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Journal of Clinical Microbiology, July 2000, p. 2756-2759, Vol. 38, No. 7
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Rapid Differentiation of Borrelia garinii from Borrelia afzelii and Borrelia burgdorferi Sensu Stricto by LightCycler Fluorescence Melting Curve Analysis of a PCR Product of the recA Gene

Johanna Pietilä,1 Qiushui He,1,2,* Jarmo Oksi,3,4 and Matti K. Viljanen1,5

National Public Health Institute, Department in Turku,1 Departments of Pediatrics,2 Medical Microbiology,3 and Internal Medicine,4 University of Turku, and Turku Immunology Centre,5 Turku, Finland

Received 3 January 2000/Returned for modification 25 March 2000/Accepted 20 April 2000

To differentiate the Borrelia burgdorferi sensu lato genospecies, LightCycler real-time PCR was used for the fluorescence (SYBR Green I) melting curve analysis of borrelial recA gene PCR products. The specific melting temperature analyzed is a function of the GC/AT ratio, length, and nucleotide sequence of the amplified product. A total of 32 DNA samples were tested. Of them three were isolated from B. burgdorferi reference strains and 16 were isolated from B. burgdorferi strains cultured from Ixodes ricinus ticks; 13 were directly isolated from nine human biopsy specimens and four I. ricinus tick midguts. The melting temperature of B. garinii was 2°C lower than that of B. burgdorferi sensu stricto and B. afzelii. Melting curve analysis offers a rapid alternative for identification and detection of B. burgdorferi sensu lato genospecies.

* Corresponding author. Mailing address: National Public Health Institute, Department in Turku, Kiinamyllynkatu 13, 20520 Turku, Finland. Phone: 358-2-251 9255. Fax: 358-2-251 9254. E-mail: qiuhe{at}utu.fi.

Journal of Clinical Microbiology, July 2000, p. 2756-2759, Vol. 38, No. 7
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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