evidence implicating CMV infection in coronary heart disease (CHD) exists. In this work using serological methods (IgM-CMV by Western blot and IgG-CMV by ELISA) correlation between CMV infection and CHD was not found. On the other hand presence of CMV DNA in atherosclerotic plaques with absence in unchanged vessel indicates possible role of CMV infection in progression of this process.

[Influence of Chlamydia pneumoniae and cytomegalovirus infections on prevalence and the course of coronary artery disease]. / Wplyw zakazenia Chlamydia pneumoniae i wirusem cytomegalii na wystepowanie oraz przebieg choroby wiencowej.

Chlamydia pneumoniae (C. pneumoniae) as well as cytomegalovirus (CMV) are common pathogens found in about 50% of healthy western population. Many studies suggest a role of C. pneumoniae in development of coronary artery disease (CAD). CMV infection is also considered to increase risk of developing of CAD as well as restenosis after percutaneous coronary revascularization (PCI). The aim of our study was to evaluate a possible role of C. pneumoniae and CMV infections in both CAD development and course in patients (pts) undergoing PTCA. We enrolled 105 pts (mean age 56.4 years, 83 males) with angiographically documented CAD. Control group consisted of 63 healthy controls (mean age 47.25 years; 31 males). The study subjects were evaluated for presence of C. pneumoniae specific IgG antibodies (MIF test--MRL Diagnostic, USA; seroprevalence assumed when titre > or = 1/8). In 58 random PCI pts CMV specific IgG antibodies (ELISA Eti-Cytok-G PLUS--Dia Sorin) were evaluated. Pts were sampled at the time of PTCA. All PCI pts were assessed by angina questionnaire 5.9 +/- 2.6 months (mo) after the procedure with respect to clinical restenosis. C. pneumoniae IgG antibodies were detected in 37.1% of pts and in 22% of healthy controls (p < 0.05). After logistic regression was applied trend towards more frequent occurrence of C. pneumoniae specific IgG in CAD pts was shown (p = 0.10 OR = 2.4; 95% CI: 0.8-6.8). No significant correlation was found between anti-C. pneumoniae IgG presence or anti-CMV IgG titre and coronary atherosclerosis advancement. There was no significant difference in anti-CMV IgG titre between 9 pts who developed clinical restenosis 5.9 +/- 2.6 mo after PCI and the remaining pts. Our study results suggest a possible significant correlation between C. pneumoniae with CAD prevalence. We did not find a positive association of either infection markers with coronary atherosclerosis advancement. We did not find correlation of clinical restenosis after PCI with markers of CMV infection.

Flow cytometric enumeration of micronucleated reticulocytes: high transferability among 14 laboratories.

This laboratory previously described a single-laser flow cytometric method, which effectively resolves micronucleated erythrocyte populations in rodent peripheral blood samples. Even so, the rarity and variable size of micronuclei make it difficult to configure instrument settings consistently and define analysis regions rationally to enumerate the cell populations of interest. Murine erythrocytes from animals infected with the malaria parasite Plasmodium berghei contain a high prevalence of erythrocytes with a uniform DNA content. This biological model for micronucleated erythrocytes offers a means by which the micronucleus analysis regions can be rationally defined, and a means for controlling interexperimental variation. The experiments described herein were performed to extend these studies by testing whether malaria-infected erythrocytes could also be used to enhance the transferability of the method, as well as control intra- and interlaboratory variation. For these studies, blood samples from mice infected with malaria, or treated with vehicle or the clastogen methyl methanesulfonate, were fixed and shipped to collaborating laboratories for analysis. After configuring instrumentation parameters and guiding the position of analysis regions with the malaria-infected blood samples, micronucleated reticulocyte frequencies were measured (20,000 reticulocytes per sample). To evaluate both intra- and interlaboratory variation, five replicates were analyzed per day, and these analyses were repeated on up to five separate days. The data of 14 laboratories presented herein indicate that transferability of this flow cytometric technique is high when instrumentation is guided by the biological standard Plasmodium berghei.

Comparison of different protocols of MRC5 cell line permeabilization for detection of CMV IE antigens by flow cytometry.

The usefulness of four different protocols of permeabilizing human fibroblast cells for detection of CMV immediate-early antigens by flow cytometry was investigated. Permeabilizing protocols employed methanol, methanol/acetone, paraformaldehyde/Triton and FACSPermeabilizing Solution (Becton Dickinson). The best parameters (separation between positive stained CMV infected cells and negative control and detection of infected cells) were achieved for paraformaldehyde/Triton and FACSPermeabilizing Solution (Becton Dickinson).

Usefulness of hybridization and PCR methods in monitoring of CMV infection in renal transplant recipients.

The purpose of this work was to compare hybridization and PCR methods as diagnostic tests in diagnosing and monitoring CMV infection. The investigation was performed in a group of 24 renal transplant recipients treated with ATG. The results we obtained suggest that quantitative variant of hybridization is more useful in diagnosing the infection than PCR, because it enables to monitor the infection. DNA CMV level of about 60 pg/ml or the increasing level in the subsequent samples should be a sign to start antiviral therapy.

