[The role of virological tests in the diagnosis of cytomegalovirus infection in pregnant women]. / A virológiai vizsgálatok szerepe a terhes nok cytomegalovirusfertozésének kiderítésében.

INTRODUCTION: The most harmful and most frequent foetal agent is cytomegalovirus. The progress in diagnostic tools and therapeutic opportunities opened new perspectives in the diagnosis and management of foetal cytomegalovirus infection. AIM: Evaluation of cytomegalovirus virological test results performed during pregnancy between 2007 and 2012. METHOD: Clinical and virology data were retrospectively analysed. RESULTS: 64.5% of the 956 tested women were serologically protected and 33.3% were susceptible to cytomegalovirus. Recent infection was confirmed in 10 pregnant women, while the infection could not be confirmed or excluded in 3 pregnant women. Six pregnant women were asymptomatic, 5 had typical disease, and 2 had abnormal fetal ultrasound. One fetus aborted, congenital infection was confirmed in 2, and was excluded in one of the four newborns tested. CONCLUSIONS: The immunity of women to cytomegalovirus reflects high socioeconomic circumstances. Confimatory tests must be done both in women who have cytomegalovirus disease and those who have IgM positive result detected by enzyme (linked) immunoassay. Screening must be done prior to pregnancy. Strict collaboration between professionals of different medical specialties is necessary.

[Observations on human parvovirus B19 infection diagnosed in 2011]. / A 2011. évi B19 humán parvovírus-fertozésekrol szerzett tapasztalatok.

INTRODUCTION: The incidence of human parvovirus B19 infection is unknown. AIM: A retrospective analysis of clinical and laboratory findings was carried out in patients diagnosed with human parvovirus B19 infection in 2011 in a virologic laboratory of a single centre in Hungary. METHODS: Clinical and laboratory data of patients with proven human parvovirus B19 infection were analysed using in- and out-patient files. RESULTS: In 2011, 72 patients proved to have human parvovirus B19 infection with the use of enzyme immunoassay. The clinical diagnoses of these patients were as follows: human parvovirus B19 infection (30.6%), transient aplastic crisis (16.7%), arthritis (8.3%) and acute hepatitis (4.1%). Symptoms of each of the four phases of the infection occurred in various combinations with the exception of the monophase of cheek exanthema. This occurred without the presence of other symptoms in some cases. Leading symptoms and signs were exanthema (in 74.6% of cases), haematological disorders (in 69% of cases), fever (in 54.9% of cases) and arthritis (in 33.8% of cases). Several atypical dermatological symptoms were also observed. Acute arthritis without exanthema was noted in 8 patients. Of the 72 patients with proven human parvovirus B19 infection there were 7 pregnant women, and one of them had hydrops foetalis resulting spontaneous abortion. In 16 patients (22.5%) human parvovirus B19 IgG was undetectable despite an optimal time for testing. CONCLUSION: The observations of this study may contribute to a better recognition of clinical symptoms of human parvovirus B19 infection.

[Clinical significance and the first identification of human parechoviruses in Hungary]. / A humán parechovírusok klinikai jelentosége és elso hazai azonosítása.

UNLABELLED: Human parechoviruses (HPeV) belonging to the family Picornaviridae are widespread enteric pathogens and are associated with various clinical syndromes in human. At present, 16 HPeV genotypes (HPeV1-16) are known. There is no report on the detection of HPeVs in Central Europe. AIMS: The aim of the retrospective study was to detect and characterize HPeVs using molecular methods in cell cultures with "enterovirus-like" cytophatic effect (CPE) archived between 1990 and 2004, in two virology laboratories, in Hungary. MATERIALS AND METHODS: In Laboratory I, fecal samples from children with symptoms of gastroenteritis under the age of 10 years were cultured as a previous routine diagnostic laboratory protocol for "enterovirus". Cell cultures indicating CPE were archived between 1990 and 2000. In Laboratory II, 2 fecal samples, a liquor and a nasopharyngeal aspirate were re-tested which contained an "enterovirus-like" virus in cell cultures and were positive by HPeV1 neutralization immunosera between 2000 and 2004. Specimens were tested retrospectively for HPeV by reverse transcription-PCR (RT-PCR) method using 5'UTR conserved primers. Specific primers were designed to determine the HPeV structural region (VP0-VP3-VP1). RESULTS: 9 of the 66 archived samples (9.1%) from Laboratory I and all the 4 samples from Laboratory II were found to be HPeV-positive. 10 samples were identified as HPeV1, 2 were HPeV4 and 1 could not be determined. 3 HPeV1 clusters were identified in Laboratory I according to the isolation date originated from years 1990/1991, 1992/1995 and 1998. HPeV1 was detected in clinical syndromes: gastroenteritis (in a 24-years-old adult), recurrent stomatitis aphtosa (in a 42-years-old adult), encephalitis and ataxia cerebellaris acuta in infants and children in Laboratory II. CONCLUSIONS: This is the first detection of HPeVs in Central Europe. Detection and genetic characterization of HPeV in available historical samples infected with previously unidentifiable agents with "enterovirus-like" cytopathogenic effect may help to understand the clinical importance and spectrum of the infections and the genetic diversity and evolution of these viruses.

