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Journal of Clinical Microbiology, April 2004, p. 1444-1449, Vol. 42, No. 4
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.4.1444-1449.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

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

Renate Ranka,1* Antra Bormane,2 Kristine Salmina,1 and Viesturs Baumanis1

Biomedical Research and Study Centre, University of Latvia, LV-1067,1 State Public Health Agency, LV-1012, Riga, Latvia2

Received 19 March 2003/ Returned for modification 19 August 2003/ Accepted 7 January 2004

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.

* Corresponding author. Mailing address: Biomedical Research and Study Centre, Ratsupites St. 1, Riga, Latvia LV-1067. Phone: 371 7808218. Fax: 371 7442407. E-mail: renate_r{at}biomed.lu.lv.

Journal of Clinical Microbiology, April 2004, p. 1444-1449, Vol. 42, No. 4
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.4.1444-1449.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.





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