HAV biomarker 2A can be used to differentiate between previously HAV-vaccinated and naturally infected individuals. Within a multiplex serological approach this assay can provide valuable novel information in the context of outbreak investigations, longitudinal population based studies and evaluations of immunization campaigns.
Caveolin expression supports the multiplication of retro-, ortho- and paramyxoviruses in susceptible cells. However, human influenza A virus (IAV), an orthomyxovirus, does not multiply efficiently in mouse embryo fibroblasts (MEFs), which are abundant in caveolin-1 (Cav-1). Surprisingly, the absence of Cav-1 in a MEF cell line removed the block for IAV replication and raised the infectious titer 250-fold, whereas the re-introduction of Cav-1 reversed the effect. The monitoring of cellular pathways revealed that Cav-1 loss considerably increased activities of p53. Furthermore, infection of MEF Cav-1 (-/-) induced reactive oxygen species (ROS) and pronounced apoptosis in the late phase of viral multiplication, but no type I IFN response. Strikingly, pharmacological inactivation showed that the elevated levels of ROS together with apoptosis caused the increase of virus yield. Thus, Cav-1 represents a new negative regulator of IAV infection in MEF that diminishes IAV infectious titer by controlling virus-supportive pathways.
Hepatitis E, a liver disease caused by infection with the hepatitis E virus (HEV), is a worldwide emerging disease. The diagnosis is based on the detection of viral RNA and of HEV-specific immunoglobulins (Ig). For the latter, various assays are commercially available but still lack harmonization. In this study, a Luminex-based multiplex serological assay was established that measures the presence of total IgG, IgA, and IgM antibodies, targeting a short peptide derived from the viral E2 protein. For the validation, 160 serum samples with a known HEV serostatus were used to determine the assay cutoff and accuracy. Thereby, HEV IgG- and RNA-positive sera were identified with a sensitivity of 100% and a specificity of 98% (95% confidence interval [CI], 94% to 100%). Application of the assay by retesting 514 serum samples previously characterized with different HEV-IgG or total antibody tests revealed a high level of agreement between the assays (Cohen’s kappa, 0.58 to 0.99). The established method is highly sensitive and specific and can be easily implemented in a multiplex format to facilitate rapid differential diagnostics with a few microliters of sample input.
Purpose:The discovery of Lloviu virus (LLOV) in Miniopterus schreibersii bat samples from Spain, 2003 dramatically changed our understanding of the genetic diversity, geographic distribution, and host preference of filoviruses. However, the ecology of LLOV is largely unclear mainly due to the lack of reports following the first report of LLOV. In our study, we try to solve the open questions regarding LLOV genetics and ecology, based on a continuous screening of a selected M. schreibersii colony, proved to be positive for LLOV during a mortality event in 2016. Methods & Materials:We established a countrywide surveillance system in Hungary for the early detection of M. schreibersii dye-offs in 2012 in collaboration with conservation bilogists and chiropterologists. In each case, carcasses were collected as soon as possible and transported to the laboratory in liquid nitrogen. Multiple events were examined during the past few years with viral metagenomic analyses and LLOV-specific TaqMan-based real-time PCR screening.Results: In 2016, we detected LLOV virus RNA in tissue samples of a M. schreibersii individual. Partial sequences of the nucleoprotein and the RNA-dependent RNA-polymerase gene suggests a close genetic relatedness with the original isolate in Spain. Following this event, several additional mortalities were registered to date in the same habitat with the same gross pathology of hemorrhagic symptoms, but no other positives were verified with PCR method, possibly because of the bad conditioned carcasses. In 2018, we started a monthly sampling activity, after the maternity period, in order to examine the seroprevalence, and other related factors of the virus in this cave.Conclusion: Here we present the current results of our survey programme, showing the relation of the Hungarian isolate to the original Spanish virus from 2003. A major goal of our presentation is to call attention to this pathogen, possibly affecting the stability of M. schreibersii colonies across Europe, representing a paramount concern for conservation biology. We discuss the possible factors leading to the dispersal of the virus in Europe and the possible transmission routes between bats to be examined in future studies and we also summarize current knowledge about the virus.Purpose: The alphavirus genus comprises several human pathogens, e.g. chikungunya virus (CHIKV), Ross River virus (RRV) and Eastern equine encephalitis virus (EEEV). Depending on the antigenic relatedness cross-reactivity in serological assays is commonly observed. Due to overlapping geographical distribution and similar clinical pictures a serological differentiation is desirable.Methods & Materials: Human serum or plasma samples from different geographical areas and precharacterised for IgG and IgM antibodies against various alphaviruses were investigated with multiplex indirect immunofluorescence assays. The method is based on a mosaic of up to nine biochips which carry infected or transfected cell lines each expressing antigens of another alphavirus. The sample...
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