A comparative analysis of molecular markers for Borrelia spirochetes in Ixodes ricinus
By Linda on Jan 5, 2010 in Infections | comments(0)
Borrelia burgdorferi sensu lato, carried by Ixodes ticks, is one of the
most significant human pathogens responsible for Lyme disease. As there
is no standardized method of polymerase chain reaction (PCR) for
detection and identification of spirochetes’ DNA, we carried out a
comparative analysis using a set of complementary primers for three
regions in the genomic DNA of these bacteria (genes fla, rrs and
non-coding rrs-rrlA region). DNA extracted from 579 Ixodes ricinus ticks
was subjected to nested PCR. DNA of the examined spirochetes was
detected in 43 (7.4 %) lysates when we used fla gene as molecular
marker, in 7 (1.2 %), using primers complementary to the rrs gene, and
in 12 (2.1 %) lysates complementary to the non-coding rrs-rrlA sequence.
Restriction fragment length polymorphism (RFLP) analysis, based on fla
gene, helped identify species from the B. burgdorferi sensu lato (B.
burgdorferi sensu stricto, B. afzelii, B. garinii, B. valaisiana),
detect co-infections, and also identify B. miyamotoi. Therefore the fla
gene is the most sensitive and specific molecular marker for the
detection and identification of Borrelia spirochetes in I. ricinus.
