Biomedical Research and Study Centre University of Latvia Riga, Latvia Molecular biology of Borrelia burgdorferi sensu lato in Latvia Renate Ranka Academic dissertation 2004 /-- Supervisor: ~ Professor, Dr.BioI., MD Viesturs Baumanis Opponents: Professor Sven Bergstrom, Urnea University, Sweden 1: Professor, Dr.Hab.Biol. Pauls Pumpens, University of Latvia Professor, Dr.Med. Juta Kroica, Riga Stradin's University, Latvia. Author's address: Biomedical Research and Study Centre, University of Latvia, Laboratory of Molecular Microbiology Ratsupites str 1, Riga, LATVIA, LV-1067 Fax: 371 7442407; Tel: 371 7808218 e-mail:
[email protected] TABLE OF CONTENTS 1. LIST OF ABBREVIA nONS AND TERMS 3 2. INTR 0DUerr ON 4 3. PAPERS IN THIS THESIS 6 4. REVIEW OF THE LITERA TURE 7 4.1. Main characteristics of Borrelia burgdorferi spirochetes 7 4.2. Molecular biology of Borrelia burgdorferi sensu lato 9 4.3. Natural cycle and vectors of Borrelia burgdorferi 11 4.4. Lyme disease \-l 4.5. Immunologic testing in diagnosis of Lyme disease 16 4.6. Borrelia burgdorferi proteins and their immunogenetic properties 17 5. AIMS OF 'nus THESIS 20 6. METHODS 21 6.1. Collection of ticks 21 6.2. Microscopic detection of Borrelia in questing ticks 21 6.3. Cultivation of Borrelia burgdorferi 21 6.4. Reference DNA 21 6.5. DNA extraction 22 6.6. B. burgdorferi detection by PCR amplification 22 6.7. Molecular typing of B. burgdorferi by 16S-23S ribosomal DNA spacer PCR- RFLP 23 6.8. Molecular typing of B. burgdorferi by species-specific PCR targeted 16S rRNA gene "" 23 6.9.