Detection of Borrelia burgdorferi sensu lato in Ticks: Immunofluorescence Assay versus Polymerase Chain Reaction

Kahl, Olaf; Gern, Lise; Gray, Jeremy; Guy, Edward; Jongejan, Frans; Kirstein, F.; Kurtenbach, Klaus; Rijpkema, Sjoerd; Stanek, Gerold

doi:10.1016/s0934-8840(98)80122-9

Cited by 22 publications

(7 citation statements)

References 5 publications

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“…Nested PCR approaches are generally considered more sensitive than simple PCR approaches, but in practice they are doubtlessly more susceptible to contamination and may produce more false-positive results than a simple PCR. The detection of larger numbers of Borrelia-infected ticks by IFA than by PCR is a well-known phenomenon that was documented in a multilaboratory study (17). There, intralaboratory concordance between the two techniques ranged from 40 to 100%; in our study, the concordance between the IFA and the nested PCR was 92.8% and that between the IFA and the simple PCR according to Liebisch et al was 94.1% (30).…”

Section: Discussionsupporting

confidence: 58%

“…False-positive IFA results can sometimes be traced back to misidentification of single Borrelia-like structures showing fluorescence (17). False-negative PCR results, on the other hand, can be explained by inhibition of DNA amplification (40), which may be a particular problem with engorged ticks due to the presence of hemoglobin (7) but which also occurs with unfed ticks because of small amounts of heme that are retained during tick molt (15).…”

Section: Discussionmentioning

confidence: 99%

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Substantial Rise in the Prevalence of Lyme Borreliosis Spirochetes in a Region of Western Germany over a 10-Year Period

Kampen

Rötzel

Kurtenbach

et al. 2004

Appl Environ Microbiol

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More than a decade after a study on the transmission cycle of Borrelia burgdorferi sensu lato in the Siebengebirge, a nature reserve near Bonn, Germany, questing nymphal and adult Ixodes ricinus ticks were collected again in three selected areas of the same low mountain range and examined for infection with B. burgdorferi sensu lato. Between May and October 2001, a total of 1,754 ticks were collected by blanket dragging; 374 ticks were analyzed for B. burgdorferi sensu lato by both an immunofluorescence assay (IFA) and at least two different PCR tests, whereas 171 ticks were analyzed by PCR only. By combining all assays, an average of 14% of the ticks tested positive for B. burgdorferi sensu lato, 5.5, 15.8, and 21.8% in the three collection areas. Of the nymphs and adults examined, 12.9 and 21.1%, respectively, were found to be spirochete infected. A lower total infection prevalence was obtained by IFA (14.4%) than by a nested PCR approach (16.5%), but both were higher than that obtained by a simple PCR approach (11.9%). Compared with data collected over a decade ago, the mean infection prevalence of B. burgdorferi sensu lato in the ticks was significantly higher for all three biotopes, whereas a similar pattern of habitat-specific infection prevalence was observed. Genotyping of B. burgdorferi sensu lato revealed high relative prevalences of B. valaisiana (identified in 43.1% of infected ticks) and B. garinii (32.3%), whereas B. afzelii (12.3%) and B. burgdorferi sensu stricto (1.5%) were relatively rare. We conclude that B. burgdorferi sensu lato infection has increased in this region over the last 15 years due to presently unknown changes in ecological conditions, perhaps related to climate change or wildlife management.

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Section: Discussionsupporting

confidence: 58%

Section: Discussionmentioning

confidence: 99%

Substantial Rise in the Prevalence of Lyme Borreliosis Spirochetes in a Region of Western Germany over a 10-Year Period

Kampen

Rötzel

Kurtenbach

et al. 2004

Appl Environ Microbiol

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“…The BALB/c mice were infected with European strains of B. burgdorferi sensu lato by exposure to nymphal Ixodes ricinus ticks, as described previously (10,13). These ticks were field collected either in Malonne, Belgium, or Neuchâtel, Switzerland.…”

mentioning

confidence: 99%

C-Terminal Invariable Domain of VlsE Is Immunodominant but Its Antigenicity Is Scarcely Conserved among Strains of Lyme Disease Spirochetes

