Human West Nile Virus Research Articles

Contrasted Epidemiological Patterns of West Nile Virus Lineages 1 and 2 Infections in France from 2015 to 2019.

Since 2015, annual West Nile virus (WNV) outbreaks of varying intensities have been reported in France. Recent intensification of enzootic WNV circulation was observed in the South of France with most horse cases detected in 2015 (n = 49), 2018 (n = 13), and 2019 (n = 13). A WNV lineage 1 strain was isolated from a horse suffering from West Nile neuro-invasive disease (WNND) during the 2015 episode in the Camargue area. A breaking point in WNV epidemiology was achieved in 2018, when WNV lineage 2 emerged in Southeastern areas. This virus most probably originated from WNV spread from Northern Italy and caused WNND in humans and the death of diurnal raptors. WNV lineage 2 emergence was associated with the most important human WNV epidemics identified so far in France (n = 26, including seven WNND cases and two infections in blood and organ donors). Two other major findings were the detection of WNV in areas with no or limited history of WNV circulation (Alpes-Maritimes in 2018, Corsica in 2018–2019, and Var in 2019) and distinct spatial distribution of human and horse WNV cases. These new data reinforce the necessity to enhance French WNV surveillance to better anticipate future WNV epidemics and epizootics and to improve the safety of blood and organ donations.

The development of a spatially resolved retrospective ensemble forecast model of West Nile virus in Long Island, New York

The development of a spatially resolved retrospective ensemble forecast model of West Nile virus in Long Island, New York Nicholas DeFelice, Meytar Sorek-Hamer, Scott R Campbell and Krishna Vemuri West Nile virus (WNV) emerged in the western hemisphere in 1999, and has established itself as the leading cause of domestically acquired arboviral disease in the United States. Though the transmission of WNV exhibits a pronounced sensitivity to a complex seasonal ecology along with hydrological and meteorological conditions, our ability to predict the timing, duration, and magnitude of local WNV outbreaks remains limited. In Long Island, New York, annual peak estimates of WNV infected mosquitos have ranged from 1 in 300 to 1 in 50 resulting in anywhere from 0 to 24 reported human WNV cases, respectively. Here we report the development of a spatially refined model that uses ECOsystem Spaceborne Thermal Radiometer Experiment on Space Station (ECOSTRESS) data to capture the variability in physical environmental factors (e.g., temperature and hydrology) along with a compartmental model describing WNV transmission dynamics, to retrospectively forecast WNV outbreaks in Long Island, New York. The inclusion of ECOSTRESS’ high spatial (70m) and temporal (revisits every 4 days) resolution data will allow us to capture changes in the micro-ecosystem. Ensemble simulations with the WNV model are iteratively optimized using data assimilation methods and observations of human incidence, mosquito abundance, and mosquito infection rates. The ECOSTRESS meteorological and hydrological indicators are included in the core model structure to better constrain WNV amplification and transmission dynamics. The model-inference system can be used to better understand the spatial variability of the outbreak and estimate the relationship between zoonotic amplification and human outbreaks. This work represents an initial step in the development of a statistically rigorous system for a spatially resolved real-time forecast of seasonal outbreaks of West Nile virus.

Transmission of West Nile and five other temperate mosquito-borne viruses peaks at temperatures between 23°C and 26°C.

The temperature-dependence of many important mosquito-borne diseases has never been quantified. These relationships are critical for understanding current distributions and predicting future shifts from climate change. We used trait-based models to characterize temperature-dependent transmission of 10 vector-pathogen pairs of mosquitoes (Culex pipiens, Cx. quinquefascsiatus, Cx. tarsalis, and others) and viruses (West Nile, Eastern and Western Equine Encephalitis, St. Louis Encephalitis, Sindbis, and Rift Valley Fever viruses), most with substantial transmission in temperate regions. Transmission is optimized at intermediate temperatures (23-26°C) and often has wider thermal breadths (due to cooler lower thermal limits) compared to pathogens with predominately tropical distributions (in previous studies). The incidence of human West Nile virus cases across US counties responded unimodally to average summer temperature and peaked at 24°C, matching model-predicted optima (24-25°C). Climate warming will likely shift transmission of these diseases, increasing it in cooler locations while decreasing it in warmer locations.

