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Journal of Clinical Microbiology, July 2000, p. 2756-2759, Vol. 38, No. 7
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.
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
*
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.
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