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.
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 degrees 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.
Pietilä
J
J
National Public Health Institute, Department in Turku, and Turku Immunology Centre, Turku, Finland.
He
Q
Q
Oksi
J
J
Viljanen
M K
MK
D000818:Animals; D001898:Borrelia / Q000145:classification* / Q000235:genetics; D025065:Borrelia burgdorferi; D015748:Borrelia burgdorferi Group / Q000145:classification* / Q000235:genetics; D005453:Fluorescence; D006801:Humans; D018884:Ixodes / Q000382:microbiology; D008193:Lyme Disease / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011938:Rec A Recombinases / Q000235:genetics*; D013696:Temperature
D011938:Rec A Recombinases