Coxsackieviral infections involved in aseptic meningitis: a study in Slovakia from 2005 to 2009.

A wide range of diseases is associated with enteroviruses.They are reported to be responsible for viral meningitis, especially in children, but also in adults.This study analysed infection with eight selected coxsackievirus serotypes as the cause of aseptic meningitis in 480 patients in Slovakia from 2005 to 2009,using a quantitative assay for the detection of intrathecal antibodies. Intrathecal production of antibodies against selected coxsackieviruses was proved in 21%of these patients. A significant decrease from 35% in 2005 to 8,5% in 2009 (p=0.004) in the proportion of patients with proven intrathecal production of virus specific antibodies was observed during the study period. We conclude that coxsackievirus B4 was the endemic serotype in Slovakia and was responsible for most cases of coxsackieviral meningitis in the study period.

[Use of PCR in diagnosis of enteroviral infections]. / Pouzitie metódy PCR v diagnostike enterovírusových infekcií.

Enteroviruses belonging to the family Picornaviridae are important human pathogens. Although most cases of infection caused by these viruses are asymptomatic, a wide range of clinical syndromes is observed in manifest cases. Conventional laboratory diagnostic methods based on virus isolation and identification, or on the detection of specific antiviral antibodies, are costly and time consuming. Therefore, they are of little benefit to treatment. We have implemented a commercially available PCR-based test for the detection of enteroviral infections. In 2008, we analyzed biological specimens from 125 patients with suspected enteroviral disease, most often involving the nervous system. The presence of enterovirus was detected in 39 patients. The results were compared with those obtained by the conventional methods. PCR appeared to be a valuable method for rapid, accurate and reliable diagnosis of enteroviral infections which is of major benefit to patient management.

[Circulation of coxsackieviruses in the Slovak population in 1985-2007]. / Cirkulácia coxsackievírusov v slovenskej populáicii v období rokov 1985 az 2007.

Enteroviral infections are of the most common infections in the human population. Circulation of these viruses is high in population and they cause a wide range of clinical syndromes in human. The aim of this study was to compare the coxsackievirus circulation intensity in the population of the Slovak Republic during the period of last 23 years and to identify changes also in relation to modification in polio vaccination scheme. Marker indicating suffered infection was the presence of virus specific antibodies, assessed by means of the virus-neutralizing test. High frequency of infections by all studied coxsackievirus serotypes was confirmed in this study. Consecutive decrease in rate of anti-coxsackievirus antibodies was identified. This may be a consequence of hygiene standard improvement. After modification of immunization scheme (2004/2005) the decrease has been stopped, a mild increase of seropositivity to all studied serotypes was observed, respectively.

Coxsackievirus infection of mice. I. Viral kinetics and histopathological changes in mice experimentally infected with coxsackieviruses B3 and B4 by oral route.

We followed the viral kinetics and histopathological changes in different organs of immunocompetent mice infected orally with coxsackieviruses B3 (CVB3) Nancy strain and B4 (CVB4) JVB strain separately. The viruses used were not adapted to mouse organs. In the acute phase of infection, the viral kinetics indicated virus replication in the heart, spleen, thymus, pancreas, and small and large intestines. This was accompanied by histopathological changes, mild infiltration of mononuclear cells and fibrosis in the heart. The necrotic changes with mononuclear infiltration and fibrosis in the myocard was observed on days 56 and 71 p.i. in the CVB4-infected animals only. In the mice infected with CVB3 and CVB4 a prolonged presence of infectious virus was shown in the spleen and small intestine; in the latter viral antigen was localized in smooth muscles of the muscular wall immunohistochemically. This is the first report on prolonged replication of coxsackieviruses (CV) in the spleen and small intestine in orally infected mice.

Coxsackievirus infection of mice. II. Viral kinetics and histopathological changes in mice experimentally infected with coxsackievirus B3 by intraperitoneal route.

