A comparison of two DNA extraction approaches in the detection of Borrelia burgdorferi sensu lato from live Ixodes ricinus ticks by PCR and reverse line blotting.
We tested two approaches to extract Borrelia DNA from live Ixodes ricinus ticks before polymerase chain reaction (PCR) and reverse line blotting (RLB): DNA extraction of one half of the tick after incubation in BSK medium and DNA extraction of the other half of the tick directly, using ammonium hydroxide. Among 2079 ticks, 31.2% (n=649) were found to be Borrelia-infected by PCR-RLB test using at least one of the DNA extraction methods. Five hundred four ticks (24.2%) were found infected after incubation in BSK and 481 (23.1%) after direct DNA extraction from the tick. The difference was not significant. However, these prevalences were significantly lower when only one method was applied (23.1% and 24.2%) compared to the prevalence obtained by the use of both methods (31.2%). In 313 infected ticks discordant results were obtained, i.e., one half of the tick was found to be infected whereas the other half was uninfected. Among these ticks, B. garinii and B. burgdorferi sensu stricto (ss) were significantly more frequently identified in the half tick incubated in BSK. No significant differences were observed for B. burgdorferi ss, B. valaisiana, and for undetermined Borrelia species.
Morán-Cadenas
Francisca
F
Institute of Biology, Parasites Eco-Epidemiology Laboratory, University of Neuchâtel, Neuchâtel, Switzerland.
Schneider
Hélène
H
Lommano
Elena
E
Burri
Caroline
C
Moret
Jacqueline
J
Gern
Lise
L
D000818:Animals; D001096:Arachnid Vectors / Q000382:microbiology*; D015139:Blotting, Southern; D015748:Borrelia burgdorferi Group / Q000145:classification / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D018884:Ixodes / Q000382:microbiology*; D016133:Polymerase Chain Reaction
D004269:DNA, Bacterial