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Research article - Peer-reviewed, 2022

A Novel High Discriminatory Protocol for the Detection of Borrelia afzelii, Borrelia burgdorferi Sensu Stricto and Borrelia garinii in Ticks

Chiappa, Giulia; Perini, Matteo; Cafiso, Alessandra; Nodari, Riccardo; Wilhelmsson, Peter; Lindgren, Per-Eric; Omazic, Anna; Ullman, Karin; Moutailler, Sara; Kjellander, Petter; Bazzocchi, Chiara; Grandi, Giulio


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.


Borrelia burgdorferi sensu lato; molecular typing; real time-PCR; Ixodes

Published in

2022, volume: 11, number: 11, article number: 1234
Publisher: MDPI

Authors' information

Chiappa, Giulia
University of Milan
Perini, Matteo
University of Milan
Cafiso, Alessandra
University of Milan
Nodari, Riccardo
University of Milan
Wilhelmsson, Peter
Linkoping University
Lindgren, Per-Eric
Linkoping University
Omazic, Anna
National Veterinary Institute SVA
Ullman, Karin
National Veterinary Institute SVA
Moutailler, Sara
Swedish University of Agricultural Sciences, Department of Ecology
Bazzocchi, Chiara
University of Milan
National Veterinary Institute SVA

UKÄ Subject classification

Biological Systematics

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