The study was focused on kinetics of Coxsackievirus B3 serotype (CVB3) in different organs of Swiss albino mice following intraperitoneal (i.p.) infection. The results indicated that the virus replicated in the heart, spleen, thymus, pancreas, small and large intestines in the acute stage of the infection. Infectious virus was present in the spleen till day 35 post infection (p.i.). Histopathology of the hearts showed mild foci of infiltration of mononuclear cells in the acute stage of infection and massive inflammation of exocrine pancreas on day 5 p.i. These results, when compared to those of our previous study (Bopegamage et al., 2003), suggest that the pathogenesis of the disease may be influenced by the route of virus administration into the host.

Coxsackie viral infection and orofacial cleft.

The incidence of orofacial cleft (OC) in newborns was compared with the occurrence of virus-neutralizing antibodies to coxsackie viruses in the serum of newborns and their mothers. No significant difference was found when comparing the seropositivity rates between the group of patients and the control group of healthy newborns. If the patients were divided according to the place of residence however, marked differences occurred between the regions. The lowest incidence of both--coxsackie infection and OC was determined in the region of Bratislava and the highest in the region of Zilina. The explanation of these findings recquires a more detailed analysis of genetic background, social and hygienic status, style of life and other factors, known to influence the development of OC as multi-etiological developmental disorder. (Tab. 2, Fig. 4, Ref. 12.).

Coxsackie infection and births of future diabetic children: year, seasonality and secularity.

BACKGROUND: Environmental factors including Coxsackie virus infections are considered as one of the etiopathogenetic mechanisms in childhood insulin dependent diabetes mellitus (IDDM). HYPOTHESIS: This relationship should be reflected not only in similar seasonal and secular periodicities of Coxsackie epidemics and IDDM incidence but also in the cycling of births of future diabetic children. SUBJECTS: Seven hundred and seven healthy Slovak children randomly selected in 1985-98 were investigated for the presence of actual Coxsackie infection. The months of birth in 1970-93 of 1779 Slovak children with later IDDM were registered. AntiCoxsackie positivity was determined in 1978-91 at IDDM onset in 336 children. METHODS: AntiCoxsackie positivity was assessed by IgM antibodies and virus neutralization test. Time series data were processed by cosinor regression enabling the definition of statistical significance of trends and of cycles with tested period lengths. RESULTS: Marked autumnal peaks were found for Coxsackie infections. For births of future diabetics, both autumn and spring peaks were encountered. Moreover, seven-year cycling displayed peaks in 1990 and 1997 for Coxsackie and 1976, 1983 and 1990 for IDDM births. CONCLUSIONS: Connection between Coxsackie viral infections and births of future diabetic children appears plausible also in Slovakia.

[Insulin-dependent diabetes mellitus as a possible sequelae of treatment of viral hepatitis with interferon-alpha] . / Inzulín-dependentný diabetes mellitus ako mozný následok liecby vírusových hepatitíd interferónom alpha.

Treatment of chronic hepatitis with interferon-alpha is an increasingly used successful therapeutic procedure. In the literature in recent years data accumulated on side-effects of interferon therapy, among which relatively frequently insulin dependent diabetes is mentioned. Interferon-alpha enhances the expression of molecules of the histocompatible complex I which may cause in genetically predisposed subjects the clinical manifestation of diabetes. It is therefore recommended to monitor before the onset of treatment and during interferon treatment the auto-antibody formation against islet cells and against insulin which signalizes changes in the pancreas before the clinical disease proper.

Detection of coxsackie B virus infection using a rapid screening method.

A Western blot method (WB) was adapted for rapid screening of antibodies against coxsackie virus B1-B6 in sera from patients with newly diagnosed insulin-dependent diabetes mellitus, myocarditis or febrile syndrome of suspected coxsackie viral aetiology. The use of a mixture of all 6 coxsackie virus B serotypes as the common antigen permitted a very rapid and inexpensive detection of antibody-positive sera for preliminary diagnosis and further detailed assay. Comparison of the results with those obtained in parallel run virus-neutralization tests showed a higher sensitivity and comparable specificity of WB.

Serotypic characterization of group A rotaviruses associated with children's diarrhea in Slovakia.

