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      Serological and Molecular Investigation of Batai Virus Infections in Ruminants from the State of Saxony-Anhalt, Germany, 2018

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          Abstract

          Arthropod-borne Batai virus (BATV) is an Orthobunyavirus widely distributed throughout European livestock and has, in the past, been linked to febrile diseases in humans. In Germany, BATV was found in mosquitoes and in one captive harbor seal, and antibodies were recently detected in various ruminant species. We have, therefore, conducted a follow-up study in ruminants from Saxony-Anhalt, the most affected region in Eastern Germany. A total of 325 blood samples from apparently healthy sheep, goats, and cattle were tested using a BATV-specific qRT-PCR and SNT. Even though viral RNA was not detected, the presence of antibodies was confirmed in the sera of all three species: sheep (16.5%), goats (18.3%), and cattle (41.4%). Sera were further analyzed by a glycoprotein Gc-based indirect ELISA to evaluate Gc-derived antibodies as a basis for a new serological test for BATV infections. Interestingly, the presence of neutralizing antibodies was not directly linked to the presence of BATV Gc antibodies. Overall, our results illustrate the high frequency of BATV infections in ruminants in Eastern Germany.

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          OptimalCutpoints: AnRPackage for Selecting Optimal Cutpoints in Diagnostic Tests

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            Orthobunyaviruses: recent genetic and structural insights.

            Orthobunyaviruses, which have small, tripartite, negative-sense RNA genomes and structurally simple virions composed of just four proteins, can have devastating effects on human health and well-being, either by causing disease in humans or by causing disease in livestock and crops. In this Review, I describe the recent genetic and structural advances that have revealed important insights into the composition of orthobunyavirus virions, viral transcription and replication and viral interactions with the host innate immune response. Lastly, I highlight outstanding questions and areas of future research.
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              A universal heterologous internal control system for duplex real-time RT-PCR assays used in a detection system for pestiviruses.

              A heterologous in vitro transcript based on a specific primer-probe HEX system was generated as a universal internal control (IC) to improve virus-specific real-time reverse-transcriptase PCR (RT-PCR) assays. By using a set of different primers, several PCR fragments of desired sizes of an in vitro transcript of the enhanced green fluorescent protein (EGFP) gene were generated, and the fragments were detected using a HEX-labelled probe. For long-term storage of the in vitro transcript a special RNA-safe buffer (RSB) was developed. Freezing and thawing of the IC diluted in RSB did not result in any substantial loss of detectable IC copy numbers. The new IC system was used for the first time in a duplex real-time RT-PCR assay for the detection of pestivirus-derived RNA, in particular from bovine viral diarrhea virus (BVDV). Primers and TaqMan probes for the 'panpesti' assay were selected by analysing the consensus sequence of the 5' non-translated region (5' NTR) of more than 600 different pestiviruses. Finally, the optimised primer probe combination showed an analytical sensitivity of less than 10 copies/reaction. In the duplex set-up, the analytical sensitivity of the validated real-time RT-PCR was identical to the sensitivity of the single assay without IC, and the diagnostic sensitivity of the duplex assay was equal or higher if compared to virus isolation.
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                Author and article information

                Contributors
                Role: Academic Editor
                Role: Academic Editor
                Journal
                Viruses
                Viruses
                viruses
                Viruses
                MDPI
                1999-4915
                26 February 2021
                March 2021
                : 13
                : 3
                : 370
                Affiliations
                [1 ]Institute of Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald-Insel Riems, Germany; nic.cichon@ 123456web.de (N.C.); martin.eiden@ 123456fli.de (M.E.); cora.holicki@ 123456fli.de (C.M.H.); martin.groschup@ 123456fli.de (M.H.G.)
                [2 ]Institute of Epidemiology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald-Insel Riems, Germany; jana.schulz@ 123456fli.de (J.S.); patrick.wysocki@ 123456fli.de (P.W.)
                [3 ]Institute of Diagnostic Virology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, 17493 Greifswald-Insel Riems, Germany; anne.guenther@ 123456fli.de
                [4 ]Department of Veterinary Medicine, State Office for Consumer Protection of Saxony-Anhalt, 39576 Stendal, Germany; joachim.borgwardt@ 123456sachsen-anhalt.de (J.B.); wolfgang.gaede@ 123456sachsen-anhalt.de (W.G.)
                Author notes
                [* ]Correspondence: ute.ziegler@ 123456fli.de
                Author information
                https://orcid.org/0000-0002-1197-8288
                https://orcid.org/0000-0002-5719-4542
                https://orcid.org/0000-0002-1258-5646
                https://orcid.org/0000-0003-0215-185X
                https://orcid.org/0000-0001-5295-7339
                Article
                viruses-13-00370
                10.3390/v13030370
                7996813
                33652882
                39ccb767-06c8-4870-99c9-4c6b4dbdbf89
                © 2021 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 15 January 2021
                : 23 February 2021
                Categories
                Article

                Microbiology & Virology
                batai virus,elisa,seroprevalence,ruminants,germany
                Microbiology & Virology
                batai virus, elisa, seroprevalence, ruminants, germany

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