PURPOSE: About 15% of patients with common variable immunodeficiency (CVID) develop a small intestinal enteropathy, which resembles celiac disease with regard to histopathology but evolves from a distinct, poorly defined pathogenesis that has been linked in some cases to chronic norovirus (NV) infection. Interferon-driven inflammation is a prominent feature of CVID enteropathy, but it remains unknown how NV infection may contribute. METHODS: Duodenal biopsies of CVID patients, stratified according to the presence of villous atrophy (VA), IgA plasma cells (PCs), and chronic NV infection, were investigated by flow cytometry, multi-epitope-ligand cartography, bulk RNA-sequencing, and RT-qPCR of genes of interest. RESULTS: VA development was connected to the lack of intestinal (IgA+) PC, a T helper 1/T helper 17 cell imbalance, and increased recruitment of granzyme+CD8+ T cells and pro-inflammatory macrophages to the affected site. A mixed interferon type I/III and II signature occurred already in the absence of histopathological changes and increased with the severity of the disease and in the absence of (IgA+) PCs. Chronic NV infection exacerbated this signature when compared to stage-matched NV-negative samples. CONCLUSIONS: Our study suggests that increased IFN signaling and T-cell cytotoxicity are present already in mild and are aggravated in severe stages (VA) of CVID enteropathy. NV infection preempts local high IFN-driven inflammation, usually only seen in VA, at milder disease stages. Thus, revealing the impact of different drivers of the pathological mixed IFN type I/III and II signature may allow for more targeted treatment strategies in CVID enteropathy and supports the goal of viral elimination.
- MeSH
- atrofie komplikace patologie MeSH
- běžná variabilní imunodeficience * komplikace imunologie MeSH
- CD8-pozitivní T-lymfocyty MeSH
- imunoglobulin A MeSH
- infekce viry z čeledi Caliciviridae * imunologie MeSH
- interferony MeSH
- lidé MeSH
- Norovirus * fyziologie MeSH
- zánět komplikace MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Aim: Overactivation of the IL-33/IL-13 axis is the main step in initializing allergic inflammation and promoting allergic diseases. Data on viral pathogens as risk factors for subsequent allergic disease are contradictory. The strongest associations have been made between upper respiratory tract virus infections and asthma. Intestinal viral infections also activate IL-33 and IL-13 as part of the innate antiviral response. The aim of this study was to test whether there are differences in IL-13 and IL-33 concentrations in pediatric patients with acute rotavirus- and norovirus infections and healthy controls. Material and Methods: Forty children with acute rotavirus, 27 with acute norovirus intestinal infections and 17 control children were enrolled in this study. Blood IL-33 and IL-13 detection was performed with enzyme-linked immunosorbent assays (ELISAs). Results: Acute rotavirus infection caused a significant elevation in IL-33 and IL-13 compared to acute norovirus infection (63.85 pg/ml vs. 0, P = 0.0026, and 94.24 pg/ml vs. 0.88 pg/ml, P = 0.0003, respectively) and healthy controls (63.85 pg/ml vs. 9.89 pg/ ml, P = 0.0018, and 94.24 pg/ml vs. 0.14 pg/ml, P < 0.0001, respectively). There was no significant difference in IL-33 and IL-13 concentrations between the acute norovirus group and healthy controls (0 vs. 9.89 pg/ml, P = 0.8276 and 0.88 pg/ml vs. 0.14 pg/ml, P = 0.1652, respectively). Conclusion: Acute rotavirus infection causes a significant elevation in IL-33 and IL-13, compared to norovirus and healthy control children.