A total of 368 rotavirus RNA-positive (PAGE) stool samples collected continually during 1992-95 from infants and young children under five years of age hospitalized with acute gastroenteritis were serotyped using an enzyme immunoassay with VP7-specific monoclonal antibodies (ELISA with MAbs) for serotypes G1-G4. The serotype was identified in 106 stool samples (29%). Comparison of electropherotype and serotype profile in individual samples did not show any remarkable correlation. The members of three electropherotypes (A, C, K) belonged to all 4 serotypes. The representatives of two electropherotypes (E, G) and of mixed electropherotype did not react with any of the specific monoclonal antibodies used. The distribution of the serotypes was scored as 52, 13, 14 and 7.5% for G1 G4, respectively, whereas 13% of samples reacted with two or more type-specific monoclonal antibodies. The G1 serotype dominated during the period followed.

[An unusual isolation of Neisseria meningitidis]. / Neobvyklá izolácia Neisseria meningitidis.

The authors describe a unique case of isolation of Neisseria meningitidis C:2a:P1.2,P1.5 in August 1997 from a 7-year-old female patient from a vaginal smear during purulent inflammation of the external genital following injury. This is the first described case of isolation of N. meningitidis from this site described in our country. The authors draw attention to the necessity to consider uncommon occurrences of N. meningitidis, and to examine atypical materials also under the microscope and to identify by biochemical methods all isolated Neisseriae incl. gonococci.

[Parvovirus B19 antibodies in the blood in 17-34-year-old women]. / Protilátky voci parvovírusu B19 v sérach 17-34-rocných zien.

In order to assess the proportion of women in fertile age who lack antibodies against parvovirus B19 the authors assessed in 2 groups of sera (63 and 40 sera) taken from 17-34-year-old women antibodies against this virus by the ELISA method (Seiken). Virus-specific antibodies, class IgG were detected only in 67% of the examined subjects which indicates that in one third of the women there is a risk of transplacental infection of the foetus in case of infection with the parvovirus B19 during pregnancy.

Tumour necrosis factor alfa and glucose levels in sera of mice infected with coxsackie B4 and A7 viruses.

Cytokines have been suggested to play an important role in the pathogenesis of type I diabetes mellitus through their direct and indirect effects on the pancreatic islet cells. We studied the time course of tumour necrosis factor alpha (TNF-alpha) and glucose levels in the sera of mice infected with coxsackie B4 and A7 viruses. Two correlating peaks of TNF-alpha and glucose were found. These results suggest the involvement of TNF-alpha in the damage of the insulin producing cells and thus an immunity-related inflammatory process.

[Western blotting in the serologic diagnosis of enterovirus infections. I. Detection of coxsackie virus antibodies in reference sera and sera of experimentally infected mice]. / Pouzitie metódy Western blotting na sérologickú diagnostiku enterovírusových infekcií. I. Detekcia anti-coxsackievírusových protilátok v referencných diagnostických sérach a v sérach experimentálne infikovaných mysí.

The introduction of rapid serological diagnosis of enterovirus infections is faced with the problem of inter-type cross-reactions. Therefore the method of choice is still the virus-neutralization test on cell cultures and on suckling mice resp., i.e. methods which are expensive and time consuming and demanding on laboratory equipment. The detection of antibodies against the Coxsackie virus by means of Western blotting revealed a specific reaction of commercial hyperimmune diagnostic rabbit as well as of sera obtained after oral infection of mice. The application of the method in the diagnosis of human enterovirus infections will be the subject of the subsequent part of this study.

[Western blotting in the serologic diagnosis of enterovirus infections. II. Detection of coxsackievirus antibodies in the sera of patients with suspected enterovirus infection and in paired specimens of sera and saliva]. / Pouzitie metódy Western blotting na sérologickú diagnostiku enterovírusových infekcií. II. Detekcia anti-coxsackievírusových protilátok v sérach pacientov so suspektnou enterovírusovou infekciou a v párových vzorkách sér a slín.

Detection of anti-Coxsackie virus antibodies in sera with suspect enterovirus infection by the Western blotting (WB) method revealed complete agreement of results with virus neutralization tests (VNT) in those patients where in the VNT no antibodies against any of the tested Coxsackie viruses were detected. Comparison of the results of WB and VNT in other patients did not rule out an inter-type cross-reaction. In paired specimens of sera and saliva comparable results were obtained by WB, VNT and the complement fixation method. The rapid and relatively simple character of the WB method makes this method suitable for more detailed elaboration and introduction into routine diagnosis of enterovirus infections.

