Borrelia persica: In vitro cultivation and characterization via conventional PCR and multilocus sequence analysis of two strains isolated from a cat and ticks from Israel

Abstract

Borrelia persica, one of the pathogenic agents of tick-borne relapsing fever, is transmitted by the soft tick Ornithodoros tholozani. It causes infections in humans as well as in animals. In this study, we developed a medium, termed Pettenkofer/LMU Bp, for reliable in vitro cultivation. Cell densities up to 5.2×10(7) viable cells/ml were achieved over at least 40 passages. The cultivable B. persica strain isolated from a cat was further analyzed by amplification of the flaB gene using conventional PCR. In addition, seven housekeeping genes (clpA, clpX, pepX, pyrG, recG, rplB and uvrA) of this B. persica strain and a second strain isolated out of pooled ticks from Israel were amplified and the phylogenetic relationships among Borrelia species were analyzed. The results of the conventional PCR and the multilocus sequence analysis confirmed our isolates as B. persica.

Keywords: Borrelia persica; In vitro cultivation; MLSA; MLST; PCR; Relapsing fever spirochete.