A comparative analysis of molecular markers for the detection and identificatio

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J Med Microbiol.. [Epub ahead of print]

 

A comparative analysis of molecular markers for the detection and

identification of Borrelia spirochetes in Ixodes ricinus.

 

Wodecka B, Leonska A, Skotarczak B.

 

University of Szczecin.

 

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.

 

PMID: 20007765 [PubMed - as supplied by publisher]