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. 1990 Mar;28(3):566–572. doi: 10.1128/jcm.28.3.566-572.1990

Detection of Borrelia burgdorferi infection in Ixodes dammini ticks with the polymerase chain reaction.

D H Persing 1, S R Telford 3rd 1, A Spielman 1, S W Barthold 1
PMCID: PMC269663  PMID: 1969867

Abstract

The polymerase chain reaction (PCR) was used to amplify DNA sequences of the etiologic agent of Lyme disease, Borrelia burgdorferi, and was applied to the detection of the spirochete in its tick vector. The target for PCR amplification was the OSP-A gene of strain B31; analysis of isolates from different geographical areas indicated that this gene could be used to identify most North American isolates. These methods were extended to the analysis of colony-derived and field-collected Ixodes dammini. OSP-A-specific sequences were identified in 15 of 15 colony-derived nymphal ticks that had fed previously on an infected animal; no such amplification products were detected in 8 control ticks. Segregated midgut tissues of field-collected adult and nymphal ticks from Nantucket Island, Mass., and the Crane Reserve, Ipswich, Mass., were examined by both direct fluorescent-antibody (DFA) staining and PCR. The DFA technique identified 16 infected ticks of 30 paired specimens; 15 of these specimens were positive by PCR. One specimen was positive by PCR that was DFA negative. Both live whole ticks and desiccated dead specimens were suitable for this analysis. Because only five ticks are suitable for DFA analysis, the use of PCR may extend the range of specimens that can be analyzed for the presence of the Lyme spirochete.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anderson J. F., Johnson R. C., Magnarelli L. A., Hyde F. W. Identification of endemic foci of Lyme disease: isolation of Borrelia burgdorferi from feral rodents and ticks (Dermacentor variabilis). J Clin Microbiol. 1985 Jul;22(1):36–38. doi: 10.1128/jcm.22.1.36-38.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson J. F., Johnson R. C., Magnarelli L. A., Hyde F. W., Myers J. E. Prevalence of Borrelia burgdorferi and Babesia microti in mice on islands inhabited by white-tailed deer. Appl Environ Microbiol. 1987 Apr;53(4):892–894. doi: 10.1128/aem.53.4.892-894.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Anderson J. F., Magnarelli L. A. Avian and mammalian hosts for spirochete-infected ticks and insects in a Lyme disease focus in Connecticut. Yale J Biol Med. 1984 Jul-Aug;57(4):627–641. [PMC free article] [PubMed] [Google Scholar]
  4. Anderson J. F., Magnarelli L. A., McAninch J. B. New Borrelia burgdorferi antigenic variant isolated from Ixodes dammini from upstate New York. J Clin Microbiol. 1988 Oct;26(10):2209–2212. doi: 10.1128/jcm.26.10.2209-2212.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Barbour A. G., Garon C. F. Linear plasmids of the bacterium Borrelia burgdorferi have covalently closed ends. Science. 1987 Jul 24;237(4813):409–411. doi: 10.1126/science.3603026. [DOI] [PubMed] [Google Scholar]
  6. Barbour A. G., Hayes S. F. Biology of Borrelia species. Microbiol Rev. 1986 Dec;50(4):381–400. doi: 10.1128/mr.50.4.381-400.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Barbour A. G., Heiland R. A., Howe T. R. Heterogeneity of major proteins in Lyme disease borreliae: a molecular analysis of North American and European isolates. J Infect Dis. 1985 Sep;152(3):478–484. doi: 10.1093/infdis/152.3.478. [DOI] [PubMed] [Google Scholar]
  8. Barbour A. G. Isolation and cultivation of Lyme disease spirochetes. Yale J Biol Med. 1984 Jul-Aug;57(4):521–525. [PMC free article] [PubMed] [Google Scholar]
  9. Barbour A. G., Schrumpf M. E. Polymorphisms of major surface proteins of Borrelia burgdorferi. Zentralbl Bakteriol Mikrobiol Hyg A. 1986 Dec;263(1-2):83–91. doi: 10.1016/s0176-6724(86)80107-9. [DOI] [PubMed] [Google Scholar]
  10. Barbour A. G., Tessier S. L., Hayes S. F. Variation in a major surface protein of Lyme disease spirochetes. Infect Immun. 1984 Jul;45(1):94–100. doi: 10.1128/iai.45.1.94-100.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Benach J. L., Bosler E. M., Hanrahan J. P., Coleman J. L., Habicht G. S., Bast T. F., Cameron D. J., Ziegler J. L., Barbour A. G., Burgdorfer W. Spirochetes isolated from the blood of two patients with Lyme disease. N Engl J Med. 1983 Mar 31;308(13):740–742. doi: 10.1056/NEJM198303313081302. [DOI] [PubMed] [Google Scholar]
  12. Bergström S., Bundoc V. G., Barbour A. G. Molecular analysis of linear plasmid-encoded major surface proteins, OspA and OspB, of the Lyme disease spirochaete Borrelia burgdorferi. Mol Microbiol. 1989 Apr;3(4):479–486. doi: 10.1111/j.1365-2958.1989.tb00194.x. [DOI] [PubMed] [Google Scholar]
