European Borrelia burgdorferi isolated from humans and ticks culture conditions and antibiotic susceptibility

Preac-Mursic, V.; Wilske, Bettina; Schierz, G

doi:10.1016/s0176-6724(86)80110-9

Cited by 195 publications

(192 citation statements)

References 6 publications

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“…Citrated blood samples were transported to the laboratory where they were processed, and only plasma was inoculated in modified Kelly-Pettenkofer medium (34). All the samples were incubated at 33°C and checked for the presence of borreliae weekly (27). Isolated strains were grown in the same medium for two to four subcultures.…”

Section: Methodsmentioning

confidence: 99%

“…Isolation of the etiological agent from patient material is the most reliable method for the diagnosis of borrelial infection (1,14,16,23,26,27,47). In addition, it also provides live microorganisms; data obtained from their further characterization provide potentially valuable information on the geographical distribution, epidemiology, and pathogenesis of borrelial infection and Lyme borreliosis.…”

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confidence: 99%

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Comparison of Borrelia burgdorferi Sensu Lato Strains Isolated from Specimens Obtained Simultaneously from Two Different Sites of Infection in Individual Patients

et al. 2005

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The aim of the present study was to analyze and compare Borrelia strains isolated from two different specimens obtained simultaneously from individual patients with Lyme borreliosis. Fifty such patients and 50 corresponding pairs of Borrelia isolates (100 low-propagated strains) were subjected to genotypic and phenotypic analysis, including pulsed-field gel electrophoresis for species identification and plasmid profile determination and protein profile electrophoresis for the assessment of the presence and molecular masses of separated proteins. The strains were isolated from two distinct skin lesions (12 patients), skin and blood (28 patients), skin and cerebrospinal fluid (8 patients), and blood and cerebrospinal fluid (2 patients). Out of 100 isolates, 63 were typed as B. afzelii and 37 as B. garinii. From each individual specimen only a single Borrelia species was cultured. Comparison of 50 Borrelia strain pairs isolated from two different specimens of an individual patient revealed that 12/50 (24%) patients were simultaneously infected with two different Borrelia strains; in 3/50 (6%) patients strains differed at the species level, in 4 out of the remaining 47 (9%) patients a strain difference in plasmid profile was established, while 5 out of the remaining 43 (11%) patient strain pairs differed in regard to the protein profiles of the two concurrently isolated strains. The results of the present study indicate that human patients with Lyme borreliosis may simultaneously harbor different B. burgdorferi sensu lato strains.

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Comparison of Borrelia burgdorferi Sensu Lato Strains Isolated from Specimens Obtained Simultaneously from Two Different Sites of Infection in Individual Patients

et al. 2005

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“…(i) Culture techniques. The liquid media currently used to grow B. burgdorferi sensu lato were derived from the original Kelly medium (152) through various modifications made over time (17,19,267,328,329 [267]) are better able to support growth of B. burgdorferi sensu lato in terms of recovery from low inocula, shorter generation times of the spirochete, and maximal concentration of spirochetes in culture (ϳ10 8 to 10 9 / ml) ( Table 1). Key ingredients of BSK II include CMRL-1066, which is a standard medium used for growing various types of mammalian cells; bovine serum albumin fraction V, which serves as a rich source of protein and to stabilize the pH; N-acetylglucosamine, a precursor for bacterial cell wall biosynthesis; rabbit serum; citrate; pyruvate; and many others.…”

