Ear punch biopsy method for detection and isolation of Borrelia burgdorferi from rodents

Sinsky, Richard J.; Piesman, Joseph

doi:10.1128/jcm.27.8.1723-1727.1989

Cited by 212 publications

(83 citation statements)

References 21 publications

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“…The rate of infection was 95% within the tick colony, determined by dissection of ticks and identifying spirochetes by dark field microscopy. Infection of the mice was determined by ear punch biopsy as described previously (19).…”

Section: Methodsmentioning

confidence: 99%

The major histocompatibility complex-restricted response of recombinant inbred strains of mice to natural tick transmission of Borrelia burgdorferi.

Golde

Burkot

Sviat

et al. 1993

The Journal of Experimental Medicine

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SummaryThe causative agent of Lyme disease, Borrelia burgdorferi, is transmitted by ticks of the Ixodes ricinus complex. In this study, we report the antibody response of recombinant inbred strains of mice of the H-2, b, d, and k haplotypes, infected with/~ burgdorferi as a result of exposure to infected I. daramini. The patterns of antibody response assayed by Western blot analysis indicate significant major histocompatibility complex (MHC) restriction to bacterial antigens within the first 2 mo of infection in mice. Other bacterial antigens induce a significant response across the MHC haplotypes tested when assayed on the same bacterial strain used to transmit the infection, but do not crossreact with the same proteins derived from heterologous strains of/~ burgdorferi.No response to outer surface protein A was detected at any time during the 60-d period we analyzed this infection. A third group of bacterial antigens appear to generate a MHC-nonrestricted response, and this lack of restriction is maintained when assaying the crossreactivity of the response with other strains of/~ burgdo~ri. These proteins may provide more accurate diagnostic probes than those currently in use. Finally, there appears to be a significant difference in the expression of most bacterial antigens when the spirochete is cultured for many passages since the same strain of bacterium isolated from low-passage and high-passage preparations exhibit different banding patterns in Western blots when assayed with the same sera.

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Section: Methodsmentioning

confidence: 99%

The major histocompatibility complex-restricted response of recombinant inbred strains of mice to natural tick transmission of Borrelia burgdorferi.

Golde

Burkot

Sviat

et al. 1993

The Journal of Experimental Medicine

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“…Low-passage t3, burgdorferi297 (Steere et aL, 1983) was provided by Russell Johnson and low-passage B, burgdorferi N40 was obtained from Stephen Barthold. Virulence of both low-passage isolates was confirmed by their ability to induce carditis and arthritis in two-week-old C3H/HeJ mice following intradermal inoculation with 1 × 104 bacteria and by recovery from ear punches (Sinsky and Piesman~ 1989) incubated in BSK II medium (Barbour, 1984) containing rifampicin (50 l~g ml-~) and amphotericin B (2.5 I~g ml-1). Virulent strains were passaged no more than seven times before experimental manipulations.…”

Section: Bacterial Strains and Plasmidsmentioning

confidence: 99%

Evidence for in vivo but not in vitro expression of a Borrelia burgdorferi outer surface protein F (OspF) homologue

Akins¹,

Porcella²,

Popova

et al. 1995

Molecular Microbiology

148

220

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Protein export signals from the low-passage 297 strain of Borrelia burgdorferi were cloned as fusions with an Escherichia coli alkaline phosphatase (PhoA) reporter lacking a signal sequence. One PhoA+ clone (BbK2.10-PhoA) was derived from a borrelial lipoprotein. Although the polypeptide encoded by the full-length bbk2.10 gene had 76% similarity and 56% identity to outer surface protein F (OspF) from B. burgdorferi strain N40, antibodies directed against recombinant forms of the two proteins revealed that they were not cross-reactive. The nucleotide sequences of bbk2.10 and ospF from the N40 and 297 strains, respectively, were determined to confirm that the N40 and 297 strains each contained both genes. Southern blot analysis revealed that bbk2.10 is a single-copy gene and that the B. burgdorferi strain 297 and N40 genomes appeared to contain one other gene more closely related to ospF than bbk2.10. It was particularly noteworthy that ospF, but not bbk2.10, was expressed in vitro while B. burgdorferi-infected mice generated antibodies reactive with both lipoproteins. To help confirm that the BbK2.10-reactive antibodies produced by the B. burgdorferi-infected mice were specific for that protein, a second gene, bbk2.11, which hybridized with the ospF probe was cloned; the corresponding polypeptide reacted strongly with OspF antisera but failed to react with BbK2.10-specific antisera. Taken together, these data demonstrate that BbK2.10, BbK2.11, and OspF comprise a B. burgdorferi lipoprotein family and that at least one member (BbK2.10) appears to be expressed only during infection.

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“…The samples derived from wild rodents were cultured in the standard modified BSKII medium rather than the enriched medium suggested by Sinsky & Piesman (1989) for the isolation of B. burgdorferi from ear punch biopsies. However, standard BSKII medium has been used successfully to isolate B. burgdorferi from vertebrate tissues collected in North America and continental Europe (Berger et a l .…”

Section: Discussionmentioning

confidence: 99%

Problems of isolating Borrelia burgdorferi from ticks collected in United Kingdom foci of Lyme disease

Livesley

Carey

Gern

et al. 1994

Medical Vet Entomology

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Many isolates of Borrelia burgdorferi have been obtained from ticks and vertebrate tissues collected in North America and continental Europe but only one established culture of United Kingdom Borrelia burgdorferi has been recorded. In this paper we report the isolation of B. burgdorferi from one of 108 tick pools representing 733 ticks and eight-four tissue samples from twenty-six rodents collected in the U.K, and the subsequent failure to establish the isolate (from ticks collected in Fordingbridge) in culture. In contrast, using identical techniques and culture medium, B. burgdorferi was isolated from one of seven tick pools collected in Switzerland, and from a single pool of ticks collected in Slovakia, and both isolates were successfully passaged. Analysis of questing I. ricinus collected from Fordingbridge by direct immunofluorescence showed 6/32 (19%) of adults and 8/108 (7%) of nymphs were positive for B. burgdorferi, although only one nymph contained > or = 1000 spirochaetes. To examine further the problem of isolating U.K. B. burgdorferi, twelve Ixodes ricinus tick samples from Fordingbridge, a recognized focus of Lyme disease, were subjected to isolation and culturing techniques, and the procedures monitored by use of the polymerase chain reaction (PCR). Whereas 11/12 samples were PCR positive after 2 weeks in culture, only one was PCR positive after 4 weeks. Motile spirochaetes were not visible by dark-field microscopy in any of the cultures. The results indicate that the standard BSK II medium routinely used to isolate and culture B. burgdorferi does not readily support the replication of the Borrelia species endemic to the U.K.

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Ear punch biopsy method for detection and isolation of Borrelia burgdorferi from rodents

Cited by 212 publications

References 21 publications

The major histocompatibility complex-restricted response of recombinant inbred strains of mice to natural tick transmission of Borrelia burgdorferi.

The major histocompatibility complex-restricted response of recombinant inbred strains of mice to natural tick transmission of Borrelia burgdorferi.

Evidence for in vivo but not in vitro expression of a Borrelia burgdorferi outer surface protein F (OspF) homologue

Problems of isolating Borrelia burgdorferi from ticks collected in United Kingdom foci of Lyme disease

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