Cíl práce: Noroviry patří do čeledi Caliciviridae. V současnosti jsou tyto viry klasifikovány minimálně do 10 genoskupin. Význam norovirů v České republice není zatím intenzivně sledován. Cílem studie bylo přispět ke znalostem o jejich cirkulaci v naší populaci na základě určení jednotlivých genotypů u pacientů, u kterých byla norovirová infekce diagnostikována rutinním vyšetřením během hospitalizace ve Fakultní nemocnici v Plzni. Materiál a metody: Od června 2017 do března 2020 byla diagnóza norovirové infekce potvrzena u 118 pacientů. Laboratorní vyšetření: Vzorky nativní stolice nemocných s projevy gastroenteritidy byly rutinně zpracovány kvalitativním imunochromatografickým testem RIDAQuick Norovirus (R-Biopharm AG) na stanovení noroviru genoskupiny I a II podle pokynu výrobce. Následně byla provedena molekulárně-biologická analýza vzorků. Ze vzorků stolice byla připravena 10% suspenze v roztoku fosfátového pufru, ze které byla soupravou QIAamp Viral RNA Mini Kit (Qiagen) izolována nukleová kyselina podle návodu výrobce. Dále byla provedena reverzní transkripce a amplifikace (PCR) specifické oblasti genomu norovirů (tzv. region C). PCR produkty o specifické velikosti byly přečištěny pomocí QIAquick PCR Purification Kit (Qiagen) a odeslány na sekvenování (Eurofins Genomics). Získané sekvence byly analyzovány v programu MEGA verze X. Zařazení prokázaných virů do jednotlivých genotypů bylo ověřeno s použitím Norovirus Typing Tool Version 2.0 (https://www.rivm.nl/mpf/typingtool/norovirus/). Výsledky: Během sledovaného období bylo prokázáno 14 různých genotypů, resp. variant z genoskupin GI, GII a GIX (dříve GII.15). Nejčastěji byly potvrzeny infekce vyvolané noroviry (NoV) GII.4 Sydney-2012, které vyvolaly 70,3 % onemocnění. Tato varianta norovirů byla prokázána ve 24 měsících z celkového počtu 34 měsíců sledování. Norovirové infekce se vyskytovaly především v podzimních a zimních měsících (říjen-březen), v tomto období bylo diagnostikováno 68, tj. 57,6 % onemocnění. Norovirová infekce byla potvrzena ve všech věkových skupinách, věk pacientů se pohyboval od 0 do 96 let (medián 8, průměr 27,9 let). Analýza zastoupení nejčastější varianty NoV GII.4 Sydney-2012 potvrdila statisticky významný rozdíl ve výskytu této varianty ve věkové skupině 0-4 let při porovnání se staršími osobami (χ2 = 5,37, P = 0,02). Pacienti s norovirovou infekcí měli bydliště vesměs v okrese Plzeň-město (51 osob). Z anamnézy nemocných vyplynulo, že 35 z nich (29,7 %) mělo v době prvních příznaků dalšího nemocného s příznaky gastroenteritidy v rodině. Závěr: Studie je prvním pokusem o zmapování molekulární epidemiologie norovirů nejen v Plzeňském kraji, ale v celé České republice. I přes relativně nízké počty oficiálně hlášených onemocnění patří nepochybně noroviry i u nás k velmi důležitým původcem gastroenteritid, je proto žádoucí rozšiřovat spektrum našich znalostí o jejich cirkulaci.
Objective: Noroviruses are members of the Caliciviridae family and are currently segregated into at least 10 genogroups. The distribution of these viruses in the Czech Republic has not yet been investigated in detail. A pilot study was performed to contribute to the overall knowledge and understanding of norovirus circulation in the population. Clinical specimens from patients diagnosed with norovirus infection during their hospitalization at the Pilsen University Hospital were genotyped. Material and methods: A total of 118 patients were diagnosed with norovirus infection between July 2017 and March 2020. Stool samples from patients presenting with gastroenteritis were routinely screened by the RIDA®QUICK Norovirus Test (R-Biopharm AG), a rapid chromatographic immunoassay for the qualitative detection of Norovirus genogroups I and II, according to the manufacturer’s instructions. Norovirus positive samples were subsequently analysed by molecular biological methods. Stool suspensions (10%) were prepared with phosphate-buffered saline, and nucleic acid was extracted using the QIAamp Viral RNA kit (Qiagen) according to the manufacturer’s instructions. To investigate the genotype distribution, RT-PCR targeting specific sections of the norovirus genome (region C) was employed, followed by purification of PCR products using the QIAquick PCR Purification Kit (Qiagen) and sequencing (Eurofins Genomics). The sequences obtained were analysed by the MEGA X software, and the results of phylogenetic analyses were confirmed by Norovirus Typing Tool Version 2.0 (https://www.rivm.nl/mpf/typingtool/ norovirus/. Results: During the study period, 14 norovirus genotypes or genogroup variants NoV GI, NoV GII and NoV GIX (previously NoV GII.15) were identified. The major genotype NoV GII.4 Sydney-2012 accounted for a total of 70.3% of norovirus gastroenteritis cases. This norovirus variant was detected in 24 months out of 34 months of the study period. In general, the number of norovirus infections increased during autumn and winter months (October to March) when 68 (57.6%) cases were diagnosed. Although norovirus infection was confirmed in all age categories (age range 0–96 years, median 8, mean 27.9), the statistical analysis revealed a significant difference in the incidence of NoV GII.4 infection between the age group 0–4 years and older patients (χ2 = 3.95, P = 0.047). Many patients (51) were residents of the Pilsen-city district. The case history data showed that 35 of them (29.7%) had another family member who developed symptoms of gastroenteritis at the time of the onset of their infection. Conclusion: The pilot study is the first attempt to map the molecular epidemiology of noroviruses, not only in the Pilsen Region but also in the whole Czech Republic. Despite the relatively low number of officially reported cases, noroviruses are undoubtedly one of the most important causes of gastroenteritis in this country. Further studies are therefore necessary to expand the body of knowledge of their ecology and circulation.