Section: Laboratory Diagnosismentioning

confidence: 99%

Diagnosis of Lyme Borreliosis

et al. 2005

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A large amount of knowledge has been acquired since the original descriptions of Lyme borreliosis (LB) and of its causative agent, Borrelia burgdorferi sensu stricto. The complexity of the organism and the variations in the clinical manifestations of LB caused by the different B. burgdorferi sensu lato species were not then anticipated. Considerable improvement has been achieved in detection of B. burgdorferi sensu lato by culture, particularly of blood specimens during early stages of disease. Culturing plasma and increasing the volume of material cultured have accomplished this. Further improvements might be obtained if molecular methods are used for detection of growth in culture and if culture methods are automated. Unfortunately, culture is insensitive in extracutaneous manifestations of LB. PCR and culture have high sensitivity on skin samples of patients with EM whose diagnosis is based mostly on clinical recognition of the lesion. PCR on material obtained from extracutaneous sites is in general of low sensitivity, with the exception of synovial fluid. PCR on synovial fluid has shown a sensitivity of up to >90% (when using four different primer sets) in patients with untreated or partially treated Lyme arthritis, making it a helpful confirmatory test in these patients. Currently, the best use of PCR is for confirmation of the clinical diagnosis of suspected Lyme arthritis in patients who are IgG immunoblot positive. PCR should not be used as the sole laboratory modality to support a clinical diagnosis of extracutaneous LB. PCR positivity in seronegative patients suspected of having late manifestations of LB most likely represents a false-positive result. Because of difficulties in direct methods of detection, laboratory tests currently in use are mainly those detecting antibodies to B. burgdorferi sensu lato. Tests used to detect antibodies to B. burgdorferi sensu lato have evolved from the initial formats as more knowledge on the immunodominant antigens has been collected. The recommendation for two-tier testing was an attempt to standardize testing and improve specificity in the United States. First-tier assays using whole-cell sonicates of B. burgdorferi sensu lato need to be standardized in terms of antigen composition and detection threshold of specific immunoglobulin classes. The search for improved serologic tests has stimulated the development of recombinant protein antigens and the synthesis of specific peptides from immunodominant antigens. The use of these materials alone or in combination as the source of antigen in a single-tier immunoassay may someday replace the currently recommended two-tier testing strategy. Evaluation of these assays is currently being done, and there is evidence that certain of these antigens may be broadly cross-reactive with the B. burgdorferi sensu lato species causing LB in Europe

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“…Spirochaetes were grown in 10 ml of modi®ed BarbourStoenner-Kelly (BSK) medium [14] at 338C for 5±6 days up to a cell density of c. 10 7 /ml, dispensed into 1.8-ml screw-capped tubes, and stored at À708C until used.…”

Section: B Afzelii Isolates and Culture Conditionsmentioning

confidence: 99%

Borreliacidal activity of early Lyme disease sera against complement-resistant Borrelia afzelii FEM1 wild-type and an OspC-lacking FEM1 variant

Kraiczy¹,

Hunfeld

Peters³

et al. 2000

Journal of Medical Microbiology

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Sera obtained from 14 Lyme borreliosis patients at early stages (stages I and II) of the disease were examined for their borreliacidal properties against Borrelia afzelii isolate FEM1 by use of a growth inhibition assay. Five of 14 immune sera exhibited borreliacidal activity against isolate FEM1. Heat-inactivated immune sera failed to kill the spirochaetes. Immunoblotting experiments with outer-membrane preparations showed that OspC and 11 additional proteins of 14.0, 16.0, 17.7, 19.3, 21.7, 27.5, 32.7, 40.7, 48.9, 51.3 and 53.6 kDa were recognised by borreliacidal immune sera. To analyse the borreliacidal properties of anti-OspC antibodies, two sera (EM4 and EM5), which beside antibodies against a 51.3-kDa protein contained exclusively anti-OspC antibodies, were further investigated by comparative analysis with a FEM1 wild-type and a FEM1 variant lacking OspC in a growth inhibition assay. Only FEM1 wild-type and not variant FEM1OspC(2) was killed by immune sera EM4 and EM5. Complementdependent killing of FEM1 wild-type was mediated by formation of the terminal complement complex that was found to be attached directly to the outer membrane as con®rmed by immuno-electron microscopy. No complement deposition was observed on the surface of variant FEM1OspC(2) after incubation with immune sera EM4 and EM5, thereby suggesting that only anti-OspC antibodies in these two immune sera were responsible for borreliacidal activity. These results provide direct evidence that antiOspC antibodies, once developed during the immune response, are of critical importance for ef®cient killing of borreliae in the early phase of infection.

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European Borrelia burgdorferi isolated from humans and ticks culture conditions and antibiotic susceptibility

Cited by 195 publications

References 6 publications

Comparison of Borrelia burgdorferi Sensu Lato Strains Isolated from Specimens Obtained Simultaneously from Two Different Sites of Infection in Individual Patients

Comparison of Borrelia burgdorferi Sensu Lato Strains Isolated from Specimens Obtained Simultaneously from Two Different Sites of Infection in Individual Patients

Diagnosis of Lyme Borreliosis

Borreliacidal activity of early Lyme disease sera against complement-resistant Borrelia afzelii FEM1 wild-type and an OspC-lacking FEM1 variant

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