Numerous serotypes which belong to the genus Enterovirus (EV) show variability in their virulence and clinical manifestations. They are also known to undergo changes caused by mutations and recombination during their circulation in the environment and the population. Various EV serotypes are prevalent in groundwater, wastewater and surface waters. Our previous studies showed that oral infection induces pancreatitis depending on specific conditions, such as gravidity, in an outbred murine model. Our aim in the present study was to further explore the pancreatic histopathology in an outbred mouse model following oral infection with clinical isolates from a patient who had aseptic meningitis and an isolate from a treated-sewage sample recovered from the residential area of the patient. The isolates were identified as coxsackievirus B4 (CVB4) in tissue culture. The CVB4 sewage-isolate induced pancreatitis after oral infection. In contrast, pancreatitis was absent following infection with the clinical isolates. Comparison of polyprotein sequences showed that the treated-sewage strains differed from the patient's isolates by 9 and 11 amino acids. We conclude that the isolates of clinical and environmental origin differed in their pathogenic properties and showed genetic variation.
- MeSH
- coxsackie virózy * virologie MeSH
- enterovirus B lidský * patogenita fyziologie MeSH
- lidé MeSH
- myši MeSH
- odpadní vody * virologie MeSH
- pankreatitida * chemicky indukované virologie MeSH
- virulence MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Enteroviruses (EVs) infections occur worldwide. Although, infections by these viruses are often asymptomatic and go unnoticed, they can be shed in stool for several weeks. The EVs re associated with sporadic outbreaks and a wide range of clinical symptoms, occasionally accompanied with fatal consequences. Presently in the Slovak Republic (SR) recreational waters are tested only for bacterial indicators. Our aim was to monitor EVs in recreational waters. Water samples were collected during the years 2012-2014 from different recreational natural lakes in Central and West regions of SR. The samples were concentrated by centrifugation using the two-phase separation method recommended by the World Health Organization (WHO) used for EVs surveillance in the treated sewage waste water. Each of the two phases collected from the samples was analysed by polymerase chain reaction for detection of EVs and primary sequencing was done. Our study demonstrated presence of EVs in three localities consecutively for three years, indicating a probability of constant local source of faecal contamination. This is the first monitoring report on the occurrence of EVs in the natural recreational waters in SR.
- MeSH
- Enterovirus izolace a purifikace MeSH
- feces virologie MeSH
- lidé MeSH
- mikrobiologie vody * MeSH
- monitorování životního prostředí * MeSH
- polymerázová řetězová reakce MeSH
- rekreace * MeSH
- sladká voda virologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- dopisy MeSH
- Geografické názvy
- Slovenská republika MeSH
Influenza is one of the most common human infectious diseases, and has profound health and economic consequences. The laboratory diag- nosis of influenza virus infections plays an important role in the global surveillance of influenza. Therefore, there is a growing demand for highly sensitive and rapid methods for detecting influenza. The performance of particular diagnostic methods is affected by various factors. In this study, we assess the effects of patients' age and time to diagnosis on the probability of detecting influenza using four diagnostic methods (virus isolation, rapid test, RT-PCR and real-time RT-PCR). We examined 3,546 samples from central and eastern Slovakia during the influenza seasons from 2005-2006 to 2010-2011. In general, the probability of influenza detection significantly decreased with the time from onset of illness to sample collection (T1) as well as with patients' age (AGE). On the contrary, time from sample collection to delivery (T2) did not play a role in the prob- ability of influenza detection. As judged by odds ratios, the virus isolation method was most sensitive to T1, followed by the rapid test and RT-PCR methods. For the effect of AGE, the rapid test and virus isolation methods were more sensitive than PCR-based methods. The effects of T1 and AGE were independent of each other. Laboratories which participate in inifluenza surveillance should use several methods to enable rapid and accurate influenza A and B virus detection.
- MeSH
- časové faktory MeSH
- chřipka lidská diagnóza epidemiologie MeSH
- klinické laboratorní techniky metody MeSH
- lidé MeSH
- ochrana veřejného zdraví metody MeSH
- roční období MeSH
- senzitivita a specificita MeSH
- věkové faktory MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Slovenská republika MeSH
Práca predkladá prehľad diagnostických metód vo virológii a ich využitie na Slovensku. Má poskytnúť lepšiu orientovanosť medicínskych pracovníkov v jednotlivých metodikách, ich princípoch a využití pri diagnostike vírusových infekcií. Rozvoj molekulárnych technológií značne napomohol vývoj nových molekulárno-biologických metód, ktoré zohrávajú v diagnostike, popri klasických metódach, nezastupiteľné miesto. Spoznanie princípov jednotlivých metodík pomôže klinickým pracovníkom rozšíriť ich doterajšie poznatky virologickej diagnostiky, lepšie ozrejmiť výpovednú hodnotu výsledku virologického vyšetrenia a jeho vplyvu na liečbu pacienta.