Trends in the epidemiology of human G1P[8] rotaviruses: a hungarian study.

Epidemiological trends of the globally most common rotavirus genotype, G1P[8], were investigated in Hungary during a 16-year period by sequencing and phylogenetic analysis of the surface antigens. Antigen shift among epidemiologically major G1P[8] strains was observed in 6 seasons, as indicated by changes in the sublineages of the G1 VP7 and the P[8] VP4 genes. The temporal clustering of some rotavirus VP4 and VP7 gene sublineages and the periodic emergence and/or resurgence of previously unrecognized rotavirus sublineages in the study population suggest a dynamic nature for these common strains. Recently established international strain surveillance networks may help to identify and track the spread of epidemiologically important rotavirus strains across countries and continents.

[Experience with multiplex nested PCR and fluorescent antibody tests in the diagnosis of acute central nervous system infections with herpes simplex virus type 1 and 2]. / Tapasztalatok a heveny központi idegrendszeri fertozések herpes simplex vírus-1/2 diagnosztikájában a multiplex nested PCR- és a fluoreszcens jelzésu antitestválasz-vizsgálat kombinációjával.

UNLABELLED: The specific diagnosis of herpes simplex virus type 1 and 2 infections has an extreme importance in acute infections of central nervous system due to both availability of specific antiviral therapy and the possible serious consequences of the disease. AIMS: Evaluation of the relevance and interpretation of the results of PCR and the specific antibody testing. METHODS: Home made multiplex nested herpes simplex virus PCR and immunofluorescent IgM, IgA, IgG antibody tests were carried out in a total of 474 cerebrospinal fluid and 555 serum samples of 396 patients with acute infection of the central nervous system between 1. January, 2003 and 31. December, 2009. RESULTS: The herpes simplex virus etiology was verified in 21% of 396 patients (82 patients, mean 12 cases per year): 26 were diagnosed by both methods (32%), 41 by PCR only (50%), 15 by the detection of intrathecal antibody production only (18%) (p<0.0001). HSV type1 or 2 DNA remained detectable in 35% of the samples drawn after the 30th day of the disease. These patients were all younger than two years of age. CONCLUSIONS: 1. PCR increased the ratio of verified herpes simplex virus etiology in acute central nervous infections. 2. Testing the specific antibody response cannot be ceased even in the availability of PCR. 3. Herpes simplex virus type 1 or 2 DNA might persist in central nervous system in spite of the specific antiviral therapy especially in the infants. 4. Herpes simplex virus PCR can be repeated if an early sample is negative or if it is suspected false positive. 5. There is a need for cooperation between clinicians and virologists in the appropriate interpretation of the results and in finding etiology.

[Post vaccination rotavirus surveillance in Hungary, in 2007]. / Posztvakcinációs rotavírus-surveillance Magyarországon, 2007.

