Quantitation of cell-associated borrelial DNA in the blood of Lyme disease patients with erythema migrans

Liveris, Dionysios; Schwartz, Ira; McKenna, Donna; Nowakowski, John; Nadelman, Robert B.; DeMarco, Joseph P.; Iyer, Radha; Cox, Mary E.; Holmgren, Diane; Wormser, Gary P.

doi:10.1007/s10096-011-1376-x

Cited by 26 publications

(33 citation statements)

References 17 publications

(23 reference statements)

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“…Epidemiological studies often involve isolation and characterization of multiple B. burgdorferi strains from regions of endemicity (58,59,66,67,102). However, there is no simple test to determine whether a new isolate is indeed a very different strain or is similar to one of the strains previously isolated from the same region.…”

Section: Discussionmentioning

confidence: 99%

Detection of Established Virulence Genes and Plasmids To Differentiate Borrelia burgdorferi Strains

2012

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ABSTRACTBorrelia burgdorferisensu stricto is the major causative agent of Lyme disease in the United States, whileB. gariniiandB. afzeliiare more prevalent in Europe. The highly complex genome ofB. burgdorferiis comprised of a linear chromosome and a large number of variably sized linear and circular plasmids. Many plasmids of this spirochete are unstable during its culturein vitro. Given that many of theB. burgdorferivirulence factors identified to date are plasmid encoded, spirochetal plasmid content determination is essential for genetic analysis of Lyme pathogenesis. Although PCR-based assays facilitate plasmid profiling of sequencedB. burgdorferistrains, a rapid genetic content determination strategy for nonsequenced strains has not yet been described. In this study, we combined pulsed-field gel electrophoresis (PFGE) and Southern hybridization for detection of genes encoding known virulence factors,ribosomal RNA genespacer restriction fragment length polymorphismtypes (RSTs),ospCgroup determination, and sequencing of the variabledbpAandospCgenes. We show that two strains isolated from the same tick and both originally named N40 are in fact very distinct. Furthermore, we failed to detectbbk32, which encodes a fibronectin-binding adhesin, in one “N40” strain. Thus, two distinct strains that show different plasmid profiles, as determined by PFGE and PCR, were isolated from the same tick and vary in theirospCanddbpAsequences. However, both belong to group RST3B.

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Section: Discussionmentioning

confidence: 99%

Detection of Established Virulence Genes and Plasmids To Differentiate Borrelia burgdorferi Strains

2012

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“…Samples from patients with early localized (only an EM) and early disseminated (Lyme carditis or Lyme neuroborreliosis) Lyme disease were tested by culture and PCR when possible. Culture and PCR were performed according to published methods (8)(9)(10)(11). Additionally, joint fluid from patients with late disseminated Lyme arthritis was tested by PCR using the published methods of Li et al (12).…”

Section: Methodsmentioning

confidence: 99%

Collection and Characterization of Samples for Establishment of a Serum Repository for Lyme Disease Diagnostic Test Development and Evaluation

et al. 2014

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Serological assays and a two-tiered test algorithm are recommended for laboratory confirmation of Lyme disease. In the United States, the sensitivity of two-tiered testing using commercially available serology-based assays is dependent on the stage of infection and ranges from 30% in the early localized disease stage to near 100% in late-stage disease. Other variables, including subjectivity in reading Western blots, compliance with two-tiered recommendations, use of different first-and second-tier test combinations, and use of different test samples, all contribute to variation in two-tiered test performance. The availability and use of sample sets from well-characterized Lyme disease patients and controls are needed to better assess the performance of existing tests and for development of improved assays. To address this need, the Centers for Disease Control and Prevention and the National Institutes of Health prospectively collected sera from patients at all stages of Lyme disease, as well as healthy donors and patients with look-alike diseases. Patients and healthy controls were recruited using strict inclusion and exclusion criteria. Samples from all included patients were retrospectively characterized by two-tiered testing. The results from two-tiered testing corroborated the need for novel and improved diagnostics, particularly for laboratory diagnosis of earlier stages of infection. Furthermore, the two-tiered results provide a baseline with samples from well-characterized patients that can be used in comparing the sensitivity and specificity of novel diagnostics. Panels of sera and accompanying clinical and laboratory testing results are now available to Lyme disease serological test users and researchers developing novel tests.

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“…In other spirochetes, the number of infecting bacteria in whole blood samples were found within the range of 10 4 to 10 5 bacteria/mL. 28,29 In human TPA infections, a reported number of bacteria/mL of whole blood reached 1 × 10 3 and 3 × 10 4 . 30,31 The total blood volume (~5 L) therefore likely contains in one point of TPA infection at least 10 8 treponemes.…”

Section: Macrolide Failures In Syphilis Treatment and Frequency Of Mumentioning

confidence: 99%

Macrolide Resistance in the Syphilis Spirochete, Treponema pallidum ssp. pallidum: Can We Also Expect Macrolide-Resistant Yaws Strains?

Šmajs

Paštěková

Grillová

2015

The American Society of Tropical Medicine and Hygiene

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Abstract. Treponema pallidum ssp. pallidum (TPA) causes over 10 million new cases of syphilis worldwide whereas T. pallidum ssp. pertenue (TPE), the causative agent of yaws, affects about 2.5 million people. Although penicillin remains the drug of choice in the treatment of syphilis, in penicillin-allergic patients, macrolides have been used in this indication since the 1950s. Failures of macrolides in syphilis treatment have been well documented in the literature and since 2000, there has been a dramatic increase in a number of clinical samples with macrolide-resistant TPA. Scarce data regarding the genetics of macrolide-resistant mutations in TPA suggest that although macrolide-resistance mutations have emerged independently several times, the increase in the proportion of TPA strains resistant to macrolides is mainly due to the spread of resistant strains, especially in developed countries. The emergence of macrolide resistance in TPA appears to require a two-step process including either A2058G or A2059G mutation in one copy of the 23S rRNA gene and a subsequent gene conversion unification of both rRNA genes. Given the enormous genetic similarity that was recently revealed between TPA and TPE strains, there is a low but reasonable risk of emergence and spread of macrolide-resistant yaws strains following azithromycin treatment.

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Quantitation of cell-associated borrelial DNA in the blood of Lyme disease patients with erythema migrans

Cited by 26 publications

References 17 publications

Detection of Established Virulence Genes and Plasmids To Differentiate Borrelia burgdorferi Strains

Detection of Established Virulence Genes and Plasmids To Differentiate Borrelia burgdorferi Strains

Collection and Characterization of Samples for Establishment of a Serum Repository for Lyme Disease Diagnostic Test Development and Evaluation

Macrolide Resistance in the Syphilis Spirochete, Treponema pallidum ssp. pallidum: Can We Also Expect Macrolide-Resistant Yaws Strains?

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