A Novel High Discriminatory Protocol for the Detection of <i>Borrelia afzelii</i>, <i>Borrelia burgdorferi</i> Sensu Stricto and <i>Borrelia garinii</i> in Ticks
Giulia Chiappa,
Matteo Perini,
Alessandra Cafiso,
Riccardo Nodari,
Peter Wilhelmsson,
Per-Eric Lindgren,
Anna Omazic,
Karin Ullman,
Sara Moutailler,
Petter Kjellander,
Chiara Bazzocchi,
Giulio Grandi
Affiliations
Giulia Chiappa
Department of Veterinary Medicine and Animal Science, University of Milan, 26900 Lodi, Italy
Matteo Perini
Romeo and Enrica Invernizzi Pediatric Research Center, Department of Biomedical and Clinical Sciences L. Sacco, University of Milan, 20157 Milan, Italy
Alessandra Cafiso
Department of Veterinary Medicine and Animal Science, University of Milan, 26900 Lodi, Italy
Riccardo Nodari
Romeo and Enrica Invernizzi Pediatric Research Center, Department of Biomedical and Clinical Sciences L. Sacco, University of Milan, 20157 Milan, Italy
Peter Wilhelmsson
Department of Biomedical and Clinical Sciences, Division of Inflammation and Infection, Linköping University, 581 83 Linköping, Sweden
Per-Eric Lindgren
Department of Biomedical and Clinical Sciences, Division of Inflammation and Infection, Linköping University, 581 83 Linköping, Sweden
Anna Omazic
Department of Chemistry, Environment and Feed Hygiene, National Veterinary Institute, 751 89 Uppsala, Sweden
Karin Ullman
Department of Microbiology, National Veterinary Institute, 751 89 Uppsala, Sweden
Sara Moutailler
ANSES, Laboratoire de Santé Animale, Ecole Nationale Vétérinaire d’Alfort, INRAE, UMR BIPAR, F-94700 Maisons-Alfort, France
Petter Kjellander
Grimsö Wildlife Research Station, Department of Ecology, Swedish University of Agricultural Sciences, 730 91 Riddarhyttan, Sweden
Chiara Bazzocchi
Department of Veterinary Medicine and Animal Science, University of Milan, 26900 Lodi, Italy
Giulio Grandi
Department of Microbiology, National Veterinary Institute, 751 89 Uppsala, Sweden
Bacteria of the Borrelia burgdorferi sensu lato complex are the causative agents of Lyme borreliosis (LB). Even if the conventional diagnosis of LB does not rely on the species itself, an accurate species identification within the complex will provide a deepened epidemiological scenario, a better diagnosis leading to a more targeted therapeutic approach, as well as promote the general public’s awareness. A comparative genomics approach based on the 210 Borrelia spp. genomes available in 2019 were used to set up three species-specific PCR protocols, able to detect and provide species typing of Borrelia afzelii, Borrelia burgdorferi sensu stricto (s.s.) and Borrelia garinii, the three most common and important human pathogenic Lyme Borrelia species in Europe. The species-specificity of these protocols was confirmed on previously identified B. afzelii, B. burgdorferi s.s. and B. garinii specimens detected in Ixodes ricinus samples. In addition, the protocols were validated on 120 DNA samples from ticks collected in Sweden, showing 88% accuracy, 100% precision, 72% sensitivity and 100% specificity. The proposed approach represents an innovative tool in epidemiological studies focused on B. burgdorferi s.l. occurrence in ticks, and future studies could suggest its helpfulness in routine diagnostic tests for health care.
