Genotypic Variation and Mixtures of Lyme Borrelia in Ixodes Ticks from North America and Europe

Crowder, Chris D.; Matthews, Heather; Schutzer, Steven E.; Rounds, Megan A.; Luft, Benjamin J.; Nolte, Oliver; Campbell, Scott R.; Phillipson, Curtis A.; Li, Feng; Sampath, Ranga; Ecker, David J.; Eshoo, Mark W.

doi:10.1371/journal.pone.0010650

Cited by 85 publications

(80 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Enrichment of a negative control was also performed. Following isothermal Borrelia DNA enrichment, a broad-range PCR followed by electrospray ionization mass spectrometry (IA/PCR/ ESI-MS) was performed, in which each sample was subjected to a series of eight diagnostic PCRs (32,33). By the selection of Borrelia PCR targets that vary in sequence between Borrelia species and strains (genotypes), it is possible to use the IA/PCR/ESI-MS technology to identify the Borrelia species and distinguish its genotype even when present in mixtures of genotypes (32,33).…”

Section: Methodsmentioning

confidence: 99%

“…Following isothermal Borrelia DNA enrichment, a broad-range PCR followed by electrospray ionization mass spectrometry (IA/PCR/ ESI-MS) was performed, in which each sample was subjected to a series of eight diagnostic PCRs (32,33). By the selection of Borrelia PCR targets that vary in sequence between Borrelia species and strains (genotypes), it is possible to use the IA/PCR/ESI-MS technology to identify the Borrelia species and distinguish its genotype even when present in mixtures of genotypes (32,33). A volume of 10 l of isothermal amplified nucleic acid extract was used in each PCR, and all PCRs were analyzed using an electrospray ionization mass spectrometry system (Abbott Molecular, Des Plaines, IL) as previously described (32,33).…”

Section: Methodsmentioning

confidence: 99%

“…By the selection of Borrelia PCR targets that vary in sequence between Borrelia species and strains (genotypes), it is possible to use the IA/PCR/ESI-MS technology to identify the Borrelia species and distinguish its genotype even when present in mixtures of genotypes (32,33). A volume of 10 l of isothermal amplified nucleic acid extract was used in each PCR, and all PCRs were analyzed using an electrospray ionization mass spectrometry system (Abbott Molecular, Des Plaines, IL) as previously described (32,33). In silico analysis.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Borrelia burgdorferi Not Confirmed in Human-Biting Amblyomma americanum Ticks from the Southeastern United States

et al. 2015

Self Cite

View full text Add to dashboard Cite

The predominant human-biting tick throughout the southeastern United States is Amblyomma americanum. Its ability to transmit pathogens causing Lyme disease-like illnesses is a subject of ongoing controversy. Results of previous testing by the Department of Defense Human Tick Test Kit Program and other laboratories indicated that it is highly unlikely that A. americanum transmits any pathogen that causes Lyme disease. In contrast, a recent publication by Clark and colleagues (K. L. Clark, B. Leydet, and S. Hartman, Int. J. Med. Sci. 10:915-931, 2013) reported detection of Lyme group Borrelia in A. americanum using a nested-flagellin-gene PCR. We evaluated this assay by using it and other assays to test 1,097 A. americanum ticks collected from humans. Using the Clark assay, in most samples we observed nonspecific amplification and nonrepeatability of results on subsequent testing of samples. Lack of reaction specificity and repeatability is consistent with mispriming, likely due to high primer concentrations and low annealing temperatures in this protocol. In six suspect-positive samples, Borrelia lonestari was identified by sequencing of an independent gene region; this is not a Lyme group spirochete and is not considered zoonotic. B. burgdorferi was weakly amplified from one pool using some assays, but not others, and attempts to sequence the amplicon of this pool failed, as did attempts to amplify and sequence B. burgdorferi from the five individual samples comprising this pool. Therefore, B. burgdorferi was not confirmed in any sample. Our results do not support the hypothesis that A. americanum ticks are a vector for Lyme group Borrelia infections. T he vectors and etiologic agents of Lyme-like diseases in the southeastern United States are a subject of ongoing controversy (1, 2). In the United States, most Lyme disease is caused by infection with Borrelia burgdorferi sensu stricto, a bacterium that is phylogenetically within the B. burgdorferi sensu lato "Lyme group" of spirochetes vectored by hard ticks. The Lyme group also includes genospecies implicated as the etiologic agents of Lyme disease in other geographic regions, including B. garinii, B. afzelii, B. spielmanii, and B. valaisiana in Europe. Additional Lyme group genospecies continue to be described. In the United States, these include B. americana, B. andersonii, and B. carolinensis, all of unknown pathogenicity, and B. bissettii, which has been implicated in cases of human illness (1). In contrast to the Lyme group, the relapsing fever (RF) group spirochetes, many of which are vectored by soft ticks, are a separate phylogenetic cluster that includes agents associated with RF disease in humans. In the United States, genospecies within the RF group include B. hermsii, B. turicatae, and B. parkeri, vectored by soft ticks, and B. miyamotoi, B. davisii, and B. lonestari, vectored by hard ticks. B. hermsii is the main cause of tick-borne relapsing fever in the United States; B. turicatae and B. parkeri have also been associated with human disease (3). B...

