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Cytomegaloviruses are common worldwide, with variable frequency of infections. The infection in pregnancy may lead to pregnancy loss or serious sequelae for the child. To understand the risk posed by CMV in Poland we conducted cross-sectional study on women aged 15–49 basing on existing serum bank. Age dependent CMV incidence, the rates of congenital infection and sequelae were modelled from sero-prevalence, literature and demographic data. The overall anti-CMV IgG prevalence was 81.9% increasing from 74.3% in < 30 years old to 94.3% in subjects 45+ years old. The lowest incidence was estimated at the age of 15 and the highest at the age 34 (3.8 and 8.95 respec- tively/100 women/year). The estimated rate of cCMV varies from 22.4 to 37.2 per 1000 live birth depending on the assumptions made. The proportion of cases due to secondary infection ranged from 34.8% to 49.9% accordingly.

Hepatitis B and C viruses have the potential for generating autoimmune processes with dysregulation of the cytokines, Th1 chemokines and molecular or epitope mimicry between self-antigens and viral proteins. These phenomena include the production of autoantibodies, but their clinical significance is still unknown. The aim of this study was to determine the presence of autoantibodies in HCV compared to HBV infected patients, and to find the association of their presence with the age, sex and microscopical changes in the liver. The incidence of autoantibodies presence parallel to the liver biopsy was assessed in 363 consecutive patients with hepatitis C, and 71 patients with hepatitis B. It was found that 66 (18%) HCV and 8 (11%) HBV infected patients had positive results. Pathomorphological changes typical for autoimmune hepatitis were not found in this group of patients with viral infection. The HCV infected patients with autoantibodies were significantly older than patients without autoantibodies. This correlation could be connected with the worsening of the self-mechanisms protecting against autoimmune reactions and the longer duration especially of HCV infection. No such similar correlation was observed in the group of HBV infected patients.

Epstein-Barr virus (EBV) establishes latency in the resting memory B-cell compartment. It has been recently suggested that maintenance of chronic infection is dependent on periodic reactivation. Although the stimuli for EBV reactivation in vivo during natural infections are largely unknown, there is evidence indicating that heterologous infections could trigger herpesviruses reactivation. The purpose of this work was to identify the influence of Toll-like receptors stimulation on EBV replication in EBV latently infected Burkitt lymphoma cells (P3HR-1, Raji and Namalwa). The cells were stimulated with Pam3CSK4 (synthetic triacylated lipoprotein), PolyI:C (synthetic analog of dsRNA), LPS (lipopolysaccharide from E.coli), measles virus (MeV) and PMA (phorbol myristate acetate). Non-stimulated cells (NS) served as control. EBV expression was investigated at mRNA level for three viral lytic genes: BZLF1 (immediate early, ZEBRA), BALF2 (early, EA) and BcLF1 (late, VCA). Additionally, the effect of stimulation on NF-kBp65 and inflammatory cytokines (IL-lb, IL-6, IL-8, IL-10, IL-12p70, and TNF) was investigated. Stimulation of TLRs led to limited changes in EBV expression manifesting as increase of ZEBRA at mRNA level in cells treated with PolyI:C and Pam3CSK4. Stimulation with PolyI:C, Pam3CSK4 and LPS also lead to considerable increase of NF-kBp65, while increased levels of inflammatory cytokines were observed for IL-8, TNF and IL-6 in cells treated with PMA and MeV. In conclusion, the results of our experiments support the suggestion that TLRs stimulation with microbial ligands influences EBV virus replication.

Toll-like receptors (TLRs) are an important component of a innate immune system. Stimulation of TLRs, through action with helper T cells, could change Th1/Th2 balance and thus affect adaptive immune response. Receptors TLR2 and TLR4 play important role in immune response to measles virus. The aim of this work was stimulation of the peripheral blood mononuclear cells (PBMC) and determination of Toll-like gene expression in these cells.PBMCs from 20 healthy donors were stimulated with measles viruses and ligands for TLR2 and TLR4. For examinations the real time RT-PCR (QuantiFast® Assay, Qiagen) was used. The expression of Toll-like receptors was determined on muRNA level, using real-time one step RT-PCR (QuantiFast® Assay, Qiagen) with simultaneous detection of TLR2 and TLR4 genes and housekeeping gene (GAPDH).Virus-specific influence of wild measles virus strains activity on PBMC derived from vaccinated seronegative individuals manifested in higher level of expression of TLR2 and TLR4 genes in compare to the expression of these genes in PBMC of seropositive individuals.Toll-like receptors participate in the development of immune response to measles virus.

