A novel duplex real-time PCR permits simultaneous detection and differentiation of Borrelia miyamotoi and Borrelia burgdorferi sensu lato

Venczel, R.; Knoke, Lisa R.; Pavlovic, Melanie; Dzaferovic, Eldina; Vaculová, Tatiana; Silaghi, Cornelia; Overzier, Evelyn; Konrad, Regina; Kolencik, Stanislav; Derdáková, Markéta; Sing, Andreas; Schaub, Günter A.; Margos, Gabriele; Fingerle, Volker

doi:10.1007/s15010-015-0820-8

Cited by 39 publications

(23 citation statements)

References 35 publications

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“…known to harbor the spirochete and to bite humans. Others have developed PCR-based assays to detect B. miyamotoi in ticks (e.g., Dibernardo et al, 2014; Houck et al, 2011; Mukhacheva and Kovalev, 2014; Quarsten et al, 2015; Reiter et al, 2015; Tsao et al, 2004; Ullmann et al, 2005; Vayssier-Taussat et al, 2013; Venczel et al, 2016). The approach described here has the advantage of integrating two sensitive, species-specific targets designed to detect genetically diverse North American strains of B. miyamotoi with a tick DNA target that can be used to verify the integrity of DNA derived from I. scapularis or I. pacificus .…”

Section: Resultsmentioning

confidence: 99%

Paired real-time PCR assays for detection of Borrelia miyamotoi in North American Ixodes scapularis and Ixodes pacificus (Acari: Ixodidae)

Graham

Pilgard

Maes

et al. 2016

Ticks and Tick-borne Diseases

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Borrelia miyamotoi is an emerging, tick-borne human pathogen. In North America, it is primarily associated with Ixodes scapularis and Ixodes pacificus, two species known to bite humans. Here we describe the development and evaluation of a pair of real-time TaqMan PCR assays designed to detect B. miyamotoi in North American ticks. We sought to achieve sensitivity to B. miyamotoi strains associated with ticks throughout North America, the full genetic diversity of which is unknown, by targeting sequences that are largely conserved between B. miyamotoi strains from the eastern United States and genetically distinct B. miyamotoi strains from Japan. The two assays target different loci on the B. miyamotoi chromosome and can be run side by side under identical cycling conditions. One of the assays also includes a tick DNA target that can be used to verify the integrity of tick-derived samples. Using both recombinant plasmid controls and genomic DNA from North American and Japanese strains, we determined that both assays reliably detect as few as 5 copies of the B. miyamotoi genome. We verified that neither detects B. burgdorferi, B. lonestari or B. turicatae. This sensitive and specific pair of assays successfully detected B. miyamotoi in naturally-infected, colony-reared nymphs and in field-collected I. scapularis and I. pacificus from the Northeast and the Pacific Northwest respectively. These assays will be useful in screening field-collected Ixodes spp. from varied regions of North America to assess the risk of human exposure to this emerging pathogen.

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Section: Resultsmentioning

confidence: 99%

Paired real-time PCR assays for detection of Borrelia miyamotoi in North American Ixodes scapularis and Ixodes pacificus (Acari: Ixodidae)

Graham

Pilgard

Maes

et al. 2016

Ticks and Tick-borne Diseases

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“…In all cases, a preliminary diagnosis was established by clinicians based on clinical symptoms and visible spirochetes in Giemsa stained blood films The German National Reference Centre (NRC) for Borrelia in Oberschleißheim was contacted directly by clinicians for confirmation. Microbiological confirmation of LBRF from EDTA blood included: Dark field microscopy (Leitz Dialux 20 microscope, Leitz, Germany, objective 40, occular 10) and PCR targeting 16S rRNA, flab, and glpQ followed by sequencing [1][2][3].…”

Section: Case Investigationmentioning

confidence: 99%

Louse-borne relapsing fever (Borrelia recurrentis) diagnosed in 15 refugees from northeast Africa: epidemiology and preventive control measures, Bavaria, Germany, July to October 2015

Hoch¹,

Wieser

Löscher³

et al. 2015

Eurosurveillance

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We report 15 imported louse-borne relapsing fever (LBRF) cases in refugees in Bavaria, Germany. One patient died. Epidemiological findings confirmed that all were young males from the Horn of Africa (12 from Somalia), who had similar migration routes converging in Sudan continuing through Libya and Italy. The majority likely acquired their infection during migration. Healthcare workers should be aware of LBRF in refugees passing through north Africa to ensure correct treatment and preventive measures.

