Zástupci virové čeledě Polyomavirae jsou v lidské populaci hojně rozšířeni. Podle sérologických studií jsou téměř všichni dospělí jedinci infikováni nejméně jedním zástupcem této skupiny virů. K primární infekci dochází většinou v dětství bez jakýchkoli klinických příznaků a po primární infekci viry ustavují perzistentní infekci doprovázenou příležitostnou replikací a vylučováním viru. U imunosuprimovaných jedinců dochází k častým reaktivacím, které pak pouze u některých pacientů způsobující komplikace. Tato vlastnost lidských polyomavirů značně ztěžuje laboratorní diagnostiku. V příspěvku jsou uvedeny metody diagnostiky lidských polyomavirů, které se běžně používají pro screening, a metody, které jsou zatím ve stadiu studií, ale mají potenciál zlepšit či zpřesnit vyšetření a identifikovat pacienty v riziku vzniku onemocnění asociovaná s polyomavirovou infekcí.
The members of the viral family Polyomavirae are widespread in the human population. According to serological studies, almost all adults are infected with at least one of this group of viruses. The primary infection usually occurs in childhood without any clinical signs, and after the primary infection, the viruses establish a persistent infection accompanied by occasional reactivation and shedding of the virus. These viruses often reactivate in immunosuppressed individuals, but only in a minority of these patients, the reactivation results in disease development. This biological property of human polyomaviruses makes laboratory diagnosis considerably difficult. The paper provides an overview of methods for diagnosing human polyomaviruses, which are commonly used for screening, and methods that are still validated by research but have the potential to improve detection and to identify patients at risk of developing diseases associated with polyomavirus infection.
Active infection with BK polyomavirus (BKPyV) may cause serious complications in transplantation settings. Recently, the level of BKPyV IgG seroreactivity in graft donors has been shown to predict viremia and BKPyV-associated nephropathy in kidney transplant (KTx) recipients. Pretransplantation testing of the donor and recipient BKPyV serostatus could, therefore, identify patients at high risk. For the development of serological immunoassays, antibody response to the predominant BKPyV subtypes (BKPyV-I and BKPyV-IV) was studied using virus-like particle (VLP)-based enzyme-linked immunosorbent assay (ELISA). VLPs made from the capsid protein, VP1, derived from BKPyV-I and BKPyV-IV subtypes were produced using a baculovirus expression system and used as antigens. The tests were used for IgG antibody determination in 50 KTx recipients and 111 healthy blood donors. While 87% of samples reacted with mixed BKPyV-I and BKPyV-IV antigens, only 49% of samples were reactive in both ELISA tests when using BKPyV-I or BKPyV-IV antigens separately. Twenty-seven percent of healthy blood donors and 26% of KTx recipients were reactive only with BKPyV-I, while 9% and 20% were reactive only with BKPyV-IV, respectively. To determine the specificities of the antigens, selected seropositive samples were retested after preadsorption with soluble BKPyV-I, BKPyV-IV, or JC polyomavirus antigens. The experiments confirmed that recombinant VP1 VLP-based ELISAs predominantly detected BKPyV type-specific antibodies. The results imply that anti-BKPyV antibody ELISA tests should contain a mixture of subtype-specific VLP-based antigens instead of antigen derived from the most prevalent BKPyV-I subtype. The tests can be used for serological surveys of BKPyV infection and improved KTx patient management.
- MeSH
- ELISA metody MeSH
- imunoglobulin G krev MeSH
- lidé MeSH
- polyomavirové infekce epidemiologie MeSH
- příjemce transplantátu * MeSH
- protilátky virové krev MeSH
- séroepidemiologické studie MeSH
- transplantace ledvin * MeSH
- virus BK imunologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
Human polyomaviruses such as JC polyomavirus and BK polyomavirus have long been well known pathogens of immunocompromised patients. Several new members of this viral family have been described during the last decade. Human polyomavirus 9 seems to be a novel pathogen of transplanted patients according to some studies. The aim of our study was to determine the presence of human polyomavirus 9 in patients after kidney or stem cell transplantation (SCT) at the University Hospital in Hradec Kralove, Czech Republic. Overall 100 patients, 65 after kidney transplantation and 35 after SCT, were included into the study. At least three follow-up samples from each patient were examined for human polyomavirus 9 DNA presentation with the two previously described in-house PCR protocols. Despite the frequent reactivation of human CMV (14.3% in kidney transplantation and 63.3% after SCT) or BK polyomavirus in our patient group, there was no positivity for human polyomavirus 9 either in blood samples or urine samples. One of the possible reasons for this discrepancy versus previous published studies could be a relatively low proportion of patients treated by induction therapy before kidney transplantation in our study cohort.
