Phenotypic and genotypic analysis ofBorrelia spp. isolated fromIxodes ricinus ticks by using electrophoretic chips and real-time polymerase chain reaction

Abstract

The genotype of Borrelia burgdorferi sensu lato was detected in 371 out of 1244 ticks. Borrelia determination was based on partial sequencing of the 16S rRNA gene and real-time polymerase chain reactions for identification and quantitation of ospA and recA genes. Different Borrelia spp. were identified; B. garinii in 40% ticks followed by B. afzelii (36.3%), B. burgdorferi sensu stricto (12.9%), B. valaisiana (3.5%), B. lusitaniae (0.8%), B. bissettii (0.5%) and B. miyamotoi-like (0.5%). Cultivation of 30 borrelia strains in BSK-H medium, among them B. valaisiana, B. bissettii-like and B. miyamotoi-like strains was unique in Czechia. Calibrated microfluidic-based quantification showed differences in the concentration of the nucleic acids and molar mass of the outer surface proteins of different Borrelia spp. with standard sensitivity and specificity and was helpful for their identification. The outer surface protein OspA was absent in B. miyamotoi-like and the OspB protein in B. valaisiana, B. lusitaniae and in three subtypes of B. garinii.

Imbalanced presence of Borrelia burgdorferi s.l. multilocus sequence types in clinical manifestations of Lyme borreliosis

“…complex in Europe (Piesman and Gern, 2004;Rauter and Hartung, 2005). At the opposite pole (with only one MLST profile retrieved from literature) was B. bissettii, a genospecies relatively common in North America (Bissett and Hill, 1987;Oliver et al, 2003), but that has been seldom found in Europe (Fingerle et al, 2008;Hulinska et al, 2007;Strle et al, 1997). The low frequency of B. spielmanii, and B. lusitaniae could be explained by the low relative abundance and patchy habitats of the competent transmission vertebrates for those genospecies -dormice and lizards (Majlathova et al, 2006;Richter and Matuschka, 2006;Richter et al, 2011).…”

16S rRNA gene-based identification of cultured bacterial flora from host-seeking Ixodes ricinus, Dermacentor reticulatus and Haemaphysalis concinna ticks, vectors of vertebrate pathogens

“…Another tick species D.r. is involved in transmision of F. tularensis, C. burnetii, B. canis, and the H.c. tick could transmit Flavivirus of tick-borne encephalitis in European conditions (Hubálek and Rudolf 2007;Hulínská et al 2007). …”

Delineation of a New Species of the Borrelia burgdorferi Sensu Lato Complex, Borrelia americana sp. nov

“…The addition of a 269-nt amplicon of the p66 gene and a 93-nt amplicon of the ospA sequence to the concatenated sequences confirmed the separation of all spirochete species into two groups. (20,35,54,56,57,61). The last decade of the 20th century brought a significant increase in the number of identified Borrelia strains in the United States (1,5,34,37,51,66).…”