  13. Bissett M. L., Hill W. Characterization of Borrelia burgdorferi strains isolated from Ixodes pacificus ticks in California. J Clin Microbiol. 1987 Dec;25(12):2296–2301. doi: 10.1128/jcm.25.12.2296-2301.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Burgdorfer W. Discovery of the Lyme disease spirochete and its relation to tick vectors. Yale J Biol Med. 1984 Jul-Aug;57(4):515–520. [PMC free article] [PubMed] [Google Scholar]
  15. DeLong E. F., Wickham G. S., Pace N. R. Phylogenetic stains: ribosomal RNA-based probes for the identification of single cells. Science. 1989 Mar 10;243(4896):1360–1363. doi: 10.1126/science.2466341. [DOI] [PubMed] [Google Scholar]
  16. Gassmann G. S., Kramer M., Göbel U. B., Wallich R. Nucleotide sequence of a gene encoding the Borrelia burgdorferi flagellin. Nucleic Acids Res. 1989 May 11;17(9):3590–3590. doi: 10.1093/nar/17.9.3590. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lane R. S., Burgdorfer W. Spirochetes in mammals and ticks (Acari: Ixodidae) from a focus of Lyme borreliosis in California. J Wildl Dis. 1988 Jan;24(1):1–9. doi: 10.7589/0090-3558-24.1.1. [DOI] [PubMed] [Google Scholar]
  18. Lastavica C. C., Wilson M. L., Berardi V. P., Spielman A., Deblinger R. D. Rapid emergence of a focal epidemic of Lyme disease in coastal Massachusetts. N Engl J Med. 1989 Jan 19;320(3):133–137. doi: 10.1056/NEJM198901193200301. [DOI] [PubMed] [Google Scholar]
  19. Mullis K. B., Faloona F. A. Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Methods Enzymol. 1987;155:335–350. doi: 10.1016/0076-6879(87)55023-6. [DOI] [PubMed] [Google Scholar]
  20. Ou C. Y., Kwok S., Mitchell S. W., Mack D. H., Sninsky J. J., Krebs J. W., Feorino P., Warfield D., Schochetman G. DNA amplification for direct detection of HIV-1 in DNA of peripheral blood mononuclear cells. Science. 1988 Jan 15;239(4837):295–297. doi: 10.1126/science.3336784. [DOI] [PubMed] [Google Scholar]
  21. Persing D. H., Landry M. L. In vitro amplification techniques for the detection of nucleic acids: new tools for the diagnostic laboratory. Yale J Biol Med. 1989 Mar-Apr;62(2):159–171. [PMC free article] [PubMed] [Google Scholar]
  22. Piesman J., Mather T. N., Dammin G. J., Telford S. R., 3rd, Lastavica C. C., Spielman A. Seasonal variation of transmission risk of Lyme disease and human babesiosis. Am J Epidemiol. 1987 Dec;126(6):1187–1189. doi: 10.1093/oxfordjournals.aje.a114757. [DOI] [PubMed] [Google Scholar]
  23. Piesman J., Mather T. N., Sinsky R. J., Spielman A. Duration of tick attachment and Borrelia burgdorferi transmission. J Clin Microbiol. 1987 Mar;25(3):557–558. doi: 10.1128/jcm.25.3.557-558.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Plasterk R. H., Simon M. I., Barbour A. G. Transposition of structural genes to an expression sequence on a linear plasmid causes antigenic variation in the bacterium Borrelia hermsii. Nature. 1985 Nov 21;318(6043):257–263. doi: 10.1038/318257a0. [DOI] [PubMed] [Google Scholar]
  25. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  26. Schwan T. G., Simpson W. J., Schrumpf M. E., Karstens R. H. Identification of Borrelia burgdorferi and B. hermsii using DNA hybridization probes. J Clin Microbiol. 1989 Aug;27(8):1734–1738. doi: 10.1128/jcm.27.8.1734-1738.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Spielman A., Wilson M. L., Levine J. F., Piesman J. Ecology of Ixodes dammini-borne human babesiosis and Lyme disease. Annu Rev Entomol. 1985;30:439–460. doi: 10.1146/annurev.en.30.010185.002255. [DOI] [PubMed] [Google Scholar]
  28. Steere A. C., Broderick T. F., Malawista S. E. Erythema chronicum migrans and Lyme arthritis: epidemiologic evidence for a tick vector. Am J Epidemiol. 1978 Oct;108(4):312–321. doi: 10.1093/oxfordjournals.aje.a112625. [DOI] [PubMed] [Google Scholar]
  29. Steere A. C., Grodzicki R. L., Kornblatt A. N., Craft J. E., Barbour A. G., Burgdorfer W., Schmid G. P., Johnson E., Malawista S. E. The spirochetal etiology of Lyme disease. N Engl J Med. 1983 Mar 31;308(13):733–740. doi: 10.1056/NEJM198303313081301. [DOI] [PubMed] [Google Scholar]
  30. Thole J. E., Hindersson P., de Bruyn J., Cremers F., van der Zee J., de Cock H., Tommassen J., van Eden W., van Embden J. D. Antigenic relatedness of a strongly immunogenic 65 kDA mycobacterial protein antigen with a similarly sized ubiquitous bacterial common antigen. Microb Pathog. 1988 Jan;4(1):71–83. doi: 10.1016/0882-4010(88)90049-6. [DOI] [PubMed] [Google Scholar]
  31. Wallis R. C., Brown S. E., Kloter K. O., Main A. J., Jr Erythema chronicum migrans and lyme arthritis: field study of ticks. Am J Epidemiol. 1978 Oct;108(4):322–327. doi: 10.1093/oxfordjournals.aje.a112626. [DOI] [PubMed] [Google Scholar]

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