The work presents an overview of methods applied in virological diagnostics and their implementation in Slovakia. It provides general orientation in different methodologies, principles and their application in the diagnosis of viral infections to the medical personnel. The development of molecular technologies greatly progressed the use of these methods in diagnostics, which play an important irreplaceable role parallel to conventional methods. Knowledge of the principles of these methodologies will help clinicians expand their existing knowledge of the virological diagnosis, to clarify the predictive value of virological test results and their impact on patient treatment.
Odbor lekárskej mikrobiológie na Regionálnom úrade verejného zdravotníctva v Banskej Bystrici pracuje ako spádové virologické laboratórium pre stredoslovenský a v chrípkovej sezóne 2009/2010 aj pre východoslovenský región. Materiál a metódy: Vzorky (nazofaryngálne výtery a pitevné materiály) boli odoberané od sentinelových aj nesentinelových lekárov, od pacientov so suspektným chrípkovým ochorením. Vzorky boli vyšetrované rýchlotestom a následne RT-PCR metódou na diagnostiku chrípky A, resp. B. Na subtypizáciu pandemickej chrípky A/H1N1 bola použitá real-time PCR metóda. Výsledky: Od mája 2009 do júna 2010 laboratórium vyšetrilo 2497 vzoriek na prítomnosť vírusov chrípky A a B a predovšetkým na prítomnosť pandemickej chrípky A/H1N1. Z 589 vzoriek pozitívnych na chrípku A bolo 537 vzoriek subtypizovaných ako pandemická chrípka A/H1N1. Predstavuje to 21,5 % zo všetkých vyšetrených vzoriek a 91,2 % zo vzoriek pozitívnych na chrípku A. Záver: V chrípkovej sezóne 2009/2010 na strednom a východnom Slovensku jednoznačne dominovala nová pandemická chrípka A/H1N1. PCR metódy sa stali kľúčovými pri vyšetrovaní pacientov s podozrením na pandemickú chrípku v laboratóriu zapojenom do surveillance chrípky a chrípke podobných ochorení v SR.
The Department of Medical Microbiology of the Regional Authority of Public Health (RAPH) in Banská Bystrica serves as a catchment laboratory of virology for the Central Slovakia Region, and in the influenza season 2009/10, it also served as such for the East Slovakia Region. Material and methods: Specimens (nasopharyngeal swabs and post-mortem specimens) from patients with suspected influenza were obtained from both sentinel and non-sentinel physicians. The specimens were analyzed by a rapid test, followed by real-time PCR (RT-PCR) for influenza A or B diagnosis. RT-PCR subtyping for pandemic influenza A/H1N1 was performed. Results: From May 2009 to June 2010, 2497 specimens were analyzed for the presence of influenza A and B viruses and in particular for the presence of pandemic influenza A/H1N1 virus. As many as 537 of 589 influenza A-positive specimens, i.e. 21.5% of all specimens analyzed and 91.2% of influenza A-positive specimens, were subtyped as pandemic influenza A/H1N1. Conclusion: In the influenza season 2009/10, the new pandemic influenza A/H1N1 clearly predominated in Central and Eastern Slovakia. PCR tests have played a key role in diagnosing patients with suspected pandemic influenza in the laboratory participating in the surveillance of influenza and influenza-like illness in the Slovak Republic.
Many severe diseases of the respiratory tract lead to hospitalisation. These diseases are often caused by viral infections and may cause increased mortality. The most common viral pathogens involved in these cases, which are also associated with significant morbidity and mortality during the influenza seasons are influenza viruses. Rapid differential diagnosis of influenza viruses is therefore of great importance. Classical diagnosis of these viruses involves virus cultures. Of the rapid diagnostic methodologies which have been developed are RT-PCR, multiplex PCR, real-time PCR. In the present study we have monitored clinical samples from patients of different age groups from selected regions in Slovakia and compared the effectiveness of the classical and molecular biological diagnostic methods. The molecular biological methods proved to be rapid, accurate and effective. Application of these techniques in diagnosis of the respiratory illnesses should help in the prevention, therapy and disease control.