Vaccination is the main strategy to control severe dehydrating gastroenteritis caused by rotaviruses in early childhood. The availability of new generation rotavirus vaccines has led to an intensification of strain surveillance worldwide, in part, to gauge the impact of the possible vaccine-driven immune selection of wild-type rotavirus strains. In the present study, authors describe the strain prevalence data obtained in 2007, with the involvement of different regions of Hungary. Genomic RNA was extracted from rotavirus-positive stool samples collected mainly from children and then subjected to genotyping using multiplex RT-PCR assay. Type-specific primers targeted G1 to G4, G6, G8 to G10, and G12 VP7 specificities, and P[4], P[6], and P[8] to P[11] VP4 specificities were used. Out of 489 rotavirus-positive specimens, collected from 482 patients, 466 and 474 were successfully G and P typed, respectively, and both G and P type specificities could be assigned for 457 strains. Prevalence data showed the predominance of G4P[8] (31.5%) strains, followed by G1P[8] (28.3%), G2P[4] (19.3%), and G9P[8] (10.2%). Minority strains were G1P[4] (0.4%), G2P[8] (1.3%), G3P[9] (0.2%), G4P[6] (0.7%), G6P[9] (0.4%), G8P[8] (0.2%), G9P[4] (0.2%), G9P[6] (0.8%), and G12P[8] (0.4%). Mixed infections were found in 1.2% of the samples, while 4.9% remained partially or fully non-typified. Our data indicate that the antigen specificities of medically important rotavirus strains identified in this 1-year study are well represented in the vaccines available in the pharmaceutical private market in Hungary. Depending on the vaccination coverage achievable in the forthcoming years, the post-vaccination rotavirus strain surveillance may allow us to gain comprehensive information on the impact of rotavirus vaccines on the prevalence of circulating rotavirus strains.

[Acute disseminated encephalomyelitis in childhood]. / Akut disszeminált encephalomyelitis gyermekkorban.

BACKGROUND AND PURPOSE: Acute disseminated encephalomyelitis is a rare inflammatory demyelinating disorder often preceded by infection or vaccination. The purpose of the study was the systematic analysis of clinical, radiological and microbiological profiles of children treated at Szent László Hospital, and the comparison of findings with literature data. METHODS: Demographic, infectological, clinical, radiological, laboratory and virological data of patients treated and followed-up between 1-Jan-1998 and 30-June-2008 were reviewed and analysed. RESULTS: 19 children met diagnostic criteria. Their mean age was 6.8 years. A prodromal illness--mostly febrile viral infection, upper respiratory infection or chickenpox--preceded neurological symptoms in 17 patients. All had polysymptomatic encephalopathy, 2 children had spinal symptoms. The cerebrospinal fluid was abnormal in all but one. A viral etiology was definite in 7 and probable in 8 cases. MRI disclosed white matter changes in 18, cortical and deep gray matter in 16, cerebellar in 6, brain stem in 14 and spinal cord changes in 2 cases. Repeat MRI performed mean 4 months later showed complete resolution in 6 and partial resolution in 11 patients. 13 patients received high-dose methylprednisolone, 2 of whom were also treated with plasma exchange and 1 with immunoglobulin. 9 children required mechanical ventilation. 2 patients died, 10 recovered without and 7 with sequelae. 2 patients developed further demyelinating events: multiple sclerosis and multiphasic disseminated encephalomyelitis, respectively. CONCLUSION: Clinical, radiological and follow-up results were similar to those published in literature however, triggering viruses were identified in a larger proportion of cases.

[EuroRotaNet--European rotavirus strain surveillance network established with Hungarian participation]. / Hazai részvétellel megalakult az Európai Rotavírus-törzs-figyelô Hálózat (EuroRotaNet).

Group A rotaviruses are the most common cause of severe gastroenteritis worldwide. The incidence and distribution of group A rotavirus sero/genotypes varies between geographical areas during a rotavirus season, and from one season to the next. In addition, cocirculation of genetically diverse multitypic rotaviruses and of intratypic variants in any one place and time is common. Assuming widespread use of rotavirus vaccine in the near future, comprehensive surveillance of natural rotavirus infections is vital. EuroRotaNet has been established in order to gather comprehensive information on the rotavirus types co-circulating throughout Europe. The main objectives of the network are to (i) develop methods and algorithms for effective rotavirus strain typing and characterisation, (ii) describe in detail the molecular epidemiology of rotavirus infections in Europe, (iii) monitor the effectiveness of current genotyping methods and respond to changes associated with genetic drift and shift, and (iv) monitor the emergence and spread of novel rotavirus strains within Europe. This infrastructure may serve as a platform for future surveillance activities and nested studies for evaluating the effectiveness of a rotavirus vaccine in the general population. Studies to monitor the reduction in disease associated with common rotavirus types, the possible vaccine-induced emergence of antibody escape mutants of genotypes other than those included in the vaccine and of reassortment between vaccine and naturally circulating wildtype strains are required.

Whole genome sequencing and phylogenetic analysis of a zoonotic human G8P[14] rotavirus strain.

