All Posts Tagged With: "genomic DNA"

Excerpt:

Borrelia burgdorferi sensu lato, carried by Ixodes ticks, is one of the most significant human pathogens, causing Lyme disease. As there is no standardized PCR method for detection and identification of spirochaete 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 and rrs and the non-coding rrs–rrlA region). DNA extracted from 579 Ixodes ricinus ticks was subjected to nested PCR. DNA of the examined spirochaetes was detected in 43 (7.4 %) lysates when the fla gene was used as a molecular marker, in 7 (1.2 %) lysates when using primers complementary to the rrs gene, and in 12 (2.1 %) lysates using primers complementary to the non-coding rrs–rrlA sequence. RFLP analysis based on the fla gene helped identify species from the B. burgdorferi sensu lato complex (B. burgdorferi sensu stricto, Borrelia afzelii, Borrelia garinii, Borrelia valaisiana), detect co-infections, and also identify Borrelia miyamotoi. Therefore, the fla gene is the most sensitive and specific molecular marker for the detection and identification of Borrelia spirochaetes in I. ricinus.

 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.