An Agent-Based Model of West Nile Virus: Predicting the Impact of Public Health Agents and Vaccinations on Horses

West Nile Virus (WNV), primarily spread by the Culex species of mosquito, accounts for a large percentage of mosquito-borne diseases. In order to investigate the dynamics of WNV, an agent-based model was developed in the NetLogo modeling environment that simulates the transmission of the virus by incorporating temperature, humans, horses, birds, and a mosquito population. This model allows the observer to view simulations between the agents listed and measure the impact of vaccinations on the survival rate of horses infected with WNV. Furthermore, the model integrates and evaluates the impact public health agents have on human WNV dynamics through their education of the general public and containment of mosquito populations. The intention of the simulation results from this model is to help develop a vaccination strategy for horse populations, while also demonstrating the usefulness of public health agents’ efforts on the individual agents within the disease dynamics.

West Nile virus in humans, Greece, 2018: the largest seasonal number of cases, 9 years after its emergence in the country.

BackgroundHuman cases of West Nile virus (WNV) infection are recorded since 2010 in Greece, with seasonal outbreaks occurring almost annually. Enhanced surveillance has been implemented since 2010, to promptly characterise cases’ temporal and geographical distribution and inform authorities for implementation of appropriate measures (mosquito control, health education, blood safety).AimWe describe the epidemiology of WNV human infections in Greece focusing on the 2018 season.MethodsThe National Public Health Organization advised physicians to test all suspect WNV infection cases and refer samples to reference laboratories. Laboratories notified diagnosed cases on a daily basis. Treating physicians, patients, and infected blood donors were interviewed within 48 hours after diagnosis and the probable infection location was identified. Hospitalised cases were followed up until discharge.ResultsA total of 317 autochthonous WNV infection cases were diagnosed in 2018. Among them, 243 cases had neuroinvasive disease (WNND), representing a 23% increase of WNND cases compared with 2010, the previous most intense season. There were 51 deaths. Cases started occurring from week 22, earlier than usual. Both rural and urban areas were affected, with 86 (26% of the total) municipalities belonging to seven (54% of the total) regions recording cases. Two major epicentres were identified in Attica and Central Macedonia regions.ConclusionsThe largest number of human cases of WNV infection ever recorded in Greece occurred in 2018, with a wide geographical distribution, suggesting intense virus circulation. Enhanced surveillance is vital for the early detection of human cases and the prompt implementation of response measures.

Detection of Japanese Encephalitis Virus in Culex Mosquitoes in Singapore.

.Mosquito-borne flaviviruses are emerging pathogens of an increasing global public health concern because of their rapid increase in geographical range and the impact of climate change. Japanese encephalitis virus (JEV) and West Nile virus (WNV) are of concern because of the risk of reemergence and introduction by migratory birds. In Singapore, human WNV infection has never been reported and human JEV infection is rare. Four sentinel vector surveillance sites were established in Singapore to understand the potential risk posed by these viruses. Surveillance was carried out from August 2011 to December 2012 at Pulau Ubin, from March 2011 to March 2013 at an Avian Sanctuary (AS), from December 2010 from October 2012 at Murai Farmway, and from December 2010 to December 2013 at a nature reserve. The present study revealed active JEV transmission in Singapore through the detection of JEV genotype II in Culex tritaeniorhynchus collected from an Avian Sanctuary. Culex flavivirus (CxFV), similar to the Quang Binh virus isolated from Cx. tritaeniorhynchus in Vietnam and CxFV-LSFlaviV-A20-09 virus isolated in China, was also detected in Culex spp. (vishnui subgroup). No WNV was detected. This study demonstrates the important role that surveillance plays in public health and strongly suggests the circulation of JEV among wildlife in Singapore, despite the absence of reported human cases. A One Health approach involving surveillance, the collaboration between public health and wildlife managers, and control of mosquito populations remains the key measures in risk mitigation of JEV transmission in the enzootic cycle between birds and mosquitoes.