The full-length genome of a rare human G8P[14] rotavirus strain, BP1062/04, identified during a surveillance study in Hungary was determined and analyzed. This strain showed a G8-P[14]-I2-R2-C2-M2-A11-N2-T6-E2-H3 genomic constellation. Phylogenetic analysis of each genome segment revealed common origins with selected animal and zoonotic human strains. The closest relatedness was seen with suspect zoonotic Hungarian G6P[14] strains in the NSP1 and NSP3 gene phylogeny, with ovine strains in the VP1, VP2, NSP4 gene phylogeny, and with bovine strains in the NSP5 gene phylogeny. The outer capsid VP7 and VP4 genes could not be derived from cognate genes of any known human or animal G8P[14] strains. The remaining genes, NSP2, VP3 and VP6, gave no definite clues to the host origin, although each was clearly different from true human strains. Altogether, our findings suggest that strain BP1062/04 represents an example of a direct zoonotic transmission event.

Emergence of serotype G12 rotaviruses, Hungary.

We describe the emergence of serotype G12 rotaviruses (67 [6.9%] of 971 specimens tested) among children hospitalized with rotavirus gastroenteritis in Hungary during 2005. These findings are consistent with recent reports of the possible global spread and increasing epidemiologic importance of these strains, which may have implications for current rotavirus vaccination strategies.

Dominating prevalence of P[8],G1 and P[8],G9 rotavirus strains among children admitted to hospital between 2000 and 2003 in Budapest, Hungary.

Group A rotaviruses are the main cause of acute dehydrating diarrhea in children, responsible for high mortality in developing countries and a significant socio-economic burden associated with treating the disease in developed countries. Two rotavirus vaccine candidates predicated on either homotypic or heterotypic protection have undergone clinical trials recently and await licensure for routine use. In anticipation of a future vaccination campaign in Hungary, the diversity of rotaviruses collected from Budapest between 2000 and 2003 were analyzed by polyacrylamide gel electrophoresis (PAGE) of the viral genome and by serotyping and genotyping of the outer capsid genes, VP7 and VP4. Among 2,763 rotavirus positive specimens available for analysis, we were able to determine the electropherotype of 2,227, and, of these, 1,517 (68.1%) were subjected to G typing and 1,173 (52.7%) were subjected to P typing. We successfully G typed 1,481 (97.6%) and P typed 1,130 (96.3%) strains, respectively. A total of six G types (G1, 50.2%; G2, 2.2%; G3, 1.7%; G4, 5.8%; G6, 0.6%; and G9, 34.4%) and four P types (P[4], 3.0%; P[6], 0.7%; P[8], 89.9%; and P[9], 1.7%) were identified in nine individual combinations (P[8],G1; P[4],G2; P[8],G3; P[8],G4; P[8],G9; P[6],G4; P[4],G1; P[9],G3; and P[9],G6). The prevalence of VP7 and VP4 specificities varied from year to year. In this regard, a shift in serotype predominance from G1 in 2000-2001 (61.8%) and 2001-2002 (69.7%) to G9 in 2002-2003 (51.3%) was an intriguing observation that has been reported recently in some other countries, as well. The emergence of serotype G9 rotaviruses in Hungary and other parts of the world may have implications for future vaccine development and use, particularly, if current vaccine candidates cannot confer adequate homotypic or heterotypic protection against these strains.

Eight-year survey of human rotavirus strains demonstrates circulation of unusual G and P types in Hungary.

Between 1992 and 2000, a total of 4173 rotavirus-positive samples were collected from two areas of Hungary. Of these, 2020 specimens (48.4%) were analyzed for G serotype, using monoclonal antibody-based immunoassay and reverse transcription-PCR. By the two methods, 1789 samples were specified as G1 (62%), G2 (12.2%), G3 (1.4%), G4 (6.4%), G6 (1.0%), G9 (2.9%), or mixed infection (2.6%), and the remaining 231 (11.4%) could not be G typed. The linkage between G and P type, subgroup specificity, and RNA profile was investigated with a sample subset. Among these specimens, we identified both the four globally common strains (P[8],G1 subgroup II (sgII); P[4],G2 sgI; P[8],G3 sgII; and P[8],G4 sgII) and six uncommon strains (P[6],G4 sgII; P[9],G3 sgI; P[9],G6 sgI; P[14],G6 sgI; P[8],G9 sgII; and P[8],G9 sgI). All strains with P[8], P[6], P[9], and P[14] specificities had a long electropherotype, whereas most of those carrying a P[4] specificity were associated with a short electropherotype. Although once considered to be rare, P[9],G6 and P[8],G9 rotavirus strains represent potentially important new serotypes in Hungary.