Genetic Markers for Detecting the DNA of Pathogenic Borrelia

Abstract

The aim of the study was to analyze the genetic markers of Lyme disease pathogens, which can be used to specifically indicate maximum number of their strains and isolates. Materials and methods. The nucleotide sequences of various genes of Borrelia garinii, B. afzelii, B. burgdorferi were downloaded from the NCBI database (National Center for Biological Informatization). The occurrence of the analyzed nucleotide sequences in the genetic code of various organisms was determined in the nBLAST software utility. For the design of primers and probes, the Vector NTI 9.1.0 program (“Invitrogen Corporation”, Carlsbad, USA) was used. DNA was isolated using the MAGNO-sorb kit, version 100-200 (“AmpliSens”, Moscow, Russia), according to the manufacturer’s instructions. Primers and probes were synthesized at “Evrogen” company (Moscow, Russia). For PCR, reagents manufactured by “Synthol” company (Moscow, Russia) were applied.Results and discussion. In order to perform the reliable indication of pathogenic Borrelia, specific loci (genes) of B. garinii, B. afzelii, B. burgdorferi, which were significantly different from the genetic code of other representatives of the genus Borrelia and from the DNA of other organisms, have been determined by molecular-genetic methods. As a result of a preliminary determination of the analytical significance of the studied loci, the following genes and loci were selected for further work: pepX, clpA, ospA, p83/100, ospC and flaB, of which the flaB and ospA genes were selected for practical indication of pathogenic Borrelia DNA. The genetic markers of B. burgdorferi and B. afzelii are displayed during amplification of the flaB gene, while B. garinii and B. afzelii occur when the ospA gene is used as a genetic marker.