West Nile Virus in Slovenia.

West Nile virus (WNV) is a flavivirus transmitted by mosquitoes. Birds are the reservoir for the virus; humans, horses and other mammals are dead-end hosts. Infections caused by WNV in humans can vary from asymptomatic infections to West Nile fever (WNF) or West Nile neuroinvasive disease (WNND). In 1995, a serosurvey was performed in Slovenia on forest workers, and WNV specific IgG antibodies were confirmed in 6.8% of the screened samples, indicating that WNV is circulating in Slovenia. No human disease cases were detected in Slovenia until 2013, when the first case of WNV infection was confirmed in a retrospective study in a 79-year old man with meningitis. In 2018, three patients with WNND were confirmed by laboratory tests, with detection of IgM antibodies in the cerebrospinal fluid of the patients. In one of the patients, WNV RNA was detected in the urine sample. In 2017, 2018 and 2019, a mosquito study was performed in Slovenia. Mosquitoes were sampled on 14 control locations and 35 additional locations in 2019. No WNV was detected in mosquitoes in 2017 and 2019, but we confirmed the virus in a pool of Culex sp. mosquitoes in 2018. The virus was successfully isolated, and complete genome sequence was acquired. The whole genome of the WNV was also sequenced from the patient’s urine sample. The whole genome sequences of the WNV virus detected in Slovenian patient and mosquito indicate the virus most likely spread from the north, because of the geographic proximity and because the sequences cluster with the Austrian and Hungarian sequences. A sentinel study was performed on dog sera samples, and we were able to confirm IgG antibodies in 1.8% and 4.3% of the samples in 2017 and 2018, respectively. Though Slovenia is not a highly endemic country for WNV, we have established that the virus circulates in Slovenia.

Temporal and Spatial Impacts of Hurricane Damage on West Nile Virus Transmission and Human Risk.

Hurricanes have profound impacts on zoonotic pathogen ecosystems that exhibit spatial and temporal waves in both distance from and time since the event. Wind, rain, and storm surge directly affect mosquito vectors and animal hosts of these pathogens. In this analysis, we apply a West Nile virus transmission model parameterized for the Northern coast of the Gulf of Mexico to explore the effect of event timing of hurricane landfall, time since the event, and damage extent on human West Nile virus neuro-invasive disease (WNV-NID) risk. Early-season hurricanes, which make landfall prior to the peak of baseline WNV transmission activity, increase the average total WNV-infectious mosquitoes for the year by 7.8% and human WNV-NID incidence by 94.3% across all areas with hurricane damage. The indirect effects on human exposure to mosquito bites in the immediate aftermath and long-term recovery from the event have strong impacts on the risk of infection. The resultant interactive direct and indirect storm effects on the pathogen system are spatially and temporally heterogenous among the generalized time and space categories modeled.

The drivers of West Nile virus human illness in the Chicago, Illinois, USA area: Fine scale dynamic effects of weather, mosquito infection, social, and biological conditions.

