PubMLST.org – The new home for the Borrelia MLSA database

Margos, Gabriele; Binder, Katrin; Dzaferovic, Eldina; Hizo-Teufel, Cecilia; Sing, Andreas; Wildner, Manfred; Fingerle, Volker; Jolley, Keith A.

doi:10.1016/j.ttbdis.2015.06.007

Cited by 26 publications

(18 citation statements)

References 11 publications

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“…The BigDye Xterminator Kit (ThermoFisher) was used to remove unincorporated dyes before analyzing the samples on an ABI 3130XL genetic analyzer. We used Lasergene 12 software (DNASTAR, Madison) to construct a consensus sequence for each amplicon based on at least double coverage of every nt and queried the GenBank and pubMLST (http://pubmlst.org/borrelia/; Margos et al 2015) (accessed July 2016) databases for similar sequences. We verified the species identification of all samples that tested positive for B. miyamotoi by using a similar approach, described in Graham et al (2016), to amplify, sequence, and analyze a segment of the clpA gene.…”

Section: Methodsmentioning

confidence: 99%

Prevalence and Diversity of Tick-Borne Pathogens in NymphalIxodes scapularis(Acari: Ixodidae) in Eastern National Parks

Johnson¹,

Graham²,

Boegler³

et al. 2016

J Med Entomol

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Tick-borne pathogens transmitted by Ixodes scapularis Say (Acari: Ixodidae), also known as the deer tick or blacklegged tick, are increasing in incidence and geographic distribution in the United States. We examined the risk of tick-borne disease exposure in 9 national parks across six Northeastern and Mid-Atlantic States and the District of Columbia in 2014 and 2015. To assess the recreational risk to park visitors, we sampled for ticks along frequently used trails and calculated the density of I. scapularis nymphs (DON) and the density of infected nymphs (DIN). We determined the nymphal infection prevalence of I. scapularis with a suite of tick-borne pathogens including Borrelia burgdorferi, Borrelia miyamotoi, Anaplasma phagocytophilum, and Babesia microti. Ixodes scapularis nymphs were found in all national park units; DON ranged from 0.40 to 13.73 nymphs per 100 m2. Borrelia burgdorferi, the causative agent of Lyme disease, was found at all sites where I. scapularis was documented; DIN with B. burgdorferi ranged from 0.06 to 5.71 nymphs per 100 m2. Borrelia miyamotoi and A. phagocytophilum were documented at 60% and 70% of the parks, respectively, while Ba. microti occurred at just 20% of the parks. Ixodes scapularis is well established across much of the Northeastern and Mid-Atlantic States, and our results are generally consistent with previous studies conducted near the areas we sampled. Newly established I. scapularis populations were documented in two locations: Washington, D.C. (Rock Creek Park) and Greene County, Virginia (Shenandoah National Park). This research demonstrates the potential risk of tick-borne pathogen exposure in national parks and can be used to educate park visitors about the importance of preventative actions to minimize tick exposure.

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Section: Methodsmentioning

confidence: 99%

Prevalence and Diversity of Tick-Borne Pathogens in NymphalIxodes scapularis(Acari: Ixodidae) in Eastern National Parks

Johnson¹,

Graham²,

Boegler³

et al. 2016

J Med Entomol

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“…Borrelia miyamotoi -positive ticks were subjected to amplification and sequencing of the B. miyamotoi gene encoding ATP-dependent Clp protease subunit A ( clpA ) using the protocol described on pubMLST (http://pubmlst.org/borrelia/; Margos et al, 2015) with modifications. Each 50-μL PCR contained 5–15 μl template and 25 μl HotStar Taq Master Mix, 500 nM each primer (BmclpAF1268 and BmclpAR2051; Table 1), and MgCl 2 at a final concentration of 2.0 mM.…”

Section: Methodsmentioning

confidence: 99%

“… CalRd610: CalFluor Red 610; Quas705: Quasar 705; BHQ2 and BHQ3: Black Hole Quencher 2 and 3 respectively. a glpQ primers and probe adapted from Bacon et al (2005). b http://pubmlst.org/borrelia; Margos et al, 2015. …”

Section: Tablementioning

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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“…MK945865; and MK945867). The selected seven samples were included in a PCR reaction using specific primer sets for the amplification of nifS, clpA, rplB, pyrG, recG, clpX, pepX and uvrA gene fragments for each sample, as previously described (Table 2) [35] with slight modifications. The amplification of targets was performed using GoTaq ® G2 Flexi DNA Polymerase kit (Promega).…”

Section: Detection Of Borrelia Spp By Nested Pcr Sequencing and Phymentioning

confidence: 99%

Borrelia miyamotoi and Borrelia burgdorferi (sensu lato) identification and survey of tick-borne encephalitis virus in ticks from north-eastern Germany

et al. 2020

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Background: Ixodes ricinus is the most common tick species in Europe and the main vector for Borrelia burgdorferi (sensu lato) and tick-borne encephalitis virus (TBEV). It is involved also in the transmission of Borrelia miyamotoi, a relapsing fever spirochete that causes health disorders in humans. Little is known regarding the circulation of Borrelia species and the natural foci of TBEV in northeastern Germany. The goal of this study was to investigate the infection rates of Borrelia spp. and of TBEV in I. ricinus ticks from northeastern Germany. Methods: Ticks were collected by flagging from 14 forest sites in Mecklenburg-Western Pomerania between April and October 2018. RNA and DNA extraction was performed from individual adult ticks and from pools of 2-10 nymphs. Real time reverse transcription PCR (RT-qPCR) targeted the 3′ non-coding region of TBEV, while DNA of Borrelia spp. was tested by nested PCR for the amplification of 16S-23S intergenic spacer. Multilocus sequence typing (MLST) was performed on B. miyamotoi isolates. Results: In total, 2407 ticks were collected (239 females, 232 males and 1936 nymphs). Female and male I. ricinus ticks had identical infection rates (both 12.1%) for Borrelia spp., while nymphal pools showed a minimum infection rate (MIR) of 3.3%. Sequencing revealed four Borrelia species: B. afzelii, B. garinii, B. valaisiana and B. miyamotoi. Borrelia afzelii had the highest prevalence in adult ticks (5.5%) and nymphs (MIR of 1.8%). Borrelia miyamotoi was identified in 3.0% of adults and registered the MIR of 0.8% in nymphs. Borrelia valaisiana was confirmed in 2.5% adult ticks and nymphs had the MIR of 0.7%, while B. garinii was present in 1.1% of adults and showed a MIR of 0.1% in nymphs. The MLST of B. miyamotoi isolates showed that they belong to sequence type 635. No tick sample was positive after RT-qPCR for TBEV RNA. Conclusions: The prevalence of B. miyamotoi in I. ricinus ticks registered similar levels to other reports from Europe suggesting that this agent might be well established in the local tick population. The detection of B. burgdorferi (s.l.) indicates a constant circulation in tick populations from this region.

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PubMLST.org – The new home for the Borrelia MLSA database

Cited by 26 publications

References 11 publications

Prevalence and Diversity of Tick-Borne Pathogens in NymphalIxodes scapularis(Acari: Ixodidae) in Eastern National Parks

Prevalence and Diversity of Tick-Borne Pathogens in NymphalIxodes scapularis(Acari: Ixodidae) in Eastern National Parks

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

Borrelia miyamotoi and Borrelia burgdorferi (sensu lato) identification and survey of tick-borne encephalitis virus in ticks from north-eastern Germany

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