Tick-borne bacteria in Ixodes ricinus collected in southern Norway evaluated by a commercial kit and established real-time PCR protocols

Ticks are important vectors of human pathogens. The knowledge of disease causing agents harboured by ticks in Norway is limited. The focus of this study was (a) to detect the bacteria of medical importance in ticks collected from the vegetation at locations in the southern part of the country and (b) to evaluate a novel commercially available multiplex PCR based method by comparing results with conventional established real-time PCR protocols. Borrelia burgdorferi sensu lato was confirmed to be the most prevalent pathogen detected (31%) among one hundred individually analysed adult ticks. Borrelia miyamotoi, a spirochete associated with relapsing fever, was detected in one sample. Anaplasma phagocytophilum was found in 4% of the ticks, followed by Rickettsia helvetica which was detected in one sample. Similar pathogen prevalence was also detected in 500 ticks analysed in pools. This is the first report of the spotted fever group Rickettsia in Norway. Francisella tularensis, Bartonella species or Coxiella burnetti was not detected. However, due to the low number of ticks analysed, the possible presence of these pathogens in the region cannot be ruled out. All isolates were screened by at least two different molecular methods for each bacterial target; one commercially available multiplex PCR based tick-borne bacteria flow chip system (Master Diagnostica) and corresponding real-time PCR protocols. The comparison of methods verified that most findings were detected by both methods (71 Borrelia, 15 Anaplasma and 2 Rickettsia), whereas two additional Borrelia and Anaplasma infected samples were detected by the real-time protocols.