2001

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VlsE, the variable surface antigen of Borrelia burgdorferi, contains two invariable domains located at the amino and carboxyl terminal ends, respectively, and a central variable domain. In this study, both immunogenicity and antigenic conservation of the C-terminal invariable domain were assessed. Mouse antiserum to a 51-mer synthetic peptide (Ct) which reproduced the entire sequence of the C-terminal invariable domain of VlsE from B. burgdorferi strain B31 was reacted on immunoblots with whole-cell lysates extracted from spirochetes of 12 strains from the B. burgdorferi sensu lato species complex. The antiserum recognized only VlsE from strain B31, indicating that epitopes of this domain differed among these strains. When Ct was used as enzyme-linked immunosorbent assay (ELISA) antigen, all of the seven monkeys and six mice that were infected with B31 spirochetes produced a strong antibody response to this peptide, indicating that the C-terminal invariable domain is immunodominant. None of 12 monkeys and only 11 of 26 mice that were infected with strains other than B31 produced a detectable anti-Ct response, indicating a limited antigenic conservation of this domain among these strains. Twenty-six of 33 dogs that were experimentally infected by tick inoculation were positive by the Ct ELISA, while only 5 of 18 serum samples from dogs clinically diagnosed with Lyme disease contained detectable anti-Ct antibody. Fifty-seven of 64 serum specimens that were collected from American patients with Lyme disease were positive by the Ct ELISA, while only 12 of 21 European samples contained detectable anti-Ct antibody. In contrast, antibody to the more conserved invariable region IR 6 of VlsE was present in all of these dog and human serum samples.

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“…Despite our relatively low infection prevalence by PCR, Borrelia spirochetes were detected using an anti-ßagellin monoclonal antibody in 10.7% of ticks assayed in 2003. Disagreement between detection methods for B. burgdorferi has been reported (Kahl et al 1998, Gustafson et al 1989, and a higher B. burgdorferi infection rate in ticks was previously observed using IFA in comparison to PCR assay (Kahl et al 1998). Although the anti-ßagellin MAb H9724 is Borrelia-wide and will not recognize Treponema, Leptospira, or Spirochaeta, other Borrelia spp.…”

Section: Discussionmentioning

confidence: 98%

Disease Agents inAmblyomma americanumfrom Northeastern Georgia

Varela

Moore

Little³

2004

J Med Entomol

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Amblyomma americanum (lone star tick) is known or suspected to vector several organisms that are implicated as human pathogens, including Ehrlichia chaffeensis, E. ewingii, and Borrelia lonestari. These three agents have also been detected in white-tailed deer (Odocoileus virginianus). Because northeastern Georgia has a high abundance of both lone star ticks and white-tailed deer, and one of these organisms, E. chaffeensis, is already known to be endemic in the area, we assayed individual adult A. americanum, collected during the spring of 2001, 2002, and 2003, for these three organisms. A total of 400 ticks were dissected and tissues assayed by polymerase chain reaction (PCR) using Ehrlichia species-specific and Borrelia genus-wide primers. Of ticks tested, 2.0% (8/398) had evidence of E. chaffeensis, 4.8% (19/398) had evidence of E. ewingii, and 1.0% (4/398) had evidence of B. lonestari. Borrelia sp. spirochetes were also visualized by an indirect fluorescent antibody test, using an anti-flagellin monoclonal antibody (H9724), in a total of 10.7% (32/300) of ticks tested in 2003. These results reconfirm the presence of E. chaffeensis and establish evidence of E. ewingii and B. lonestari in questing adult A. americanum ticks from northeastern Georgia. Detection of at least two of the three organisms in ticks collected each year suggests that people in northeastern Georgia are at risk of infection with these organisms.

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Detection of Borrelia burgdorferi sensu lato in Ticks: Immunofluorescence Assay versus Polymerase Chain Reaction

Cited by 22 publications

References 5 publications

Substantial Rise in the Prevalence of Lyme Borreliosis Spirochetes in a Region of Western Germany over a 10-Year Period

Substantial Rise in the Prevalence of Lyme Borreliosis Spirochetes in a Region of Western Germany over a 10-Year Period

C-Terminal Invariable Domain of VlsE Is Immunodominant but Its Antigenicity Is Scarcely Conserved among Strains of Lyme Disease Spirochetes

Disease Agents inAmblyomma americanumfrom Northeastern Georgia

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