Phylogenetic Analysis of Borrelia burgdorferi Sensu Stricto by Arbitrarily Primed PCR and Pulsed-Field Gel Electrophoresis

Foretz, Marc; Postić, D.; Baranton, Guy

doi:10.1099/00207713-47-1-11

Cited by 35 publications

(36 citation statements)

References 24 publications

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“…We observed greater genetic diversity among the 15 North American samples (mean genetic distance ϭ 0.46) than among 12 European samples (mean genetic distance ϭ 0.41). Perhaps because of a relatively small sample size, this trend is not statistically significant (t ϭ 0.009) but it is consistent with previous evolutionary models postulating a founder effect as North American B. burgdorferi sensu stricto moved to the Old World (15,23). However, diversity within B. burgdorferi sensu stricto could likewise be affected by lateral transfer of genetic material from other species.…”

Section: Resultssupporting

confidence: 71%

“…Numerous molecular techniques have recently been used to characterize Borrelia species, including 16S rRNA gene sequence analysis, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Western blot analysis, pulsed-field gel electrophoresis, plasmid fingerprinting, randomly amplified polymor-phic DNA analysis, restriction fragment length polymorphism analysis, fatty acid profile analysis, and serotyping (4,8,15,33). For a more thorough review of the molecular typing methods used for Borrelia characterization, see reference 36.…”

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confidence: 99%

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Strain Typing of Borrelia burgdorferi , Borrelia afzelii , and Borrelia garinii by Using Multiple-Locus Variable-Number Tandem Repeat Analysis

et al. 2002

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Human Lyme borreliosis (LB) is the most prevalent arthropod-borne infection in temperate climate zones around the world and is caused by Borrelia spirochetes. We have identified 10 variable-number tandem repeat (VNTR) loci present within the genome of Borrelia burgdorferi and subsequently developed a multiple-locus VNTR analysis (MLVA) typing system for this disease agent. We report here the successful application of MLVA for strain discrimination among a group of 41 globally diverse Borrelia isolates including B. burgdorferi, B. afzelii, and B. garinii. PCR assays displayed diversity at these loci, with total allele numbers ranging from two to nine and Nei's diversity (

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Section: Resultssupporting

confidence: 71%

mentioning

confidence: 99%

Strain Typing of Borrelia burgdorferi , Borrelia afzelii , and Borrelia garinii by Using Multiple-Locus Variable-Number Tandem Repeat Analysis

et al. 2002

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“…Numerous studies have demonstrated substantial genotypic variability among B. burgdorferi isolates (23,39,40,77). Thus, the extent to which these two isolates are representative of all virulent and attenuated strains is unclear, although increased expression of BBA74 was observed in the two attenuated strains studied thus far (B356 and B331).…”

Section: Discussionmentioning

confidence: 99%

Comparative Transcriptional Profiling of Borrelia burgdorferi Clinical Isolates Differing in Capacities for Hematogenous Dissemination

et al. 2005

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Borrelia burgdorferi, the etiologic agent of Lyme disease, is genetically heterogeneous. Previous studies have shown a significant association between the frequency of hematogenous dissemination in Lyme disease patients and the genotype of the infecting B. burgdorferi strain. Comparative transcriptional profiling of two representative clinical isolates with distinct genotypes (BL206 and B356) was undertaken. A total of 78 open reading frames (ORFs) had expression levels that differed significantly between the two isolates. A number of genes with potential involvement in nutrient uptake (BB0603, BBA74, BB0329, BB0330, and BBB29) have significantly higher expression levels in isolate B356. Moreover, nearly 25% of the differentially expressed genes are predicted to be localized on the cell surface, implying that these two isolates have cell surface properties that differ considerably. One of these genes, BBA74, encodes a protein of 257 amino acid residues that has been shown to possess porin activity. BBA74 transcript level was >20-fold higher in B356 than in BL206, and strain B356 contained three-to fivefold more BBA74 protein. BBA74 was disrupted by the insertion of a kanamycin resistance cassette into the coding region. The growth rates of both wild-type and mutant strains were essentially identical, and cultures reached the same final cell densities. However, the mutant strains consistently showed prolonged lags of 2 to 5 days prior to the induction of log-phase growth compared to wild-type strains. It is tempting to speculate that the absence of BBA74 interferes with the enhanced nutrient uptake that may be required for the entry of cells into log-phase growth. These studies demonstrate the value of comparative transcriptional profiling for identifying differences in the transcriptomes of B. burgdorferi clinical isolates that may provide clues to pathogenesis. The 78 ORFs identified here are a good starting point for the investigation of factors involved in the hematogenous dissemination of B. burgdorferi.

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“…Previously described procedures were used for the preparation of high-molecular-weight genomic DNAs and for PFGE (4,9). Rare-cutting endonucleases SacII, BssHII, and MluI were used to differentiate isolates.…”

Section: Methodsmentioning

confidence: 99%

Characterization of Borrelia lusitaniae Isolates Collected in Tunisia and Morocco

et al. 2005

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Borrelia lusitaniae is a species within the complex Borrelia burgdorferi sensu lato and is infrequently isolated in Europe. In contrast, this species is by far the most predominant in North Africa and in Portugal. In this study, we analyzed the genetic diversity, at several loci, of a large population of isolates from free-living Ixodes ricinus ticks collected in Tunisia and Morocco. We found a moderate diversity of the whole genome by using pulsed-field gel electrophoresis as well as in the ospA gene sequences, compared to a high level of strain homogeneity in the small noncoding ribosomal spacer. In contrast, a high diversity of this locus has been previously reported for Portuguese isolates. We hypothesize that B. lusitaniae strains isolated in North Africa constitute a clone of Portuguese origin.Within the complex Borrelia burgdorferi sensu lato, three species, B. burgdorferi sensu stricto, Borrelia garinii, and Borrelia afzelii, are known to cause a broad spectrum of human manifestations that are described as Lyme borreliosis. The genetic diversity of pathogenic species has been largely investigated both at the species level and at the strain level within each species (41). Otherwise, although its pathogenicity for humans is only suspected (31, 33), Borrelia valaisiana represents a widely distributed species in ticks throughout Europe and Asia and therefore has been extensively studied (12,22,32,38,40).In contrast to these species, little data are available concerning Borrelia lusitaniae. Only sporadic isolates have been characterized from ticks collected in the Czech Republic, Moldavia, Ukraine, and Belarus (20), in Slovakia (11), in Spain (3), in Poland (25), in France (30), in Switzerland (15-17), or in Turkey (13), and these isolates did not show any significant diversity. However, a study conducted in Portugal revealed a large diversity among B. lusitaniae strains isolated from a local tick population (6). Moreover, it is noteworthy that, among the three first Portuguese isolates identified as B. lusitaniae, two different types were demonstrated (20,26). Hence, these studies suggested a higher diversity among B. lusitaniae around the Mediterranean basin. However, all these studies only focused on the small ribosomal rrf-rrl spacer and, therefore, the global diversity of this species remained unknown.Recently, we reported that B. lusitaniae was, by far, the most prevalent species, ranging from 96.6 to 100% of the B. burgdorferi sensu lato species identified in Tunisia and Morocco (35,44,46). The report of some Lyme borreliosis cases in North Africa (1, 27) and the recent isolation in Portugal of B.lusitaniae from the skin of a human patient with chronic lesions (5) prompted us to characterize this species as a potential human pathogen. Therefore, we investigated the molecular characterization and the genetic diversity of B. lusitaniae isolated from free-living Ixodes ricinus ticks in Tunisia and Morocco. For these purposes we studied two single loci (ospC and ospA) and the noncoding intergenic spacer (r...

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Phylogenetic Analysis of Borrelia burgdorferi Sensu Stricto by Arbitrarily Primed PCR and Pulsed-Field Gel Electrophoresis

Cited by 35 publications

References 24 publications

Strain Typing of Borrelia burgdorferi , Borrelia afzelii , and Borrelia garinii by Using Multiple-Locus Variable-Number Tandem Repeat Analysis

Strain Typing of Borrelia burgdorferi , Borrelia afzelii , and Borrelia garinii by Using Multiple-Locus Variable-Number Tandem Repeat Analysis

Comparative Transcriptional Profiling of Borrelia burgdorferi Clinical Isolates Differing in Capacities for Hematogenous Dissemination

Characterization of Borrelia lusitaniae Isolates Collected in Tunisia and Morocco

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