- MeSH
- DNA virů genetika MeSH
- dospělí MeSH
- hostitel s imunodeficiencí * MeSH
- kohortové studie MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- nemocnice univerzitní MeSH
- polymerázová řetězová reakce MeSH
- Polyomaviridae genetika izolace a purifikace patogenita MeSH
- polyomavirové infekce epidemiologie virologie MeSH
- senioři MeSH
- transplantace kmenových buněk škodlivé účinky MeSH
- transplantace ledvin škodlivé účinky MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika epidemiologie MeSH
Human polyomaviruses HPyV6, HPyV7, TSPyV, HPyV9, MWPyV, and KIPyV have been discovered between 2007 and 2012. TSPyV causes a rare skin disease trichodysplasia spinulosa in immunocompromised patients, the role of remaining polyomaviruses in human pathology is not clear. In this study, we assessed the occurrence of serum antibodies against above polyomaviruses in healthy blood donors. Serum samples were examined by enzyme-linked immunoassay (ELISA), using virus-like particles (VLPs) based on the major VP1 capsid proteins of these viruses. Overall, serum antibodies against HPyV6, HPyV7, TSPyV, HPyV9, MWPyV, and KIPyV were found in 88.2%, 65.7%, 63.2%, 31.6%, 84.4%, and 58%, respectively, of this population. The seroprevalence generally increased with age, the highest rise we observed for HPyV9 and KIPyV specific antibodies. The levels of anti-HPyV antibodies remained stable across the age-groups, except for TSPyV and HPyV9, where we saw change with age. ELISAs based on VLP and GST-VP1 gave comparable seroprevalence for HPyV6 antibodies (88.2% vs.85.3%) but not for HPyV7 antibodies (65.7% vs. 77.2%), suggesting some degree of crossreactivity between HPyV6 and HPyV7 VP1 proteins. In conclusion, these results provide evidence that human polyomaviruses HPyV6, HPyV7, TSPyV, HPyV9, MwPyV, and KIPyV circulate widely in the Czech population and their seroprevalence is comparable to other countries.
- MeSH
- dárci krve * MeSH
- dospělí MeSH
- hostitel s imunodeficiencí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- myši MeSH
- polyomavirové infekce epidemiologie imunologie virologie MeSH
- Polyomavirus klasifikace genetika imunologie MeSH
- protilátky virové krev MeSH
- senioři MeSH
- séroepidemiologické studie MeSH
- virion imunologie izolace a purifikace MeSH
- virové plášťové proteiny genetika imunologie MeSH
- zdraví dobrovolníci pro lékařské studie MeSH
- zkřížené reakce MeSH
- zvířata MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- myši MeSH
- senioři MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Československo epidemiologie MeSH
Merkel cell polyomavirus (MCPyV) is associated with Merkel cell carcinoma (MCC), a rare skin malignancy. Human polyomavirus six and seven (HPyV6 and HPyV7) were identified on a skin but have not been associated with any pathology. The serology data suggest that infection with polyomaviruses occurs in childhood and they are widespread in population. However, the site of persistent infection has not been identified. Altogether, 103 formalin-fixed paraffin-embedded (FFPE) specimens and five fresh frozen tissues (FF) of non-malignant tonsils and 97 FFPE and 15 FF samples of tonsillar carcinomas were analyzed by qPCR for the presence of MCPyV, HPyV6, and HPyV7 DNA. All MCPyV DNA positive FF tissues were screened for the expression of early viral transcripts. Overall prevalence of MCPyV, HPyV6, and HPyV7 in non-malignant tonsillar tissues was 10.2%, 4.6%, and, 0.9%, respectively. The prevalence of MCPyV DNA in non-malignant tonsils increased with age (P < 0.05). While the prevalence of MCPyV DNA was significantly higher in the tumors than non-malignant tissues (35.7% vs. 10.2%) (P < 0.001), the prevalence of HPyV6 DNA (5.4% vs. 4.6%) and HPyV7 DNA (1.8% vs. 0.9%) were comparable. In all MCPyV DNA positive FF tissues early transcripts were detected. MCPyV, HPyV6, and HPyV7 DNAs were found in tonsils, suggesting that the tonsils may be a site of viral latency. The viral load was low indicating that only a fraction of cells are infected. The higher prevalence of MCPyV DNA was detected in tonsillar tumors but there was no difference in the viral load between tumor and healthy tissues.
