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Surveillance and Pathogenicity of Lyssaviruses

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Surveillance and Pathogenicity of Lyssaviruses Inaugural-Dissertation zur Erlangung der Doktorwürde der Tierärztlichen Fakultät der Ludwig-Maximilians-Universität München Surveillance and pathogenicity of lyssaviruses von Elisa Dorothea Eggerbauer aus Köln München 2018 Aus dem Veterinärwissenschaftlichen Department der Tierärztlichen Fakultät der Ludwig-Maximilians-Universität München Lehrstuhl für Virologie Arbeit angefertigt unter der Leitung von Univ.-Prof. Dr. Gerd Sutter Angefertigt am Institut für molekulare Virologie und Zellbiologie, Friedrich-Loeffler-Institut, Bundesforschungsinstitut für Tiergesundheit, Insel Riems Mentoren: Prof. Dr. Dr. h.c. Thomas C. Mettenleiter & Prof. Dr. Martin G. Beer Gedruckt mit der Genehmigung der Tierärztlichen Fakultät der Ludwig-Maximilians-Universität München Dekan: Univ.-Prof. Dr. Reinhard K. Straubinger Ph.D Berichterstatter: Univ.-Prof. Dr. Gerd Sutter Korreferent/en: Univ.-Prof. Dr. Katrin Hartmann Tag der Promotion: 10. Februar 2018 Die vorliegende Arbeit wurde gemäß § 6 Abs. 2 der Promotionsordnung für die Tierärztliche Fakultät der Ludwig-Maximilians-Universität München in kumulativer Form verfasst. Folgende wissenschaftliche Arbeiten sind in dieser Dissertationsschrift enthalten: Eggerbauer, E., Pfaff, F., Finke, S., Höper, D., Beer, M., Mettenleiter, T.C., Nolden, T., Teifke, J.P., Müller, T., Freuling, C.M. Comparative analysis of European bat lyssavirus 1 pathogenicity in the mouse model. PLOS Neglected Tropical Diseases 2017; 11(6): e0005668. https://doi.org/10.1371/journal.pntd.0005668 Eggerbauer, E., Troupin, C., Passior, K., Pfaff, F., Höper, D., Neubauer-Juric, A., Haberl, S., Bouchier, C., Mettenleiter, T.C., Bourhy, H., Müller, T., Dacheux, L., Freuling, C.M. 2017. The recently discovered Bokeloh bat lyssavirus: Insights into its genetic heterogeneity and spatial distribution in Europe and the population genetics of its primary host. In Loeffler’s Footsteps – Viral Genomics in the Era of High-Throughput Sequencing. Advances in Virus Research, 2017. https://doi.org/10.1016/bs.aivir.2017.07.004 (in press) Eggerbauer, E., de Benedictis, P., Hoffmann, B., Mettenleiter, T.C., Schlottau, K., Ngoepe, E.C., Sabeta, C.T., Freuling, C.M., Müller, T. Evaluation of Six Commercially Available Rapid Immunochromatographic Tests for the Diagnosis of Rabies in Brain Material. PLOS Neglected Tropical Diseases 2016; 10(6): e0004776. https://doi.org/10.1371/journal.pntd.0004776 For my family Table of contents 1. Abbreviations ......................................................................................................................... 1 2. Introduction ............................................................................................................................ 5 3. Literature review .................................................................................................................... 7 3.1. History of rabies with focus on Europe ...................................................................... 7 3.2. Lyssaviruses ................................................................................................................ 8 3.2.1. Virus Taxonomy .............................................................................................................. 8 3.2.2. Virus structure and viral genome.................................................................................. 11 3.2.2.1. Viral RNA ..................................................................................................... 12 3.2.2.2. Viral proteins ............................................................................................... 12 3.2.3. Replication cycle ........................................................................................................... 14 3.3. Rabies disease .......................................................................................................... 15 3.3.1. Pathogenesis................................................................................................................. 16 3.3.2. Clinical picture .............................................................................................................. 16 3.3.3. Pathogenicity ................................................................................................................ 17 3.3.4. Vaccination and post exposure prophylaxis ................................................................. 18 3.4. Rabies epidemiology ................................................................................................ 19 3.4.1. Terrestrial rabies ........................................................................................................... 