Identification of Three Clinically Relevant Borrelia burgdorferi Sensu Lato Genospecies by PCR-Restriction Fragment Length Polymorphism Analysis of 16S-23S Ribosomal DNA Spacer Amplicons

Abstract

We report the results of a study of the prevalences of three clinically relevant Borrelia burgdorferi sensu lato genospecies (Borrelia burgdorferi sensu stricto, Borrelia afzelii, and Borrelia garinii) in 1,040 questing Ixodes ticks from all regions of Latvia, where Lyme borreliosis is endemic. The prevalences of Borrelia in Ixodes ricinus and Ixodes persulcatus were 22.6 and 27.9%, respectively. Molecular typing of B. burgdorferi from infected ticks was performed by restriction fragment length polymorphism (RFLP) analysis of PCR-amplified fragments of the 16S-23S (rrs-rrlA) rRNA intergenic spacer by using species-specific primers and subsequent sequencing. The dominant Borrelia species in both Ixodes species was B. afzelii. In addition, different restriction patterns of B. garinii and B. afzelii were also identified. This study demonstrates that the 16S-23S rRNA PCR-RFLP typing method is simple, sensitive, and fast and that it allows one to differentiate among B. burgdorferi species and subspecies with various degrees of pathogenic potential directly in ticks. These features are important in monitoring Lyme disease.

Capillary electrophoresis and the clinical laboratory

“…For bacteria, the conserved 16S rDNA sequence is often targeted for sequencing [220]. This application has been used to verify the identity of bacteria, and strains or biotypes and has been helpful in cases where rare isolates have been identified [221][222][223][224]. By aligning sequence data, the Mycobacterium family tree has been restructured with the final result being the regrouping of Mycobacterium species and Borreli species [225,226].…”

Genospecies of Borrelia burgdorferi sensu lato in Ixodes ricinus ticks from the Autonomous Province of Trento, Italy

“…Up to now, 10 genospecies of B. burgdorferi s.l. have been isolated from ticks in Europe, only five of which cause clinical manifestations of LB in humans: B. burgdorferi s.s., B. garinii, B. afzelii, B. valaisiana and B. lusitaniae (see Wang et al, 1999 for review;Derda´kova´et al, 2003 and references therein; Ranka et al, 2004). Two additional genospecies (B. bissettii and an as yet un-named genospecies) have been found in patients with symptoms of LB (Gern and Humair, 2002).…”

Detection of Yersinia enterocolitica in milk powders by cross-priming amplification combined with immunoblotting analysis

“…This genomic region shows a significant variation among different species of bacteria, both in sequence and length (Hurtle et al, 2003). Bacteria can harbor multiple alleles of ribosomal operon in their genome, resulting in a significant sequence variation in the spacer region among different species and even strains (Kabadjova et al, 2002;Rachman et al, 2004;Ranka et al, 2004). Because a few Yersinia species (e.g., Yersinia intermedia and Yersinia kristensenii) are similar to Y. enterocolitica in their 16S rDNA sequence, it is critical to develop a specific, rapid and sensitive method for the accurate detection of Y. enterocolitica.…”