In Poland, large-scale serologic surveys carried out in 1965–1972 revealed regions of Poland with particularly high prevalences of antibodies against tick-borne encephalitis virus (TBEV). The information provided by the routine surveillance of communicable diseases during 1970–2005 indicated, however, that the geographic distribution of the disease is limited to a few eastern and south-western regions of Poland (defined as endemic for the purpose of this study). In the present serologic survey, 1498 human serum samples collected in 1996–2005 were randomly selected from a serum bank, and 358 goat serum samples were collected from milk-producing farms in selected areas of Poland 2002–2006. Thirty-nine human samples were positive for anti-TBEV antibodies, with an overall seroprevalence of 2.6%. Seroprevalence in endemic provinces ranged from 0.8% to 4.3%, and seroprevalence in non-endemic provinces ranged from 1.9% to 4.3%. In endemic, compared to the non-endemic provinces, the highest seroprevalence was found in the age group >60 years (7% vs. 1%) and in inhabitants of villages (3.9% vs. 1.8%). In non-endemic, compared to endemic provinces, the highest seroprevalence was detected in the age groups 30–39 years (5% vs. 2%) and 40–49 (4% vs. 0%), and in inhabitants of large towns inhabited by >100,000 people (4.1% vs. 2.5%). Out of 358 goat samples, 17 (4.7%) were positive for anti-TBEV antibodies. Seroprevalence in goats reached 14/151 (9.3%) in endemic, and 3/207 (1.4%) in non-endemic provinces. The present study indicates the possible existence of endemic foci in north-western provinces of Poland, in which barely any cases were reported during 1970–2005. The socio-demographic profile of seropositive subjects in non-endemic regions suggests that they might have been exposed to TBEV during travels to known endemic regions. This would mean, however, that they were missed by the surveillance system. A thorough review of diagnostic protocols in non-endemic regions and work towards an improvement of the TBE surveillance sensitivity is necessary.

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.

Current studies have indicated many environmental factors, such as pesticides, that cause immune system disorders through inducing changes in humoral and cellular responses thereby increasing the risk of contracting infectious diseases and cancer. The literature suggests that low exposures to certain organophosphorus pesticides stimulate the immune system, whilst high exposures result in decreased function. Precise mechanisms for the fall in immunocompetence are often unclear, however it can be predicted that the intimate interaction between the nervous and immune systems can potentially lead to toxicity.To determine the effects of organophosphorus pesticide, chlorpyrifos that is often used in Poland, on selected immunological responses, such as immune-competent cell proportions formed experimentally in-vivo by cells of Wistar rats during subchronic exposures after 45 and 90 days.The test was carried out on ten male and ten female Wistar rats in each of three test groups, who received 3 chlorpyrifos doses for 90 days intragastrically, according to OECD guidelines (No. 401). Two control groups were given olive oil. After completion, the animals were deeply anaesthetised by a mixture of ketamine (Vetaketam) and xylazine (Vetaxym). Immuno-competent cells were profiled by a commercial monoclonal antibody method. In order to measure the dynamics of any changes, the aforementioned immunological responses were investigated after 45 days using the same procedures for obtaining the relevant biological test material.Test animals exposed to chlorpyrifos had altered number of white bood cells which were either increased or decreased relative to controls after 45 and 90 days for all exposure levels used.The study demonstrated changes in white-blood cell (lymphocyte) response profiles, reflecting an immunomodulation although such changes were equivocal, where both suppression and stimulation were observed.immunomodulation, immune system, lymphocyte, organophosphorus pesticides, chlorpyrifos.

The paper discusses the role of anti-measles antibodies for protection and significance for epidemiological studies determination of antibodies by different serological methods. The comparison of anti-measles virus antibodies levels measured by enzyme immunoassay (EIA) and Plaque Reduction Neutralization Test (PRNT) was described. It was found that the 200 mIU/ml of anti-measles activity measured by PRNT (level protection against symp- tomatic disease) is equivalent of 636 mIU/ml measured by EIA (Enzygnost®Anti-Measles Virus/IgG, Simens).