[Evaluation of the usefulness of various PCR method variations and nucleic acid hybridization for CMV infection in immunosuppressed patients]. / Ocena przydatnosci róznych wariantów metody PCR oraz metody hybrydyzacji kwasów nukleinowych w wykrywaniu zakazenia CMV u pacjentów z immunosupresja.

In diagnosis of CMV infection various laboratory methods are used. The methods based on detection of viral nucleic acids have been introduced routinely in many laboratories. The aim of this study was to compare nucleic acid hybridisation method and various variants of PCR methods with respect to their ability to detect CMV DNA. The studied material comprised 60 blood samples from 19 patients including 13 renal transplant recipients and 6 with acute leukaemia. The samples were subjected to hybridisation (Murex Hybrid Capture System CMV DNA) and PCR carried out in 3 variants: with one pair of primers (single PCR), nested PCR and Digene SHARP System with detection of PCR product using a genetic probe in ELISA system. The sensitivity of the variants ranged from 10(0) particles of viral DNA in nested PCR to 10(2) in single PCR. The producer claimed the sensitivity of the hybridisation test to be 3 x 10(5) and it seems to be sufficient for detection of CMV infection. The obtained results show that sensitivity of hybridisation was comparable to that of single PCR and the possibility of obtaining quantitative results makes it superior, on efficacy of antiviral therapy, especially in monitoring CMV infection in immunossuppressed patients and in following the efficacy of antiviral treatment.

Quantitative detection of CMV DNA by PCR and hybridizaton methods in renal transplant recipients.

The comparison of two quantitative tests: hybridization (Murex Hybrid Capture System) and PCR (COBAS AMPLICOR CMV Monitor) detected CMV DNA was made. Investigation of viral load in serum by PCR gave better correlation with clinical manifestation in renal transplant recipients.

Identification of cytomegalovirus (CMV) infection by different laboratory methods in renal transplant recipients undergoing triple-drug immunosupressive treatment.

Early diagnosis of CMV infection is very important mainly in transplant recipients because CMV infection is a frequent complication after transplantation. In this work we compared different laboratory methods: ELISA (IgG, IgM), Western blot,shell vial, antigenemia assay (pp65), the immunofluorescent method with epithelial cells from urine (IF), DNA in leukocytes by PCR and DNA in leukocytes by hybridization (HCS) to estimate the most proper method for diagnosis of CMV in renal transplant recipients. This preliminary study showed that HCS, PCR and Western blot are sensitive methods for detecting CMV infection. Using HCS in quantitative variant we obtained a very good correlation between DNA load and clinical symptoms.

[Diagnosis of CMV infections in patients diagnosed with acute leukemia]. / Diagnostyka zakazen CMV u pacjentów z rozpoznaniem ostrej bialaczki.

The purpose of this paper was to estimate the best method of CMV diagnosis in leukemic patients. Materials from 9 patients (serum, heparinized blood and urine) were investigated by serological methods (ELISA and Western blot), for the presence of specific antigens and virus capable of replication, and also by genetic methods (hybridization and PCR). It seems, that the best method for CMV diagnosis in leukemic patients is hybridization performed in quantitation variant, and that DNA CMV level of approximately 20 pg/ml of blood was not linked to symptomatic infection.

["Shell vial" method in diagnosis of cytomegalovirus infections]. / Metoda "shell vial" w diagnostyce zakazen wywolanych wirusem cytomegalii.

The paper presents description of "shell vial" method and results obtained in the study of 331 materials derived from 3 different groups of patients. Problems of referring materials to laboratory, choice of type of material and method of staining are discussed. Also advantages (detection of virus capable of replication) and disadvantages (low sensitivity, toxicity of materials for detector cells) are discussed.

Characteristic of humoral response of elderly to inactivated influenza vaccine.

Eighty persons immunized with inactivated influenza vaccine in 1990-1991 epidemic season were examined. Serological tests: hemagglutination inhibition (HI), lectin neuraminidase (LN) and Western blot were performed with serum specimens taken before and 3-4 weeks after vaccination. Distribution of anti-influenza antibodies in the subclass was also examined. The results revealed humoral response to vaccine strains of influenza virus and also to strains caused infections in the past.

[The use of dot blot method for the estimation of IgG antibodies among persons vaccinated against influenza]. / Zastosowanie metody dot blot do oznaczania przeciwcial klasy IgG u ludzi szczepionych przeciwko grypie.

The level of IgG antibodies before and post influenza vaccination for 6 strains of influenza A virus (4 representative for pandemics and 2 vaccine strains) was tested by dot blot method. Examinations was performed with 4 age groups (20 persons each), selected on the base of strain which probably caused primary infection in given group. No differences in answer to particular influenza A virus strains consistent with "original antigenic sin" were observed. High sensitivity of dot blot method was confirmed.