- MeSH
- dítě MeSH
- dospělí MeSH
- epidemiologické modely MeSH
- feces mikrobiologie MeSH
- gastroenteritida etiologie mikrobiologie MeSH
- genotyp MeSH
- infekce viry z čeledi Caliciviridae * epidemiologie MeSH
- lidé MeSH
- Norovirus izolace a purifikace MeSH
- pacienti hospitalizovaní MeSH
- Check Tag
- dítě MeSH
- dospělí MeSH
- lidé MeSH
Field-grown strawberries, the environment of strawberry farms and fresh strawberries from marketplaces were examined for bacterial, viral, and protozoan pathogens. The presence of bacteria was determined using culture and real-time PCR (qPCR), presence of protozoa and viruses using qPCR and reverse transcription qPCR, respectively. The highest proportion of positivity was observed for Escherichia coli both in field and purchased strawberries (up to 48.6%). Finding of Cronobacter ranged from 0.6% to 9% both for field and market strawberries. The prevalence of other pathogens (Listeria monocytogenes, Giardia intestinalis, Cryptosporidium sp., and Norovirus) in strawberries was below 4.5%; HAV was not detected at all. Positivity of the environment was determined to be lower than 2.1% for all microorganisms, except for E. coli. The concentration of pathogens in most samples did not exceed 100 CFU/g using culture and 1.8 × 102 GE/g of strawberries or swabbing area 6.1 × 102 GE/mL or swabbing area of environmental samples using qPCR. All studied farms applied preventive measures such as drip irrigation, avoidance of organic fertilizers, and use of mulch foils or gloves for workers to decrease contamination of strawberries. Despite this, certain pathogens were found in fresh strawberries. Even at low concentrations, these pathogens can be a source of infection for consumers. Thus, their presence in strawberries is of particular significance as these are mostly consumed fresh and without any thermal processing. PRACTICAL APPLICATION: Nonlegislatively monitored pathogens of bacterial, viral and parasitic origin were found in strawberries. Monitoring the presence of these pathogens in ready-to-eat food is therefore meaningful and important in terms of food safety, especially in relation to pathogens with low infectious dose (for example, viruses, parasites).
- MeSH
- bezpečnost potravin MeSH
- Cryptosporidium izolace a purifikace MeSH
- Escherichia coli izolace a purifikace MeSH
- farmy MeSH
- Giardia lamblia izolace a purifikace MeSH
- jahodník mikrobiologie parazitologie virologie MeSH
- kontaminace potravin analýza MeSH
- lidé MeSH
- Listeria monocytogenes izolace a purifikace MeSH
- Norovirus izolace a purifikace MeSH
- počet mikrobiálních kolonií MeSH
- potravinářská mikrobiologie * MeSH
- potravinářská parazitologie MeSH
- průmyslově zpracované potraviny mikrobiologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Fresh vegetables and herbs are usually prepared and eaten raw without cooking or heating, which leads to a high risk of foodborne infection. The aim of the present study was to assess the contamination of raw vegetables, herbs, and the environment of food chains. Vegetable and herb samples originating both from the Czech Republic as well as from other countries were examined. The work was focused on the detection of commonly found, but also less frequently monitored foodborne pathogens, including viruses of the genus Norovirus (NoVs), hepatitis A virus (HAV), Listeria monocytogenes and Cronobacter spp. bacteria, and the parasites Cryptosporidium spp. and Giardia intestinalis. All samples were analyzed using individual RT-qPCR/qPCR assays; bacterial pathogens were also simultaneously detected using culture methods. The prevalence of the studied microorganisms in 623 samples ranged from 0.6% to 44.3% for individual pathogens. None of the samples were positive for the presence of HAV. Analysis of 157 environmental samples from 12 farms revealed the presence of NoVs in the environment of four farms. NoVs were detected in water samples as well as on the hands and gloves of workers. Escherichia coli was detected in all farms in the environmental samples and in eight farms in water samples. However, no sample of water exceeded the level of 100 CFU/mL for E. coli. None of the samples of water were positive for the presence of the studied parasites. Vegetables and herbs available from Czech markets and farms may pose a certain risk of foodborne disease, especially in the case of NoVs and parasites. PRACTICAL APPLICATION: This study provides valuable information on the microbiological quality of raw vegetables and herbs available from Czech markets and farms. Good hygienic practices aimed at reducing the incidence of pathogenic agents on fresh produce should not be neglected. Emphasis should be placed on the control of irrigation water, especially with respect to norovirus contamination. It is appropriate to combine culture methods and qPCR methods for the detection of bacterial agents.