Cytomegalovirus infection (CMV) is one of the most common viral infections during pregnancy and one of the most common causes of birth defects in newborns. CMV infection occurs mostly through close contact with small children who can secrete the virus in saliva and urine. Children, especially in preschool and early school can also be a source of infection with other herpesviruses. The aim of the study was to determine the prevalence of active infections caused by viruses from the family Herpesviridae (CMV, EBV, VZV) among members of families with children.The study included 24 families raising children aged from 2 to 18 years. From all members of the families (46 adults and 39 children) saliva samples were collected from which DNA was extracted. The isolated DNA samples were tested for the presence of CMV, EBV, VZV genetic material by nested PCR. In addition, each family carried out a survey.The presence of CMV DNA in saliva samples were detected in members of 7 families and the presence of EBV DNA were detected in members of 11 families. Total DNA of CMV was detected in 8/85 samples of saliva (9.41%), of which 1/46 adults (2.17%) and 7/39 children (17.95%) and EBV DNA was detected in 18/85 tested saliva samples (21,18%) - 13/46 samples from adults (28,26%) and 5/39 samples from children (12,82%). VZV DNA was not detected in any of the tested saliva samples. The obtained results indicate that the active, asymptomatic infections with lymphotropic herpesviruses are common and affect more than 10% (CMV) and 20% (EBV) subjects.

Obtaining a reliable laboratory test result depends on many factors, among which preanalytical factors play a significant role. The aim ofthis study was to determine the effect of temperature, storage time of samples and number of freezing and thawing cycles on the results of tests carried out by PCR.The study was conducted in a model of cytomegalovirus (DNA) and with four types of clinical material: whole blood, serum, cerebrospinal fluid, urine and archival samples. After contamination with CMV clinical samples were incubated under various conditions for a specified period of time or subjected to cycles of freezing and thawing. The presence of DNA - CMV were determined by nested PCR method.The results revealed that in the case of applied levels of contamination and in model used in the study: 1) CMV DNA in samples of clinical material (blood, serum, cerebrospinal fluid, urine) for testing by PCR remains stable for at least 5 days at room temperature or refrigerated and at least 7 days when frozen, 2) Repeated freezing and thawing clinical specimens has no influence on the result of the presence of viral DNA, 3) Isolated viral DNA is stable in the temperature of the freezer and can be subjected to at least 20 freeze-thaw cycles without affecting the PCR results.Viral DNA in clinical specimens examined in the applied experimental conditions is stable.

Many environmental factors, including pesticides, cause immunological system disorders by inducing changes in humoral and cellular response. They may stimulate or trigger immunological autoaggression, hypersensitivity and allergy, as well as lead to immunosuppression, thus increasing the incidence of infectious diseases and cancers. Such activity is also attributed to organophosphorus compounds used in agriculture as insecticides, and commonly in households as biocides. The aim of the study was to define possible mechanisms of the immunotoxic activity of the chlorpyrifos (an organophosphorus compound) on experimental animals following their exposure to the compound via the oral route. The present study attempts to define the influence of chlorpyrifos on the profile of subpopulations of immunoactive cells: B, T, CD4+, CD8+, and NK, and on their phagocytic activity in an experimental in vivo model. For this purpose, the Wistar rats, were exposed orally to increasing doses of chlorpyrifos: 0.1 LD(50), 0.15 LD(50), 0.2 LD(50), 0.3 LD(50) and 0.4 LD(50) for 28 days. In the study animals, we failed to demonstrate a statistically significant decrease in the phagocytic activity of the granulocyte.

The increase of measles incidence in Poland was recently observed. Furthermore, the analysis of routine serological tests performed in the department of virology, NIPH-NIH revealed, that nearly half of young people (20-30 years old) have no antibodies against measles virus. The paper presents results of IgG specific for measles virus prevalence in the sera of vaccinated and unvaccinated subjects, which aimed to make the selection of groups for immunological memory research.Total of 100 persons were examined based on results of determination of the presence of IgG anti-MeV: 26 people born before and 74 born after 1972 year. From this group, 55 participants were selected for further study and divided into 3 groups (1) subjects born before 1972, unvaccinated against measles and seropositive as a result of natural infection, (2) subjects born after 1972, vaccinated against measles but seronegative or with traces ofanti-MeV IgG presence, (3) subjects born after 1972 seropositive due to vaccination. Selected persons are subject to further examinations include determination the number of leukocytes and lymphocytes profile.The level of anty-MeV IgG antibodies in subjects after natural infection was significantly higher compared to levels obtained by vaccinations. No significant differences in the immunological parameters which could influence on immune response were observed.Obtained results lead to search other factors that may affect the weak postvaccinal humoral response.