- MeSH
- dospělí MeSH
- infekce onkogenními viry epidemiologie virologie MeSH
- krční mandle virologie MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- lidé středního věku MeSH
- lidé MeSH
- Merkelův nádor virologie MeSH
- polyomavirové infekce epidemiologie virologie MeSH
- Polyomavirus klasifikace izolace a purifikace MeSH
- prevalence MeSH
- RNA virová analýza MeSH
- senioři MeSH
- tonzilární nádory virologie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: Young age and thiopurine therapy are risk factors for lymphoproliferative disease among patients with inflammatory bowel disease (IBD). AIMS: The aims of this study were to evaluate the prevalence of seropositivity for the Epstein-Barr virus (EBV) and human cytomegalovirus (CMV) among children and adolescents with IBD, to assess the viral load of EBV, CMV, and BK and JC polyomaviruses (BKV, JCV) in these patients, and to assess the influence of different therapeutic regimens on seroprevalence and viral load. METHODS: Children who had been followed in our center were tested for EBV, CMV, BKV, and JCV in a cross-sectional study. One hundred and six children were included who had Crohn's disease (68%), ulcerative colitis (29%), and unclassified IBD (3%). RESULTS: We found that 64% of patients were EBV seropositive. The proportion of EBV seropositive patients increased during childhood. Azathioprine therapy (p = 0.003) was associated with EBV seropositivity in a multiple logistic regression model, after adjusting for gender, age, and disease activity at determination. We found a significant association between the number of polymerase chain reaction copies and infliximab dose (p = 0.023). We did not find any significant association between CMV serology and CMV, BKV, or JCV viral load, or any other therapeutic regimen or clinical characteristics. CONCLUSIONS: Treatment with azathioprine appears to be a risk factor for early EBV seropositivity in children with IBD, and the infliximab dose was associated with a higher EBV viral load.
- MeSH
- azathioprin škodlivé účinky MeSH
- Crohnova nemoc diagnóza farmakoterapie epidemiologie imunologie MeSH
- cytomegalovirové infekce diagnóza epidemiologie imunologie virologie MeSH
- dítě MeSH
- hostitel s imunodeficiencí MeSH
- imunosupresiva škodlivé účinky MeSH
- infekce virem Epsteina-Barrové diagnóza epidemiologie imunologie virologie MeSH
- infliximab škodlivé účinky MeSH
- lidé MeSH
- logistické modely MeSH
- mladiství MeSH
- multivariační analýza MeSH
- odds ratio MeSH
- oportunní infekce diagnóza epidemiologie imunologie virologie MeSH
- polyomavirové infekce diagnóza epidemiologie imunologie virologie MeSH
- předškolní dítě MeSH
- prevalence MeSH
- průřezové studie MeSH
- rizikové faktory MeSH
- séroepidemiologické studie MeSH
- sérologické testy MeSH
- ulcerózní kolitida diagnóza farmakoterapie epidemiologie imunologie MeSH
- věkové faktory MeSH
- virová nálož MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- mladiství MeSH
- mužské pohlaví MeSH
- předškolní dítě MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
JC and BK polyomaviruses (JCV and BKV) infect humans and can cause severe illnesses in immunocompromised patients. Merkel cell polyomavirus (MCPyV) can be found in skin carcinomas. In this study, we assessed the occurrence of serum antibodies against MCPyV, BKV, and JCV polyomaviruses in a healthy population of the Czech Republic. Serum samples from 991 healthy individuals (age range: 6-64 years) were examined by enzyme-linked immunoassay (ELISA) using virus-like particles (VLPs) based on the major VP1 capsid proteins of these viruses. Overall, serum antibodies against MCPyV, JCV, and BKV were found in 63%, 57%, and 69%, respectively, of this population. For all three viruses, these rates were associated with age; the occurrence of antibodies against MCPyV and JCV was highest for those older than 59 years, while the occurrence of antibodies against BKV was highest in those aged 10-19 years and 20-29 years. This is the first large study to determine the seroprevalence rates for BKV, JCV, and MCPyV polyomaviruses in the general Czech Republic population.