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Borrelia burgdorferi Not Confirmed in Human-Biting Amblyomma americanum Ticks from the Southeastern United States

et al. 2015

Self Cite

View full text Add to dashboard Cite

show abstract

“…associated with human infections, including unculturable organisms, directly from blood specimens (21)(22)(23)(24)(25)(26)(27). The assay can also detect three classes of antibiotic resistance markers associated with resistance to methicillin (mecA), vancomycin (vanA and vanB), and carbapenems (bla KPC ).…”

mentioning

confidence: 99%

Improved Sensitivity for Molecular Detection of Bacterial and Candida Infections in Blood

et al. 2014

Self Cite

View full text Add to dashboard Cite

The rapid identification of bacteria and fungi directly from the blood of patients with suspected bloodstream infections aids in diagnosis and guides treatment decisions. The development of an automated, rapid, and sensitive molecular technology capable of detecting the diverse agents of such infections at low titers has been challenging, due in part to the high background of genomic DNA in blood. PCR followed by electrospray ionization mass spectrometry (PCR/ESI-MS) allows for the rapid and accurate identification of microorganisms but with a sensitivity of about 50% compared to that of culture when using 1-ml wholeblood specimens. Here, we describe a new integrated specimen preparation technology that substantially improves the sensitivity of PCR/ESI-MS analysis. An efficient lysis method and automated DNA purification system were designed for processing 5 ml of whole blood. In addition, PCR amplification formulations were optimized to tolerate high levels of human DNA. An analysis of 331 specimens collected from patients with suspected bloodstream infections resulted in 35 PCR/ESI-MS-positive specimens (10.6%) compared to 18 positive by culture (5.4%). PCR/ESI-MS was 83% sensitive and 94% specific compared to culture. Replicate PCR/ESI-MS testing from a second aliquot of the PCR/ESI-MS-positive/culture-negative specimens corroborated the initial findings in most cases, resulting in increased sensitivity (91%) and specificity (99%) when confirmed detections were considered true positives. The integrated solution described here has the potential to provide rapid detection and identification of organisms responsible for bloodstream infections.

show abstract

“…Prior to this publication, only B. burgdorferi had been reported in U.S. patients without a history of travel to Europe or Asia, where B. garinii and B. afzelii are endemic (2)(3)(4). B. afzelii has not been isolated from nature in the Western Hemisphere; B. garinii has been detected in this hemisphere only in ticks infesting sea birds on coastal islands (Newfoundland) remote from human habitation (5).…”

mentioning

confidence: 99%

Assessment of New Culture Method for Detection of Borrelia Species from Serum of Lyme Disease Patients

2014

View full text Add to dashboard Cite

A novel method of culturing spirochetes from the serum of U.S. Lyme disease patients was recently reported by Sapi and colleagues to have 94% sensitivity and 100% specificity for Borrelia species as assessed by microscopy and DNA sequence analysis of the pyrG gene (E. Sapi, N. Pabbati, A. Datar, E. M. Davies, A. Rattelle, and B. A. Kuo, Int. J. Med. Sci. 10:362-376, 2013). The majority of the spirochetes described were related by pyrG sequences to species of Borrelia previously undetected in North American patients without a reported history of travel to Europe or Asia. To better understand these unexpected findings, we determined pyrG sequences of the laboratory reference strains used by the investigators for method development and testing of culture medium. Eighty percent (41/51) of the reported patient-derived pyrG sequences were identical to one of the laboratory strains, and an additional 12% (6/51) differed by only a single nucleotide across a 603-bp region of the pyrG gene. Thus, false positivity due to laboratory contamination of patient samples cannot be ruled out, and further validation of the proposed novel culture method is required.

show abstract

Genotypic Variation and Mixtures of Lyme Borrelia in Ixodes Ticks from North America and Europe

Cited by 85 publications

References 40 publications

Borrelia burgdorferi Not Confirmed in Human-Biting Amblyomma americanum Ticks from the Southeastern United States

Borrelia burgdorferi Not Confirmed in Human-Biting Amblyomma americanum Ticks from the Southeastern United States

Improved Sensitivity for Molecular Detection of Bacterial and Candida Infections in Blood

Assessment of New Culture Method for Detection of Borrelia Species from Serum of Lyme Disease Patients

Contact Info

Product

Resources

About