First Isolation of
            <i>Borrelia lusitaniae</i>
            from a Human Patient

Collares-Pereira, Margarida; Couceiro, Sofia; Franca, Isabel da; Kurtenbach, Klaus; Schäfer, Stefanie; Vitorino, Liliana; Gonçalves, Luis Roberto de Camargo; Baptista, Susana; Vieira, Maria Luísa; Cunha, Celso

doi:10.1128/jcm.42.3.1316-1318.2004

Cited by 180 publications

(110 citation statements)

References 11 publications

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“…That group comprises the species B. burgdorferi sensu stricto, B. afzelii, and B. garinii, which are usually transmitted by the vector Ixodes ricinus. Furthermore, there have been reports of B. valaisiana, B. lusitaniae, and B. spielmanii being detected in samples of human skin and cerebrospinal fluid (5,7,30), which suggests that those three species can also give rise to LB. It is often hard to distinguish the clinical symptoms of LB from those of other diseases (10), and hence, it can be difficult to establish a correct diagnosis, especially if the patient is unable to recall having a tick bite.…”

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confidence: 99%

Prevalence and Diversity ofBorreliaSpecies in Ticks That Have Bitten Humans in Sweden

et al. 2010

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Members of the genus Borrelia are among the most common infectious agents causing tick-borne disease in humans worldwide. Here, we developed a Light Upon eXtension (LUX) real-time PCR assay that can detect and quantify Borrelia species in ticks that have fed on humans, and we applied the assay to 399 such ticks. Borrelia PCR-positive ticks were identified to species level by sequencing the products of conventional PCR performed using Borrelia group-specific primers. There was a 19% prevalence of Borrelia spp. in the detached ticks, and the number of spirochetes per Borrelia PCR-positive tick ranged from 2.0 ؋ 10 2 to 4.9 ؋ 10 5 , with a median of 7.8 ؋ 10 3 spirochetes. Adult ticks had a significantly larger number of spirochetes, with a median of 8.4 ؋ 10 4 compared to the median of nymphs of 4.4 ؋ 10 4 . Adult ticks also exhibited a higher prevalence of Borrelia (33%) than nymphs (14%). Among the identified species, Borrelia afzelii was found to predominate (61%) and was followed by B. garinii (23%), B. valaisiana (13%), B. burgdorferi sensu stricto (1%), B. lusitaniae (1%), and B. miyamotoi-like (1%). Also, 3% of the ticks were coinfected with multiple strains of B. afzelii. Notably, this is the first report of B. lusitaniae being detected in ticks in Sweden. Our LUX real-time PCR assay proved to be more sensitive than a corresponding TaqMan assay. In conclusion, the novel LUX real-time PCR method is a rapid and sensitive tool for detection and quantification of Borrelia spp. in ticks.Lyme borreliosis (LB) is the most common tick-borne disease in humans in Europe (26), and it is caused by spirochetes belonging to the Borrelia burgdorferi sensu lato complex. That group comprises the species B. burgdorferi sensu stricto, B. afzelii, and B. garinii, which are usually transmitted by the vector Ixodes ricinus. Furthermore, there have been reports of B. valaisiana, B. lusitaniae, and B. spielmanii being detected in samples of human skin and cerebrospinal fluid (5, 7, 30), which suggests that those three species can also give rise to LB. It is often hard to distinguish the clinical symptoms of LB from those of other diseases (10), and hence, it can be difficult to establish a correct diagnosis, especially if the patient is unable to recall having a tick bite.Today, diagnosis is based mainly on serological tests, although some PCR-based approaches, such as the TaqMan real-time PCR assay (3, 12), have been developed to detect Borrelia species in clinical samples. Even if real-time PCR is not yet considered to be a routine method in clinical practice, it can nonetheless provide valuable information about Borrelia infections, with regard to species type and the number of spirochetes present. Additional major advantages of PCR in this context are its simplicity, sensitivity, robustness, and speed.Other assays besides the TaqMan assay include a method based on SYBR green dye chemistry (37) and another using Light Upon eXtension (LUX) (Invitrogen Corporation). Compared to the SYBR green real-time PCR assay, the LUX assay o...

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confidence: 99%

Prevalence and Diversity ofBorreliaSpecies in Ticks That Have Bitten Humans in Sweden

et al. 2010

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“…Moreover, a recent study indicated that the inoculation of B. lusitaniae into susceptible mice induced pathological features (45). Finally, B. lusitaniae was recently isolated, for the first time, from a chronic skin lesion in a Portuguese patient (5). If this species is actually pathogenic for humans, the risk of Lyme borreliosis must be underlined in North Africa.…”

Section: Discussionmentioning

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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“…Furthermore, recent studies have identified Borrelia lusitaniae as a potential agent of Lyme disease. The first human isolate of B. lusitaniae was identified in a 46-year-old woman from the Lisbon area in Portugal (Collares-Pereira et al 2004). Such genospecies are the most prevalent in the Mediterranean area (Collares-Pereira et al 2004;Sarih et al 2003;Younsi et al 2005) and recent studies indicate lizards as the reservoir hosts (Amore et al 2007;Majlathova et al 2006;Richter and Matuschka 2006;Tomassone et al 2005;Younsi et al 2005).…”

Section: Introductionmentioning

confidence: 99%

Borrelia lusitaniae OspA Gene Heterogeneity in Mediterranean Basin Area

Grego

Bertolotti

Peletto³

et al. 2007

J Mol Evol

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In this study, Borrelia lusitaniae DNA extracted from ticks and lizards was used to amplify the outer surface protein A (OspA) gene in order to increase knowledge about sequence variability in the Mediterranean basin area, to better understand how Borrelia lusitaniae has evolved and how its distribution has expanded. Phylogenetic trees including Italian and reference sequences showed a clear separation of B. lusitaniae OspA strains in two different major clades. North African isolates form a clade with Portuguese POTIB strains, whereas Italian samples are grouped with German strains and a human Portuguese strain. This subdivision was supported by very high posterior probability values in the trees, by both analysis of molecular variance and selective pressure. These results, based on phylogenetic information contained in the OspA gene sequences, show the presence of two different B. lusitaniae strains circulating in the Mediterranean basin area, suggesting two different evolution paths.

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First Isolation of Borrelia lusitaniae from a Human Patient

Cited by 180 publications

References 11 publications

Prevalence and Diversity ofBorreliaSpecies in Ticks That Have Bitten Humans in Sweden

Prevalence and Diversity ofBorreliaSpecies in Ticks That Have Bitten Humans in Sweden

Characterization of Borrelia lusitaniae Isolates Collected in Tunisia and Morocco

Borrelia lusitaniae OspA Gene Heterogeneity in Mediterranean Basin Area

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