Obtaining a reliable laboratory test result depends on many factors, among which preanalitical factors play a significant role. The aim of this study was to determine the effect of temperature, storage time of samples and number of freezing and thawing cycles on the results of tests carried out by RT-PCR. The study was conducted in a model of measles virus (RNA). The results revealed that: (1) Samples of clinical material (serum, cerebrospinal fluid, urine) for testing by RT-PCR can be stored for at least 5 days at room temperature or refrigerated and at least 7 days when frozen, (2) Freezing and thawing three times, samples of clinical material does not affect the outcome of the presence of viral RNA, (3) Isolated viral RNA is stable at freezer temperature and can be subjected to at least 20 freeze-thaw cycles without affecting the results.

Elimination of measles is one of the priority plans of WHO. The success of this plan depends on the development of long lasting, postvaccinal immune response. The aim of this study was to present the effect of stimulation with different strains of measles virus on the expression of T-helper cell (CD4+ T) early activation markers in people with different history of measles infection and to determine the correlation between the activation and dose of virus used for stimulation. The study was conducted using material derived from two patients: one seropositive due to natural infection and one vaccinated, with traces of anti-MeV IgG antibodies. In the CD4 T helper cells, the expression of CD69 receptor and the ability of the cells to produce INF after stimulation with the vaccine-derived or wild-type strain of measles virus was determined. For antigen-specific stimulation the virus suspension containing about 100 infectious particle, its tenfold and hundredfold dilutions was used. We found that the expression of T-helper cells early activation markers depended on the strain of the measles virus used for the stimulation, type of the immune response (postvaccinal, natural infection), and in the case of CD69 expression also on the dose of the virus used for the stimulation.

To achieve measles elimination, an efficient surveillance system for rash illnesses is necessary. The aim of the present study was to ascertain which viruses, other than measles, are causing measles-like illnesses (MLIs) in Poland. Serum samples (n=278) collected from MLI cases and submitted to the National Reference Laboratory during 2006-2007 were investigated for anti-measles (MeV), rubella (RUBV), parvovirus B19 (B19V), Epstein-Barr (EBV) and herpesvirus type-6 (HHV-6) IgM presence. Age was strongly associated with MLIs etiology. In the youngest age group, 0-4 years, MeV and HHV-6 infection were prevailing, while in group of 5-9 years--RUBV and B 19V. Measles was confirmed more often in patients with high fever (p0.001) and with rash lasting longer than 5 days (p0.001). The type of rash was not significantly associated with MeV infection. Our results strongly suggest that according to WHO EURO strategic plan, Poland is close to elimination phase. High number of MLIs were caused by pathogens other than measles. Addition of anti-B 19V IgM testing to routine MLIs screening protocol may improve system performance in the more advanced stages of measles elimination.

The purpose of this study was evaluating influence of repeated cycles of thawing and freezing serum samples on IgG and IgM stability. Serum positive samples for anti-cytomegalovirus and antimeasles virus IgG and/or IgM were aliquoted. Tubes containing 100 microl aliquot were frozen at -20 degrees C. Samples were repetitively thawed and froze in one to ten cycles. Antibodies presence was examined with commercially available ELISA tests. All samples reminded positive even after ten repeated thaw/freeze cycles.

In Poland, 200 cases of tick borne encephalitis are annually registered. It is about half of all registered cases of viral encephalitis. Immunity acquired after vaccination with TBE inactivated vaccine persists defined time dependent on number of dose of vaccine and individual features of vaccinated person. The aim of this paper was to determine the frequency of IgG antibodies against TBE virus among forestry workers from Białowieza National Park with regard to information about vaccination against TBE virus. In the group of 59 persons vaccinated against TBE virus, 54 (91.5%) had IgG antibodies against TBE virus. In the group of 32 persons (non-vaccinated or for whom the information about vaccination was not given) 26 (81.3%) were seropositive. Relationships between VE value (determining level of antibodies), age and period passed from the last dose of vaccination were analysed. Taken to account the most disadvantageous circumstances (age = 60, regression intercept minus two standard deviation) antibodies level of VE = 11 (lower limit of positive result) remains for 2,72 years from last dose of vaccine against TBE virus. This result confirm recommendation about necessity to use of complementary dose within 3-5 years, at the same time it should be considered recommendation of necessity ofrevaccination persons 60 years old or above as frequently as every 3 years.

