title: A comparative analysis of molecular markers for the detection and identification of Borrelia spirochaetes in Ixodes ricinus.
abstract: 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.
authors.affiliation: Department of Genetics, University of Szczecin, 71-065 Szczecin, Poland.
authors.forename: Beata
authors.initials: B
authors.lastname: Wodecka
authors.affiliation: Department of Genetics, University of Szczecin, 71-065 Szczecin, Poland.
authors.forename: Agata
authors.initials: A
authors.lastname: Leońska
authors.affiliation: Department of Genetics, University of Szczecin, 71-065 Szczecin, Poland.
authors.forename: Bogumiła
authors.initials: B
authors.lastname: Skotarczak
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D015748:Borrelia burgdorferi Group / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics*; D018884:Ixodes / Q000382:microbiology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
chemical_list: D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial