Biologia 63/1: 34—37, 2008 Section Cellular and Molecular Biology DOI: 10.2478/s11756-008-0007-1 Comparison of an oligo-chip based assay with PCR method to measure the prevalence of tick-borne pathogenic bacteria in central Slovakia Elena Kocianová 1 , Dušan Blaškovič 2 , Katarína Smetanová 1 , Katarína Schwarzová 1 , Vojtech Boldiš 1 , Zina Košťanová 3 , Denisa M¨ ullerová 2 & Imrich Barák 2 * 1 Institute of Virology, Slovak Academy of Science, Dúbravská cesta 9, SK-84505 Bratislava, Slovakia 2 Institute of Molecular Biology, member of the Centre of Excellence for Molecular Medicine, Slovak Academy of Sciences, SK-84551 Bratislava 45, Slovakia; e-mail: imrich.barak@savba.sk 3 Regional Office of Public Health in Žiar nad Hronom, Sládkovičova 484/9, SK-96524 Žiar nad Hronom, Slovakia Abstract: Ticks are well-known vectors for a wide range of pathogenic microorganisms. We examined the presence of Rickettsia spp., Anaplasma spp., Borrelia spp., Coxiella burnetii and Francisella tularensis in Ixodes ricinus ticks collected in central Slovakia using oligo-chip based assay. Rickettsiae were detected in 5.6% of examined ticks. Borreliae and anaplasmae were identified in 2.1% and 2.8% ticks, respectively. All tested samples were negative for presence of Coxiella burnetii and Francisella tularensis. All these results were compared with those obtained by PCR analysis, and a close correlation between them was found. In addition, rickettsiae of spotted fever group (SFG), Anaplasma phagocytophilum and Borrelia burgdorferi sensu lato were found in ticks using genera or species-specific PCR methods. They are circulating in 10 out of 18 studied localities. Key words: tick-borne pathogenic bacteria; Ixodes ricinus; polymerase chain reaction; DNA-chip. Abbreviations: LB, Lyme borreliosis; SFG, spotted fever group, s.l., sensu lato; s.s., sensu stricto. Introduction Ticks are significant biological vectors of wide spectrum of microorganisms, e.g., viruses, bacteria, and protozoa. Genera Ixodes, Dermacentor and Haemaphysalis are dominant in exophilic ticks in central Europe (Černý 1972; Siuda 1991; Řeháček et al. 1991). Slovakia is a country with abundance of various biotopes for ticks, which range from meadows and forests of Pannonian lowland to Carpathian woods. Diversity of habitats and a wide range of vertebrate tick hosts (birds, small mam- mals, deer and wild boar) create suitable conditions for survival of ticks. Humans may be accidentally bitten by all life stages of Ixodes ricinus that is the vector of various pathogens causing zoonoses. In Europe, I. ricinus is the main vector for Borre- lia burgdorferi sensu lato (s.l.) complex, agent of Lyme borreliosis (LB) (Gern & Humair 2002). High incidence of human LB has been described in Slovakia (pub- lished in the Epidemiological information system of Slo- vakia; http://www.epis.sk/). The LB has been mon- itored since 1987. Average annual morbidity in years 1987–1991 was 4.24 per 100 000 inhabitants. How- ever, increased incidence of LB was observed in last five years, the average annual morbidity in years 2002– 2006 being increased to 13.17 per 100 000 inhabitants. Serological tests of patients with suspected LB con- firmed the infection with these agents in most cases (Kocianová et al. 2005). Routine laboratory diagnosis of tick-borne diseases such as anaplasmosis and rick- ettsioses has not yet been established. Different bacterial pathogens have been detected in ticks collected in Slovakia. Presence of Anaplasma phagocytophilum and B. burgdorferi s.l. in I. ricinus was reported in southwestern and eastern Slovakia (Gern et al. 1999; Špitalská & Kocianová 2002; Derdáková et al. 2003). Rickettsia slovaca has been reported to occur in natural foci in Slovakia since 1968 (Brezina et al. 1969; Mittermayer et al. 1980; Řeháček 1984; Sekeyová et al. 1998). However, other rickettsiae of un- known pathogenicity have been detected in Slovakia and in the neighbouring countries. Unnamed spotted fever group (SFG) Rickettsia was detected in I. ricinus from Austria (Řeháček et al. 1997). Rickettsia IRS3 and IRS4 were isolated from I. ricinus collected in Slovakia (Sekeyová et al. 2000). R. helvetica, R. monacensis, and ‘Candidatus Rickettsia kotlanii’ were recently detected in Hungarian ticks (Sreter-Lancz et al. 2006). * Corresponding author c 2008 Institute of Molecular Biology, Slovak Academy of Sciences Unauthenticated Download Date | 7/26/18 5:09 AM