The aim of this study was to analyze histopathological changes in the liver and serum inflammatory cytokine level in hepatitis C virus (HCV)-infected patients with and without cryoglobulinemia. The study group consisted of 34 patients with chronic hepatitis C, confirmed by serological and virological markers. Ten out of 34 patients had cryoglobulinemia. The control group consisted of 21 healthy persons. Liver biopsy specimens of HCV-infected patients were evaluated by light microscopy using the grade and the stage according to Batts and Ludwig classification. The quantitative measurements of IL-1β, IL-6, IL-8, IL-10, IL-12p70, and tumor necrosis factor in sera were performed by flow cytometry. The mean age of HCV-infected patients with cryoglobulinemia was higher than age of HCV-infected patients without cryoglobulinemia. Microscopic examination of liver biopsy specimens revealed necroinflammatory activity slightly more prominent in patients with cryoglobulinemia. The most prominent inflammatory changes connected with abundant lymphoid aggregates in most of the examined portal tracts and piecemeal necroses were diagnosed in patients with several extrahepatic manifestations, such as cutaneous manifestations, nephrotic syndrome, polyneuropathy, and arthropathy. Liver fibrosis was similar in patients with and without cryoglobulinemia. The serum levels of all proinflammatory cytokines, especially IL-8, were significantly higher in the patients with cryoglobulinemia in comparison with the patients without cryoglobulinemia and healthy persons. All microscopic features did not correlate with the level of any investigated proinflammatory cytokines.

A varicella zoster virus (VZV) is the first herpesvirus for which a vaccine was developed. Since 1999, the varicella vaccine is licensed in Poland and recommended for use in adults without history of a varicella infection, and in children and young adults with remission of acute leukemia. While serological data is essential to assess the appropriate vaccination programme, we conducted the first in Poland serosurvey on a representative group of Polish population aged 1-19. Serum samples were selected from a serum bank collected in 1995-2004 with a catchment area of the all geographical regions of Poland. A total of 1300 serum samples collected over 9 years (1995-1996, 1998-2004) were selected using a stratified sampling design (stratification by age). Samples were selected, consisting of 100 samples for each 1-year band of age groups 0-9 years, and 40 samples for each 1-year band of age groups 10-19 years. IgG serum antibodies specific to VZV were detected using an indirect enzyme immunoassay and the antibody level was expressed in international units per millilitre (mIU/ml) and was refered to the international standard for VZV immunoglobulin of 50 IU. The overall seroprevalence estimate, adjusted for sampling design for the age group 1-19 was 76.6% (95% CI: 74.6%-78.7%). Seroprevalence correlated closely with age (p0.0001) and among 18 and 19 year olds reached 95% and 98% respectively. No association was found between gender, rural/urban areas and geographical regions of Poland. For samples collected over the 5 year period (2000-2004), evidence of overall differences in seropositivity over these years was not observed. In Poland VZV vaccination is provided only for a limited group of high risk patients. The possible updates in the immunization program are discussed and the results of the presented study can contribute valuable information to base the vaccination policy decisions.

The aim of presented paper was to compare the results of EBV tests performed using real-time PCR (LightCycler EBV Quant Kit--Roche) and "home made" nested PCR in patients with serological confirmed actual or recent EBV infection. In serum samples the presence of IgM and IgG antibody to virus capsid antigen (VCA), IgG antibody to nuclear antigen (EBNA) and early antigen (EA) was tested. Serum samples with the presence anty-VCA IgM (I Group) or with anty EA IgG and without anty-VCA IgM (II Group) were tested by PCR. Real-time PCR results were compared to nested PCR. In both PCR methods 57/60 (95%) concordant results were obtained (10 positive and 47 negative samples). In two serum samples positive result was obtained only by real-time PCR, in one serum sample only by nested PCR. EBV DNA was detected only in serum samples from I Group. Negative EBV DNA detection may be correlated to phase of infection (time after onset of disease) in which the sample was collected, as well as with fact how long EBV is presence in blood. The results indicate that real-time PCR is a reliable tool for diagnosis of primary EBV infection early in the course of disease and may serve as test of confirmation in serologically indeterminate EBV infection (doubtful cases).

The aim of this study was to perform the validation of nested RT-PCR method for detection of measles virus genome in laboratory of Virology Department (NIZP-PZH) conditions. The PCR reactions were made with specific primers for nucleoprotein gene. The validation of nested RT-PCR assay includes determination of limit of detection (LOD), specificity, intra- and inter-assay precision and influence of materials from clinical samples on the PCR reaction. The detection limit of nested RT-PCR was 255-510 RNA MeV/ml. The obtained results reveal high precision and lack of substances present in clinical samples influence on the PCR reaction. The application of nested RT-PCR assay for viral genomes other than MeV gave negative results. The obtained results indicated that the nested RT-PCR fulfilled validation criteria and can be used for routine laboratory diagnostics of measles virus.