West Nile virus (WNV) has consistently been reported to be associated with human cases of illness in the region near Chicago, Illinois. However, the number of reported cases of human illness varies across years, with intermittent outbreaks. Several dynamic factors, including temperature, rainfall, and infection status of vector mosquito populations, are responsible for much of these observed variations. However, local landscape structure and human demographic characteristics also play a key role. The geographic and temporal scales used to analyze such complex data affect the observed associations. Here, we used spatial and statistical modeling approaches to investigate the factors that drive the outcome of WNV human illness on fine temporal and spatial scales. Our approach included multi-level modeling of long-term weekly data from 2005 to 2016, with weekly measures of mosquito infection, human illness and weather combined with more stable landscape and demographic factors on the geographical scale of 1000m hexagons. We found that hot weather conditions, warm winters, and higher MIR in earlier weeks increased the probability of an area of having a WNV human case. Higher population and the proportion of urban light intensity in an area also increased the probability of observing a WNV human case. A higher proportion of open water sources, percentage of grass land, deciduous forests, and housing built post 1990 decreased the probability of having a WNV case. Additionally, we found that cumulative positive mosquito pools up to 31 weeks can strongly predict the total annual human WNV cases in the Chicago region. This study helped us to improve our understanding of the fine-scale drivers of spatiotemporal variability of human WNV cases.

West Nile Virus Lineage 2 Vector Competence of Indigenous Culex and Aedes Mosquitoes from Germany at Temperate Climate Conditions.

West Nile virus (WNV) is a widespread zoonotic arbovirus and a threat to public health in Germany since its first emergence in 2018. It has become of particular relevance in Germany in 2019 due to its rapid geographical spread and the detection of the first human clinical cases. The susceptibility of indigenous Culex pipiens (biotypes pipiens and molestus) for a German WNV lineage 2 strain was experimentally compared to that of Serbian Cx. pipiens biotype molestus and invasive German Aedes albopictus. All tested populations proved to be competent laboratory vectors of WNV. Culex pipiens biotype pipiens displayed the highest transmission efficiencies (40.0%–52.9%) at 25 °C. This biotype was also able to transmit WNV at 18 °C (transmission efficiencies of 4.4%–8.3%), proving that temperate climates in Central and Northern Europe may support WNV circulation. Furthermore, due to their feeding behaviors, Cx. pipiens biotype molestus and Ae. albopictus can act as “bridge vectors”, leading to human WNV infections.

West Nile Virus Epidemic in Germany Triggered by Epizootic Emergence, 2019.

One year after the first autochthonous transmission of West Nile virus (WNV) to birds and horses in Germany, an epizootic emergence of WNV was again observed in 2019. The number of infected birds and horses was considerably higher compared to 2018 (12 birds, two horses), resulting in the observation of the first WNV epidemy in Germany: 76 cases in birds, 36 in horses and five confirmed mosquito-borne, autochthonous human cases. We demonstrated that Germany experienced several WNV introduction events and that strains of a distinct group (Eastern German WNV clade), which was introduced to Germany as a single introduction event, dominated mosquito, birds, horse and human-related virus variants in 2018 and 2019. Virus strains in this clade are characterized by a specific-Lys2114Arg mutation, which might lead to an increase in viral fitness. Extraordinary high temperatures in 2018/2019 allowed a low extrinsic incubation period (EIP), which drove the epizootic emergence and, in the end, most likely triggered the 2019 epidemic. Spatiotemporal EIP values correlated with the geographical WNV incidence. This study highlights the risk of a further spread in Germany in the next years with additional human WNV infections. Thus, surveillance of birds is essential to provide an early epidemic warning and thus, initiate targeted control measures.

Evaluation of a West Nile virus risk-assessment tool used at a local health unit

In Ontario, public health units collect surveillance data on vector-borne diseases (VBD) to determine emerging trends and develop VBD management strategies. Risk-assessment tools that are simple and easily applied can provide public health practitioners with objective evaluations of the risk of West Nile virus (WNV) activity in their jurisdiction. This study was conducted to evaluate an existing WNV risk-assessment tool used by a public health unit in southern Ontario. The purpose of this study was to: (i) describe the trends for WNV in mosquito and human cases in the Region of Peel, Ontario, Canada, and (ii) investigate the ability of the risk-assessment tool to predict positive human cases and positive mosquito traps in the following weeks. Data were collected from 2011 to 2016 and analysed using simple descriptive statistics and Fisher’s exact tests. This study found the tool includes variables that are not significant in predicting WNV activity in the following weeks. The current tool should be revised to remove variables that are not significant in predicting risk and add additional variables that have been shown to be effective predictors in other studies, such as rainfall and human WNV cases in the previous year.