19 3.4.2. Bat rabies ...................................................................................................................... 20 3.4.2.1. Bat rabies in Europe .................................................................................... 21 3.4.2.2. Passive bat rabies surveillance in Germany ................................................ 23 3.5. Rabies diagnosis ....................................................................................................... 25 3.5.1. Standard diagnostic tests.............................................................................................. 25 3.5.2. Challenges in rabies diagnosis ...................................................................................... 26 3.5.3. Alternative diagnostic tests .......................................................................................... 27 4. Objectives ............................................................................................................................. 31 5. Results .................................................................................................................................. 33 I 5.1. Comparative analysis of European bat lyssavirus 1 isolates in the mouse model... 33 5.2. The recently discovered Bokeloh bat lyssavirus – Insights into its genetic heterogeneity and spatial distribution in Europe and the population genetics of its primary host ............................................................................................................. 65 5.3. Evaluation of six commercially available rapid immunochromatographic tests for the diagnosis of rabies in brain material ............................................................... 109 6. Discussion ........................................................................................................................... 133 7. Summary............................................................................................................................. 143 8. Zusammenfassung .............................................................................................................. 145 9. References .......................................................................................................................... 147 10. Appendix ........................................................................................................................... 165 11. Acknowledgements .......................................................................................................... 167 II Abbreviations 1.Abbreviations A Adenine AA Amino acid ABLV Australian bat lyssavirus ARAV Aravan virus BBLV Bokeloh bat lyssavirus C Cytosine CDC Centers for Disease Control and Prevention CNS Central nervous system DNA Deoxyribonucleic acid dRIT Direct rapid immunohistochemical test DUVV Duvenhage virus EBLV-1 European bat lyssavirus 1 EBLV-2 European bat lyssavirus 2 f.p. Footpad FAO Food and Agriculture Organization of the United Nations FAT Fluorescence antibody test FITC Fluorophore conjugated FLI Friedrich-Loeffler-Institut G Guanine GARC Global Alliance for Rabies Control GBLV Gannoruwa bat lyssavirus Gln Glutamine His Histidine i.m. Intramuscular i.n. Intranasal ICTV International Committee on Taxonomy of Viruses IFN Interferon IGR Intergenic region IKOV Ikoma virus indels Insertions and deletions 1 Abbreviations IRKV Irkut virus kb Kilobase KHUV Khujand virus LBV Lagos bat virus LFD Lateral flow device LLEBV Lleida bat lyssavirus MIT Mouse inoculation test MOKV Mokola virus mRNA Messenger ribonucleic acid N Any base nAchR Nicotinic acetylcholine receptor NASBA Nucleic acid sequence based amplification NCAM Neuronal cell adhesion molecule nm Nanometre nt Nucleotide OIE World Organization for Animal Health p75NTR Low affinity nerve growth factor PEP Post-exposure prophylaxis RABV Rabies virus RNA Ribonucleic acid RNase Ribonuclease RNP Ribonucleoprotein RTCIT Rabies tissue culture infection test RT-LAMP Reverse transcription loop mediated isothermal amplification RT-PCR Reverse transcription polymerase chain reaction s.c. Subcutaneous SHIBV Shimoni bat virus STAT Signal transducer and activator of transcription T Thymine TIS Transcription initiation signal TTS Transcription termination signal UTR Untranslated region 2 Abbreviations W Adenine or Thymine WCBV West Caucasian bat virus WHO World Health Organization Y Cytosine or Thymine 3 Abbreviations 4 Introduction 2.Introduction Rabies, caused by lyssaviruses, is a zoonotic disease responsible for an estimated 59000 human deaths per year and is classified as a neglected zoonotic disease by the World Health Organization (WHO). Since rabies is almost always fatal once clinical signs develop, preventive vaccination of risk groups or prompt application of post exposure prophylaxis is of utmost importance. In most of Europe, rabies has been eliminated
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