The most of registered in Poland cases of encephalitis and meningitis have viral aetiology. Confirmation of viral central nervous system (CNS) infection and diagnosis of pathogenic agent is critical for therapeutic treatment, especially if antiviral chemotherapy is available. The aim of this work was analysis of routine laboratory results obtained in Laboratory of Department of Virology NIZP-PZH by examination of materials obtained from 82 medical canters in Poland in aim of CNS infection confirmation. Materials, cerebrospinal fluid (CSN) n=277, and CSN together with serum (n=452) were obtained from patients aged from 3 days to 83 years. Accordingly with the range of tests performed in Laboratory of Department of Virology NIZP-PZH, obtained samples were examinated for 11 viral infections: HSV, CMV, EBV, VZV, HHV-6, HHV-7, TBE, measles, mumps, rubella and enteroviruses. Confirmation of viral infection was obtained in 104 out of 729 tested patients (14.3%). The highest number of confirmations was obtained in case of TBE infection (18.4%) and HSV (9.2%). The methods gave the highest number of confirmations were testing of intrathecal IgG synthesis (14.4%) and presence of IgM in serum (10.3%). If test was conducted only with CSF, confirmation of viral infection was obtained in 13 cases (4.7%). In conclusions it was ascertained that testing CSF and serum samples together greatly increase possibility of etiological agent detection and a range of ordered tests (i.e. intrathecal synthesis versus PCR) should account dynamics of pathological process.

Cytomegalovirus infection is one of the main problem in immunocompromised patiens. Quantitative assessment of CMV load (viral load), rate of increase of load and determination of DNA level above which the likelihood of disease is high (viral load thresholdfor disease) have significant prognostic and therapeutic importance at transplant recipients. The aim of this work was the comparison of 3 quantitative molecular techniques and assessment the threshold for disease for each of them. The study was undertaken with 37 samples of serum and the whole from 17 renal transplant recipients. Part of samples (n=16) comes from symptomatic patients, and were taken in period of clinical symptoms demonstration. The samples ware investigated by hybridization method (HC) performed accordingly to Hybrid the Capture procedure, (r-t PCR) Amplicor test (COBAS AMPLICOR CMV the Monitor test) nd real time PCR (r-t PCR). In 21 out of 37 samples DNA CMV was detected by all 3 methods, 2 samples gave concordant negative results. The CMV DNA level measured by all 3 methods was significantly higher (p0.05; t-Student test) in samples from symptomatic patients than from asymptomatic: 4.79 versus 3.58 for HC; 3.06 versus 1.36 for PCR-Amplicor and 4.23 versus 2.88 log DNA copies/ml for r-t PCR. The threshold for disease connected with high likelihood of disease (p0.05; Fisher test) was established at 4 log for r-t PCR method, 4,61 for hybridization and 3 log DNA CMV copies/ml for PCR Amplicor.

Epstein-Barr virus (EBV) infection is common all over the world. Primary EBV infection can lead to latent or chronic infection resulting in lymphoproliferative diseases.of this study was to evaluate the effectiveness of laboratory methods used to detect EBV and to monitor EBV infections in children with acute leukaemia.we conducted the study on 30 children with acute leukaemia. For the present analysis we included 8 patients from whom samples were also taken three times. Studies were made on serum, blood and bone marrow. Specific IgM and IgG antibodies for viral capsid antigen (VCA), nuclear (EBN) and early antigen (EA) were tested by ELSA. The presence of DNA virus was estimated by nested PCR. The control group were 11 subjects, without chronic or neoplastic disease, undergoing routine laboratory tests in the Virology Department.on admission serologied tests indicated past EBV infection in 6 out of the 8 children. Primary infection was detected in 2 patients, in one of them on day 1 of the observation and on day 22 in the other. DNA EBV was found by nested PCR in only one of the 8 infected children.1. Results of serological investigations indicated the type of EBV infection in our patients. 2. Confirmation of EBV virus active replication by nased PCR was obtained in only 1 out of 8 patients. 3. In order to assess the effectiveness of serological and molecular methods in evaluation of EBV infection type in children with acute leukaemia, it is necessary to investigate a larger group of patients and taken at least three times.