Can local risk of West Nile virus infection be predicted from previous cases? A descriptive study in Quebec, 2011-2016.

This study aimed at (1) describing the local risk of West Nile virus (WNV) infection in humans based on previous case reports and (2) investigating the spatial clustering of cases in the five most affected administrative regions of Quebec, Canada, for the 2011-2016 period. Human WNV cases declared to the Ministry of Health and Social Services of Quebec(Ministère de la santé et des services sociaux, MSSS) were retrieved. Incidence risk by age and sex was calculated for the study period. The yearly and monthly occurrence of cases in geographical units (GUs) was described and the probability of observing cases in a GU with cases reported in the previous year or month was assessed. Moran's I was used to assess global clustering across the study area. Spatial clusters were identified by the Kulldorff scan statistic. A total of 261 WNV cases were declared to the MSSS between 2011 and 2016 in the study area. Overall, a low percentage of GU with cases reported had additional cases reported over the next month or year. Global spatial clustering was weak but statistically significant (p < 0.05) for 2012 and 2015. For these two years, spatial clusters of high-risk GUs were identified. Results underline the challenge of predicting the distribution of WNV incidence risk in Quebec based on previous occurrence of human cases. Ongoing research with high spatial resolution entomological data is still necessary to understand the spatial distribution of risk at a local scale.

Comparative Pathology of West Nile Virus in Humans and Non-Human Animals.

West Nile virus (WNV) continues to be a major cause of human arboviral neuroinvasive disease. Susceptible non-human vertebrates are particularly diverse, ranging from commonly affected birds and horses to less commonly affected species such as alligators. This review summarizes the pathology caused by West Nile virus during natural infections of humans and non-human animals. While the most well-known findings in human infection involve the central nervous system, WNV can also cause significant lesions in the heart, kidneys and eyes. Time has also revealed chronic neurologic sequelae related to prior human WNV infection. Similarly, neurologic disease is a prominent manifestation of WNV infection in most non-human non-host animals. However, in some avian species, which serve as the vertebrate host for WNV maintenance in nature, severe systemic disease can occur, with neurologic, cardiac, intestinal and renal injury leading to death. The pathology seen in experimental animal models of West Nile virus infection and knowledge gains on viral pathogenesis derived from these animal models are also briefly discussed. A gap in the current literature exists regarding the relationship between the neurotropic nature of WNV in vertebrates, virus propagation and transmission in nature. This and other knowledge gaps, and future directions for research into WNV pathology, are addressed.

The European Union One Health 2018 Zoonoses Report.

This report of the European Food Safety Authority and the European Centre for Disease Prevention and Control presents the results of zoonoses monitoring activities carried out in 2018 in 36 European countries (28 Member States (MS) and 8 non‐MS). The first and second most commonly reported zoonoses in humans were campylobacteriosis and salmonellosis, respectively. The European Union (EU) trend for confirmed human cases of these two diseases was stable during 2014–2018. The proportion of human salmonellosis cases due to Salmonella Enteritidis was at the same level in 2018 as in 2017. Of the 27 reporting MS, 16 met all Salmonella reduction targets for poultry, whereas 11 MS failed meeting at least one. The EU flock prevalence of target Salmonella serovars in breeding hens, laying hens, broilers and fattening turkeys decreased during recent years but stalled in breeding turkeys. Salmonella results from Competent Authorities for pig carcasses and for poultry tested through National Control Programmes were more frequently positive compared with food business operators. Shiga toxin‐producing Escherichia coli (STEC) infections in humans were the third most commonly reported zoonosis in the EU and increased from 2014 to 2018. Yersiniosis was the fourth most frequently reported zoonosis in humans in 2018 with a stable trend in 2014–2018. The number of reported confirmed listeriosis cases further increased in 2018, despite Listeria rarely exceeding the EU food safety limit tested in ready‐to‐eat food. In total, 5,146 food‐ and waterborne outbreaks were reported. Salmonella was the most commonly detected agent with S. Enteritidis causing one in five outbreaks. Salmonella in eggs and egg products was the highest risk agent/food pair. A large increase of human West Nile virus infections was reported in 2018. The report further updates on bovine tuberculosis, Brucella, Trichinella, Echinococcus, Toxoplasma, rabies, Coxiella burnetii (Q fever) and tularaemia.