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“…The prevalence of B. miyamotoi in ticks was found to be usually lower than that of B. burgdorferi s.l. although prevalences of~15% have been reported in some regions [3,7,10,16,17,21,22]. Rodents have been implicated as reservoir hosts for B. miyamotoi [23,24], but transovarial transmission is also known to occur [25,26] and may contribute to the persistence of this Borrelia in nature.…”

Section: Introductionmentioning

confidence: 99%

Whole genome sequencing of Borrelia miyamotoi isolate Izh-4: reference for a complex bacterial genome

Kuleshov

Margos²,

Fingerle³

et al. 2020

BMC Genomics

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Background: The genus Borrelia comprises spirochaetal bacteria maintained in natural transmission cycles by tick vectors and vertebrate reservoir hosts. The main groups are represented by a species complex including the causative agents of Lyme borreliosis and relapsing fever group Borrelia. Borrelia miyamotoi belongs to the relapsing fever group of spirochetes and forms distinct populations in North America, Asia, and Europe. As all Borrelia species B. miyamotoi possess an unusual and complex genome consisting of a linear chromosome and a number of linear and circular plasmids. The species is considered an emerging human pathogen and an increasing number of human cases are being described in the Northern hemisphere. The aim of this study was to produce a high quality reference genome that will facilitate future studies into genetic differences between different populations and the genome plasticity of B. miyamotoi. Results: We used multiple available sequencing methods, including Pacific Bioscience single-molecule real-time technology (SMRT) and Oxford Nanopore technology (ONT) supplemented with highly accurate Illumina sequences, to explore the suitability for whole genome assembly of the Russian B. miyamotoi isolate, Izh-4. Plasmids were typed according to their potential plasmid partitioning genes (PF32, 49, 50, 57/62). Comparing and combining results of both long-read (SMRT and ONT) and short-read methods (Illumina), we determined that the genome of the isolate Izh-4 consisted of one linear chromosome, 12 linear and two circular plasmids. Whilst the majority of plasmids had corresponding contigs in the Asian B. miyamotoi isolate FR64b, there were only four that matched plasmids of the North American isolate CT13-2396, indicating differences between B. miyamotoi populations. Several plasmids, e.g. lp41, lp29, lp23, and lp24, were found to carry variable major proteins. Amongst those were variable large proteins (Vlp) subtype Vlp-α, Vlp-γ, Vlp-δ and also Vlp-β. Phylogenetic analysis of common plasmids types showed the uniqueness in Russian/Asian isolates of B. miyamotoi compared to other isolates. Conclusions: We here describe the genome of a Russian B. miyamotoi clinical isolate, providing a solid basis for future comparative genomics of B. miyamotoi isolates. This will be a great impetus for further basic, molecular and epidemiological research on this emerging tick-borne pathogen.

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A novel duplex real-time PCR permits simultaneous detection and differentiation of Borrelia miyamotoi and Borrelia burgdorferi sensu lato

Abstract: The duplex real-time PCR developed here represents a method that permits simultaneous detection and differentiation of B. burgdorferi s.l. and B. miyamotoi in environmental and potentially clinical samples.

Cited by 39 publications

References 35 publications

Paired real-time PCR assays for detection of Borrelia miyamotoi in North American Ixodes scapularis and Ixodes pacificus (Acari: Ixodidae)

Paired real-time PCR assays for detection of Borrelia miyamotoi in North American Ixodes scapularis and Ixodes pacificus (Acari: Ixodidae)

Louse-borne relapsing fever (Borrelia recurrentis) diagnosed in 15 refugees from northeast Africa: epidemiology and preventive control measures, Bavaria, Germany, July to October 2015

Whole genome sequencing of Borrelia miyamotoi isolate Izh-4: reference for a complex bacterial genome

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