Laboratory confirmation of EBV infection requires proper methods and schema of investigation adequate to aim of diagnostic procedure. In paper the results of routine diagnostic tests of EBV infection performed in Department of Virology NIH in 2005-2006 years was included and also, evaluation of usefulness of different laboratory methods was done. Based on results of ELISA tests 10,7% routine investigated subjects was classified as primary EBV infection, 20,1% was seronegative, 7,4% was classified as reactivation of latent infection and serological markers in 45,6% subjects pointed past EBV infection. Positive result of PCR method was obtain in 11,2% samples subjected of routine laboratory investigation. Comparison of specific and non-specific serological methods results (ELISA versus tests of heterophile antibodies) showed the high percentage of false negative results in children tested by non-specific tests. PCR results in serum samples from patients with primary infection (confirmed by serological tests) were positive in 15% cases only. Based on analyzed results it could be stated that reliable confirmation of infectious mononucleosis, as primary EBV infection, is detection of specific IgM antibodies and in case of heterophile antibodies tests the possibility of false negative results, mainly in children, must be taken into account. The most proper samples for PCR method are whole blood, sections of tissue or cells from swabs.

A spread of hantavirus infections causing hemorrhagic fever with renal syndrome is one of the rising epidemiological problems in the world. There is evidence that this type of viruses exists in almost all European countries. It is known that in Poland all the species of wild rodents hosting European hantaviruses are present. However, the data on hantavirus are very limited and information on incidence and prevalence of hantavirus infections among humans and animals in our country is lacking. Up to now, only one study in Poland has dealt with hantavirus infections in humans. Therefore the aim of the presented study was to show whether those infections are present in Poland and how prevalent they are. Two risk groups of hantavirus infection were investigated: group I--people with renal disorders (hospitalized patients with proteinuria and people suspected to hantavirus infection; n=30 people) and group II--healthy people employed at the universities, scientific institutions who due to their profession have contact with small rodents as well as students who captured those rodents when practicing or preparing their master theses (n=76) and forest workers (n=86). No one from the group of patients with renal disorders was positive for hantavirus Puumala IgG. Moreover, none of the forest workers had evidence of hantavirus infection. The hantavirus IgG were found only in the group of mammologists--19% persons were positive. Mammologists are an occupational subpopulation who remain in direct contact with host and/or with host excretions (virus is present in urine, feces, saliva of infected animals) and seem to be the most sensitive indicator of hantaviruses presence in environment. These results prove occurrence of hantavirus infections in Poland, however it seems that they are not very prevalent.

Serological diagnostic tests were introduced for detection of immunoresponse against to etiological agents of current infectious diseases. Interpretation criteria for those tests should eliminate unspecific reactions, cross-reactivity with related viruses as well as traces amounts of immunological response against to the past infections. In the case of past hantaviruses infections for obtaining an adequate seroepidemiological data revalidation of this tests are required. In present studies Puumala IgG results obtained in sera from healthy population had been analysed by statistical method and interpretation criteria were recalculated. Panels of 86 sera from forest workers and 47 zoologists working with small mammals were evaluated for hantavirus specific IgG (Hantavirus Puumala ELISA test). Puumala specific antibodies were detected in 7 zoologist's sera and in 5 sera gave equivocal results. All sera collected from forest's workers were negative. Statistical analysis based on negative results in forest workers group suggests that the cut off of the ELISA test suitable for diagnosis of suspected nephropathia epidemica cases is too restrictive for seroepidemiological research of hantavirus in healthy population.

The aim of the study was to assess the seroprevalence of parvovirus B19 in the Polish general population. Serum samples (n=2,500) were randomly selected from a serum bank comprising materials coming from inhabitants of all 16 polish provinces, collected by the Department of Virology in 1995-2004. Testing was performed using ELISA kits. Association of seropositive results with age, gender, residence type, and geographical location was assessed by bivariate and multivariate analyses. Of the 2,500 samples tested, 1,321 (52.9%) were seropositive for parvovirus B19. First peak of seropositive persons was observed in pre-school children, the proportion of seropositives achieving 80% among 40 year-olds. Up to 40% women of child-bearing age were established susceptible to parvoviral infection. No association between seropositivity and gender, year of sample collection, residence type and place was observed. Analysis of proportion of seropositive children aged 1, 2 and 3 by year of sample collection permitted identification of epidemic year occurrence in 1999. Results of the present study indicate that parvovirus B19 infections are widespread in Poland and are not associated with gender, residence type or geographic region. Up to 40% women of child-bearing age were established susceptible to parvovirus B19 infection, which indicates that it can constitute an important factor of miscariages, especially during epidemic years.