West Nile Virus Infection Blocks Inflammatory Response and T Cell Costimulatory Capacity of Human Monocyte-Derived Dendritic Cells.

West Nile virus (WNV) is a neurotropic flavivirus and the leading cause of mosquito-borne encephalitis in the United States. Recent studies in humans have found that dysfunctional T cell responses strongly correlate with development of severe WNV neuroinvasive disease. However, the contributions of human dendritic cells (DCs) in priming WNV-specific T cell immunity remains poorly understood. Here, we demonstrate that human monocyte derived DCs (moDCs) support productive viral replication following infection with a pathogenic strain of WNV. Antiviral effector gene transcription was strongly induced during the log phase of viral growth, while secretion of type I interferons (IFN) occurred with delayed kinetics. Activation of RIG-I like receptor (RLR) or type I IFN signaling prior to log phase viral growth significantly diminished viral replication, suggesting that early activation of antiviral programs can block WNV infection. In contrast to the induction of antiviral responses, WNV infection did not promote transcription or secretion of proinflammatory (interleukin-6 [IL-6], granulocyte-macrophage colony-stimulating factor [GM-CSF], CCL3, CCL5, and CXCL9) or T cell modulatory (IL-4, IL-12, and IL-15) cytokines. There was also minimal induction of molecules associated with antigen presentation and T cell priming, including the costimulatory molecules CD80, CD86, and CD40. Functionally, WNV-infected moDCs dampened allogenic CD4 and CD8 T cell activation and proliferation. Combining these observations, we propose a model whereby WNV subverts human DC activation to compromise priming of WNV-specific T cell immunity.IMPORTANCE West Nile virus (WNV) is an encephalitic flavivirus that remains endemic in the United States. Previous studies have found dysfunctional T cell responses correlate to severe disease outcomes during human WNV infection. Here, we sought to better understand the ability of WNV to program human dendritic cells (DCs) to prime WNV-specific T cell responses. While productive infection of monocyte-derived DCs activated antiviral and type I interferon responses, molecules associated with inflammation and programming of T cells were minimally induced. Functionally, WNV-infected DCs dampened T cell activation and proliferation during an allogeneic response. Combined, our data support a model whereby WNV infection of human DCs compromises WNV-specific T cell immunity.

Environmental and Sociological Factors Associated with the Incidence of West Nile Virus Cases in the Northern San Joaquin Valley of California, 2011-2015.

Environmental and socioeconomic risk factors associated with the incidence of human West Nile virus (WNV) cases were investigated in the Northern San Joaquin Valley region of California, a largely rural area. The study included human WNV cases from the years 2011 to 2015 in the three-county area of San Joaquin, Stanislaus, and Merced Counties, and used census tracts as the unit of analysis. Environmental factors included temperature, precipitation, and WNV-positive mosquito pools. Socioeconomic variables included age, housing age, housing foreclosures, median income, and ethnicity. Chi-square independence tests were used to examine whether each variable was associated with the incidence of WNV cases using data from the three counties combined. In addition, negative binomial regression revealed that the environmental factors of temperature and precipitation were the strongest predictors of the incidence of human WNV cases, while the socioeconomic factor of ethnicity was a significant predictor as well, and is a factor to consider in prevention efforts. Source reduction of mosquito breeding sites and targeted prevention and education remain key in reducing the risk associated with WNV.