- MeSH
- Cronobacter izolace a purifikace MeSH
- Cryptosporidium izolace a purifikace MeSH
- Escherichia coli izolace a purifikace MeSH
- farmy MeSH
- Giardia lamblia izolace a purifikace MeSH
- kontaminace potravin analýza MeSH
- kultivační média chemie MeSH
- kvalita jídla MeSH
- lidé MeSH
- Listeria monocytogenes izolace a purifikace MeSH
- Norovirus izolace a purifikace MeSH
- počet mikrobiálních kolonií MeSH
- potravinářská mikrobiologie MeSH
- potravinářská parazitologie MeSH
- virus hepatitidy A izolace a purifikace MeSH
- zelenina mikrobiologie parazitologie virologie MeSH
- zemědělci MeSH
- zemědělské plodiny mikrobiologie parazitologie virologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
Článek představuje méně obvyklou formu mimořádné události s hromadným postižením osob ve formě akutní gastroenteritidy, která je způsobená noroviry. Noroviry jsou malé neobalené RNA viry, které jsou u lidí častým původcem akutní gastroenteritidy. Vzplanutí hromadné nákazy je lehké z důvodu snadného přenosu, velmi nízké infekční dávky, krátké inkubační doby, snadného přežívání v prostředí a chybění trvalé imunity po překonaném onemocnění. Kazuistika představuje zásah poskytovatele zdravotnické záchranné služby v místě mimořádné události s hromadnou nákazou způsobenou noroviry. Jednalo se o školu v přírodě na území bývalého okresu Plzeň-sever, kde bylo stiženo akutní gastroenteritidou 56 dětí a 2 dospělé osoby. Popsána je reakce spádového zdravotnického operačního střediska, nasazování sil a prostředků zdravotnické složky, postup v místě události, organizace odsunu do zdravotnických zařízení a postup při příjmu nemocných dětí a jejich léčbě na Klinice infekčních nemocí a cestovní medicíny Fakultní nemocnice Plzeň.
The article presents a less common emergency situation with a mass casualty in the form of acute gastroenteritis caused by norovirus infection. Noroviruses are small non covered RNA viruses that are common source of acute gastroentritis. The spreading of mass infection is easy because of simple transmission, very low infective dose, short time of incubation, easy survival in the environment and lack of permanent immunity after overcoming the disease. The case report describes emergency medical service provider´s intervention on scene of multiple victim incident caused by norovirus infection. The incident was situated in the school camp in the former district of Pilsen – North, where 56 children and 2 adults suffered from acute gastroenteritis. The reaction of regional Medical Dispatch Center, deployed resources of Emergency Medical Service, on scene management and organization of transport to medical centers are described in the paper. The articela also deals with the procces of ill children´s admission and treatment at the Department of Infectious Diseases and Travel Medicine of the University Hospital and Medical School Plzeň.
Virové střevní infekce (akutní gastroenteritidy – AGE) jsou po respiračních infektech nejčastější příčinou infekčních onemocnění, a to celosvětově. Specifická prevence vakcinací je dostupná pouze u rotavirů, terapie je především symptomatická. Rozvoj molekulárně biologické diagnostiky, která zahrnuje všechny významné virové vyvolavatele gastroenteritid, vede k přesnějšímu monitoringu a následně i hlášení těchto infekcí.
Viral intestinal infections (acute gastroenteritides, AGE) are second only to respiratory infections as the most common cause of infectious disease worldwide. Specific prevention with vaccination is available only for rotaviruses; the treatment is mainly symptomatic. The development of molecular biology based diagnostic methods, involving all the major viral causative agents of gastroenteritides, leads to a more accurate monitoring and thus reporting of these infections.
- Klíčová slova
- virové gastroenteritidy,
- MeSH
- Adenoviridae patogenita MeSH
- adenovirové infekce lidí diagnóza patofyziologie terapie MeSH
- Astroviridae patogenita MeSH
- Coronaviridae patogenita MeSH
- gastroenteritida * diagnóza etiologie patofyziologie MeSH
- infekce viry z čeledi Astroviridae diagnóza patofyziologie terapie MeSH
- infekce viry z čeledi Coronaviridae diagnóza patofyziologie terapie MeSH
- lidé MeSH
- Norovirus patogenita MeSH
- rotavirové infekce diagnóza patofyziologie terapie MeSH
- Rotavirus patogenita MeSH
- Sapovirus patogenita MeSH
- vakcína proti rotavirům MeSH
- virové nemoci * diagnóza patofyziologie terapie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
- MeSH
- epidemie * statistika a číselné údaje MeSH
- Escherichia coli izolace a purifikace MeSH
- gastroenteritida * epidemiologie etiologie mikrobiologie MeSH
- lidé MeSH
- Norovirus izolace a purifikace MeSH
- pitná voda mikrobiologie škodlivé účinky MeSH
- vodní zdroje zásobování a distribuce MeSH
- zásobování vodou MeSH
- znečištění vody * MeSH
- Check Tag
- lidé MeSH
- Geografické názvy
- Česká republika MeSH