CMV infection can undergoing as primary or secondary infection, which can be represented by reactivation of endogenous infection or by superinfection with another viral strain. Taking for consideration high prevalence of seropositive persons it may be concluded that superinfections occur quite often. For the other hand it is known, that differences among CMV strains are deep and regard as well viral structure as its biological properties. The aim of this study was investigation of immunological profile in range of cytokines production after stimulation with different CMV strains. The study was conducted with leukocytes obtained from CMV seropositive person cultivated and stimulated in vitro with AD169, Towne and Davis. After 24, 48, 72 and 96 hours incubation concentrations of IL-2, IL-4, IL-5, IL-10, INF-gamma and TNF-alpha in supernatants was measured by flow cytometry with Cytometric Bead Array technique. Obtained results allow to conclude that difference in level of stimulated cytokines and their panel is dependent on viral strain used. The experiment also allows to optimisation of stimulation conditions.

The interaction of CD28 with one of the B7 molecules (CD80 and CD86) on professional antigen-presenting cells (APC) is generally considered to be the most important co-stimulatory signal for T cell activation. Several lines of evidence suggest that dendritic cells (DC), the most potent antigen presenting cells known, play a role in the immunological control of herpes simplex virus (HSV) infections. The fact that CD86 is strongly up-regulated together with other co-stimulatory molecules during DC maturation suggests that it plays an important role in induction of immune response. To determine the effect of virulence on up-regulation of CD86, we stimulated population of spleen cells enriched in dendritic cells by HSV-1 strains characterised by different pathogenicity. We analysed cells, which express CD45 molecule. HSV-1 ts, earlier described as less virulent for mice, stimulated an increased expression of co-stimulatory molecule CD86 than wild strain did.

In this work evaluation of usefulness of monoclonal antibodies (MAbs) CCH2 and AAC10 directed against early--pUL44(DB52) and late--ppUL83(pp65) CMV antigens, utilized in Department of Virology, NIH for routine diagnosis of CMV infection by shell vial and pp65 antigenemia assay, for determination of CMV antigens by flow cytometry in human leucocytes, isolated, infected and cultivated in vitro was presented.

Vitamin deficiencies and infections caused by viruses and other pathogens often coexist and exhibit complex interactions. Malnutrition, vitamin A deficiency and infections among populations of the developing countries, these are the leading cause of death, particularly in children. Several micronutrients were in study, vitamin A deficiency is strongly involved in measles and diarrhea. There are also some scientific data against this thesis. These vitamins, virus infection and immune function interactions are briefly reviewed in this article.

Against many viral diseases caused for example by HSV, EBV, CMV, HIV, RSV, HCV for which vaccines are not available, chemiotherapeutics seem to have the principal significance. High progress in development of new antiviral compounds is observed. In addition to synthetic compounds a large number of naturally occurring substances have been shown to posses antiviral activity. One of such substance is tannic acid. In this study comparison of antiviral activity of tannic acid, acyclovir (ACV) and ganciclovir (GCV) against cytomegalovirus (CMV) is presented. The MRC5 cells infected with CMV and treated with different compounds were analyzed by flow cytometry and cythopatic effect inhibition test for inhibition of virus replication and by MTT assay for cytotoxity. It has been shown that tannic acid has antiviral activity against cytomegalovirus and that expression of virus antigens measured as median fluorescence intensity (MFI) by flow cytometry can be used for evaluation of virus replication.

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.

Background Tick-borne encephalitis (TBE) is found in limited endemic foci in Poland. Lack of diagnosis limits disease detection in non-endemic provinces. Methods In 2009, we enhanced TBE surveillance to confirm the location of endemic foci and inform vaccination policy. In 105 hospitals located in 11/16 provinces, we identified suspected TBE cases through admission ICD-10 codes indicating aseptic meningo-encephalitis or from specimens tested for TBE. The National Reference Laboratory confirmed cases at no cost, by testing serum and/or cerebrospinal fluid using ELISA method. We calculated TBE reported rates as the number of confirmed TBE cases per 100,000 inhabitants. Adjusting to neighbouring districts, we classified districts as non-endemic ( = 0.1 to = 1 to = 5). We compared surveillance data obtained in 2009 with 2004–2008 baseline data. Results Among 166,099 admissions, we identified 1,585 suspected TBE cases of which 256 were confirmed. Physicians reported more suspected cases among patients