Characterization of human rotaviruses isolated from symptomatic and asymptomatic infections.

The occurence and circulation of different human rotavirus electropherotypes during 1984-1992 in diarrhoeic and healthy subjects were studied using examination of electropherotypes of viral RNA by polyacrylamide gel electrophoresis (PAGE). In 638 of 934 faecal specimens 11 different electropherotypes were distinguished. One long electropherotype pattern was far more frequent (58.6%) than any of the other 10 rotavirus electropherotypes detected throughout the whole study. The broadest spectrum of electropherotypes was detected in the group of children hospitalized with gastroenteritis as compared to the other groups followed. In the group of asymptomatic newborns only single electropherotype was detected, which did not occur in any group of sick children or adult contancts. Hospital-acquired infection was proved in 7.9% of followed hospitalized gastroenteritis cases. All electropherotypes detected in hospitalized children were identical to the strains occurring among community-acquired rotavirus infections. According to these findings we do not suppose that a hospital is a reservoir of rotaviruses responsible for clinically apparent gastroenteritis cases.

In vitro infection of mouse pancreatic islet cells with coxsackie viruses.

We have demonstrated the ability of 4 standard coxsackie viruses (B4, B5, A7, and A9) and one fresh isolate (A7) from a newly diabetic child with homologous serological response, to infect in vitro grown mouse pancreatic islet cells. Up to the 9th day after infection the multiplication of viruses in the cells was proved using virus titration and immunofluorescence test. Isolated pancreatic cells proved to be a suitable model for detailed studies of experimental infection of pancreatic cells with coxsackie viruses.

Isolation of group A human rotaviruses from faecal specimens in monkey MA-104 cells.

Human rotaviruses present in faecal specimens of patients with gastroenteritis were isolated and adapted to growth in MA-104 cells in roller tubes using trypsin pretreatment. Storage temperature-dependent differences were found in the extent of cytopathic effect. Multiplication of viruses in cell cultures was confirmed by immunoelectron microscopy and ELISA. Electrophoresis of rotavirus RNA in polyacrylamide gel was used to verify the identity of the electropherotypes determined in original faecal specimens and in final cell passages.

[The IMU-LA-TEST ROTA Dot-ELISA test kit for diagnosis of rotavirus infections]. / Testovanie súpravy IMU-LA-TEST ROTA Dot-ELISA na diagnostiku rotavírusových infekcií.

The newly developed IMU-LA-TEST ROTA Dot-ELISA kit manufactured in cooperation by Lachema Brno and VUVeL Brno for detection of viral proteins of the inner and outer capsid proved suitable for the rapid virological diagnosis of rotaviruses group A. The advantages of the method are that it is simple and unpretentious and does not require the use of a spectrophotometer. For comparison ROTA-ELISA Bioveta, ROTALEX Orion and PAGE were used.

[The role of coxsackie virus infection in the development of type 1 diabetes mellitus and its effect on the postinitial course of the disease in childhood]. / Spoluúcast' infekcie vírusmi Coxsackie na vývoji diabetes mellitus typ 1 a vplyv na d'al'sí postiniciálny priebeh ochorenia v detskom veku.

The authors present the results of a three-year investigation of Coxsackie viruses in children diabetics. In paired serum samples of 66 newly diagnosed patients of virus neutralizing antibodies against eight types of Coxsackie viruses were estimated. Concurrently specific antibodies of class IgM were assessed and compared with a control group of 218 healthy children. Infection with Coxsackie viruses, in particular types B2, B4 and B5, were found in 38 diabetics (in 57.6%) as compared with 6% in healthy children. The findings indicate an evidence that diabetics with Coxsackie infection at the betics. The prospective three-year investigation provided evidence that diabetics with Coxsackie infection at the time of manifestation of the disease had a worse postinitial course. Their C-peptide levels were significantly lower (p less than 0.05). The frequency and intensity of partial remission were significantly worse in the group of diabetics who had just overcome a Coxsackie virus infection (p less than 0.02).