Reducing West Nile Virus Risk Through Vector Management.

Over 50,000 human West Nile virus (WNV) (Flaviviridae: Flavivirus) clinical disease cases have been reported to the CDC during the 20 yr that the virus has been present in the United States. Despite the establishment and expansion of WNV-focused mosquito surveillance and control efforts and a renewed emphasis on applying integrated pest management (IPM) principles to WNV control, periodic local and regional WNV epidemics with case reports exceeding 2,000 cases per year have occurred during 13 of those 20 yr in the United States. In this article, we examine the scientific literature for evidence that mosquito control activities directed at either preventing WNV outbreaks or stopping those outbreaks once in progress reduce WNV human disease or have a measurable impact on entomological indicators of human WNV risk. We found that, despite a proliferation of research investigating larval and adult mosquito control effectiveness, few of these studies actually measure epidemiological outcomes or the entomological surrogates of WNV risk. Although many IPM principles (e.g., control decisions based on surveillance, use of multiple control methodologies appropriate for the ecosystem) have been implemented effectively, the use of action thresholds or meaningful public health outcome assessments have not been used routinely. Establishing thresholds for entomological indicators of human risk analogous to the economic injury level and economic thresholds utilized in crop IPM programs may result in more effective WNV prevention.

Antibiotic Minocycline Prevents Respiratory Syncytial Virus Infection.

Treatment drugs, besides their specific activity, often have multiple effects on the body. The undesired effect of the drug may be repurposed as therapeutics, saving significant investigative time and effort. Minocycline has anti-cancer, anti-oxidant, anti-inflammatory, and anti-apoptotic properties. Presently, minocycline is also known to show anti-viral activity against Influenza virus, Japanese encephalitis virus, Simian immunodeficiency virus, Human immunodeficiency virus and West Nile virus. Here, we investigate the effect of minocycline on Respiratory syncytial virus (RSV), a common respiratory virus that causes severe mortality and morbidity in infants, children, and older adult populations. Currently, there is no effective vaccine or treatment for RSV infection; hence, there is a critical need for alternative and effective drug choices. Our study shows that minocycline reduces the RSV-mediated cytopathic effect and prevents RSV infection. This is the first study demonstrating the anti-viral activity of minocycline against RSV.

Extraordinary increase in West Nile virus cases and first confirmed human Usutu virus infection in Hungary, 2018.

BackgroundDuring the 2018 WNV transmission season, similarly to other endemic areas in Europe, a large number of human West Nile virus (WNV) infections were reported in Hungary.AimsWe summarise the epidemiological and laboratory findings of the 2018 transmission season and expand experiences in flavivirus differential diagnostics.MethodsEvery patient with clinical suspicion of acute WNV infection was in parallel tested for WNV, tick-borne encephalitis virus and Usutu virus (USUV) by serological methods. Sera, whole blood and urine samples were also tested for the presence of viral nucleic acid.ResultsUntil the end of December 2018, 215 locally acquired and 10 imported human WNV infections were notified in Hungary. All reported cases were symptomatic; most of them exhibited neurological symptoms. In a large proportion of tested individuals, whole blood was the most appropriate sample type for viral nucleic acid detection, but because whole blood samples were not always available, testing of urine samples also extended diagnostic possibilities. In addition, the first human USUV infection was confirmed in 2018 in a patient with aseptic meningitis. Serological cross-reactions with WNV in different serological assays were experienced, but subsequent molecular biological testing and sequence analysis identified Europe lineage 2 USUV infection.ConclusionCareful interpretation and simultaneous application of different laboratory methods are necessary to avoid misdiagnosis of human USUV cases. Expansion of the laboratory-confirmed case definition criteria for detection of viral RNA in any clinical specimens to include urine samples could increase diagnostic sensitivity.