- MeSH
- buněčné linie MeSH
- dítě MeSH
- dospělí MeSH
- infekce onkogenními viry krev epidemiologie virologie MeSH
- kojenec MeSH
- lidé středního věku MeSH
- lidé MeSH
- Merkelův polyomavirus imunologie MeSH
- mladiství MeSH
- mladý dospělý MeSH
- myši inbrední ICR MeSH
- polyomavirové infekce krev epidemiologie virologie MeSH
- předškolní dítě MeSH
- protilátky virové krev MeSH
- séroepidemiologické studie MeSH
- studie případů a kontrol MeSH
- věkové rozložení MeSH
- virus BK imunologie MeSH
- virus JC imunologie MeSH
- zvířata MeSH
- Check Tag
- dítě MeSH
- dospělí MeSH
- kojenec MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- předškolní dítě MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
Polyomavirus BK (BKV) is a common human polyomavirus that rarely causes clinical symptoms in immunocompetent individuals. However, BK virus reactivation occurs in 20-40% of kidney transplant patients and 1-10% of cases present with BK virus-associated nephropathy (BKVN) and reduced kidney allograft survival. In this study, 120 consecutive renal allograft recipients were monitored for BK virus replication by real-time PCR (qPCR) in the blood and urine during the first year post-transplantation and risk factors for BK viremia, viruria, and polyoma BKV-associated nephropathy were evaluated. Receiver operating characteristic curve analysis was used to determine the cutoff points for assessing the risk of developing BKVN. In total, 1,243 samples were tested. BK-DNAuria >10(7) copies/ml and BK-DNAemia >10(4) copies/ml were found in 25.8% and 5% of the samples screened, respectively, during the 12 month follow-up period. BKVN was confirmed histologically in 3/120 patients and viremic patients were treated with dialysis for longer time periods and had higher levels of panel [corrected] reactive antibodies. Patients with viruria were also treated longer with dialysis and had impaired graft function 12 months post-transplantation. Patients with sustained viruria exhibited more acute rejection episodes than patients with transient viruria. Using receiver operating characteristic curve analysis, the cutoff point for viremia and viruria was redefined to 10(3) copies/ml serum for BK viremia and a cutoff point of 6.7 × 10(7) copies/ml in urine. In conclusion, polyoma BK viremia and viruria are frequent findings in kidney transplant recipients that warrant intensive monitoring as a means of preventing graft failure [corrected].
- MeSH
- dialýza ledvin škodlivé účinky MeSH
- dospělí MeSH
- krev virologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- longitudinální studie MeSH
- mladiství MeSH
- mladý dospělý MeSH
- moč virologie MeSH
- polyomavirové infekce diagnóza epidemiologie virologie MeSH
- prognóza MeSH
- prospektivní studie MeSH
- rizikové faktory MeSH
- ROC křivka MeSH
- senioři MeSH
- transplantace ledvin škodlivé účinky MeSH
- transplantace MeSH
- virus BK izolace a purifikace MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- hepatitida epidemiologie virologie MeSH
- HIV infekce epidemiologie přenos MeSH
- kontrola infekčních nemocí MeSH
- lidé MeSH
- Poliovirus imunologie MeSH
- polyomavirové infekce epidemiologie MeSH
- pravé neštovice epidemiologie imunologie MeSH
- ptačí chřipka u ptáků epidemiologie MeSH
- syndromy imunologické nedostatečnosti imunologie komplikace virologie MeSH
- Check Tag
- lidé MeSH
