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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1995 Aug;33(8):1989–1996. doi: 10.1128/jcm.33.8.1989-1996.1995

Characterization of Finnish Borrelia burgdorferi sensu lato isolates by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and with monoclonal antibodies.

J Tuomi 1, L K Rantamäki 1, R Tanskanen 1, J Junttila 1
PMCID: PMC228322  PMID: 7559935

Abstract

Thirty-seven Borrelia burgdorferi strains, isolated in 1992 from Ixodes ricinus in Finland, were investigated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and by immunoblotting and indirect immunofluorescence assay (IFA) with five to nine monoclonal antibodies (MAbs). By SDS-PAGE results and reactivities to MAbs H3TS, J 8.3, I 17.3, and D6, the 37 isolates were assigned to the species B. burgdorferi sensu stricto (n = 7), Borrelia afzelii (n = 17), or Borrelia garinii (n = 13). Twenty more isolates examined only by IFA and with part of the MAbs were distributed as follows: 9 B. burgdorferi sensu stricto and 11 other species. Among 16 of 37 isolates displaying a SDS-PAGE patterns considered typical of that of B. garinii, 3 were negative by the test with MAb D6; the rest were positive. The three MAb D6-negative isolates reacted with MAb J 8.3 but not with MAb I 17.3. It is suggested that these isolates of a previously undescribed type represent atypical B. afzelii strains deficient in the expression of OspB proteins. The misleading species designation by the SDS-PAGE result is described. The IFA results were generally consistent with those obtained by immunoblotting. The exception was for 3 of 29 isolates that were positive with MAb H5332 by immunoblotting but that were IFA negative. In the present material of 57 strains, all 16 B. burgdorferi sensu stricto isolates originated from the Aland Islands. B. afzelii and B. garinii were isolated from all three regions where ticks were collected. The distributive difference seems to offer a basis for comparative clinico-epidemiological studies of Lyme borreliosis.

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Selected References

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  1. Adam T., Gassmann G. S., Rasiah C., Göbel U. B. Phenotypic and genotypic analysis of Borrelia burgdorferi isolates from various sources. Infect Immun. 1991 Aug;59(8):2579–2585. doi: 10.1128/iai.59.8.2579-2585.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Asbrink E. Cutaneous manifestations of Lyme borreliosis. Clinical definitions and differential diagnoses. Scand J Infect Dis Suppl. 1991;77:44–50. [PubMed] [Google Scholar]
  3. Asbrink E., Hovmark A., Hederstedt B. The spirochetal etiology of acrodermatitis chronica atrophicans Herxheimer. Acta Derm Venereol. 1984;64(6):506–512. [PubMed] [Google Scholar]
  4. Asbrink E., Hovmark A., Olsson I. Clinical manifestations of acrodermatitis chronica atrophicans in 50 Swedish patients. Zentralbl Bakteriol Mikrobiol Hyg A. 1986 Dec;263(1-2):253–261. doi: 10.1016/s0176-6724(86)80128-6. [DOI] [PubMed] [Google Scholar]
  5. Assous M. V., Postic D., Paul G., Névot P., Baranton G. Western blot analysis of sera from Lyme borreliosis patients according to the genomic species of the Borrelia strains used as antigens. Eur J Clin Microbiol Infect Dis. 1993 Apr;12(4):261–268. doi: 10.1007/BF01967256. [DOI] [PubMed] [Google Scholar]
  6. Baranton G., Postic D., Saint Girons I., Boerlin P., Piffaretti J. C., Assous M., Grimont P. A. Delineation of Borrelia burgdorferi sensu stricto, Borrelia garinii sp. nov., and group VS461 associated with Lyme borreliosis. Int J Syst Bacteriol. 1992 Jul;42(3):378–383. doi: 10.1099/00207713-42-3-378. [DOI] [PubMed] [Google Scholar]
  7. Barbour A. G., Hayes S. F., Heiland R. A., Schrumpf M. E., Tessier S. L. A Borrelia-specific monoclonal antibody binds to a flagellar epitope. Infect Immun. 1986 May;52(2):549–554. doi: 10.1128/iai.52.2.549-554.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. 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]
  11. 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]
  12. Barbour A. G., Tessier S. L., Todd W. J. Lyme disease spirochetes and ixodid tick spirochetes share a common surface antigenic determinant defined by a monoclonal antibody. Infect Immun. 1983 Aug;41(2):795–804. doi: 10.1128/iai.41.2.795-804.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Barthold S. W. Infectivity of Borrelia burgdorferi relative to route of inoculation and genotype in laboratory mice. J Infect Dis. 1991 Feb;163(2):419–420. doi: 10.1093/infdis/163.2.419. [DOI] [PubMed] [Google Scholar]
  14. Belfaiza J., Postic D., Bellenger E., Baranton G., Girons I. S. Genomic fingerprinting of Borrelia burgdorferi sensu lato by pulsed-field gel electrophoresis. J Clin Microbiol. 1993 Nov;31(11):2873–2877. doi: 10.1128/jcm.31.11.2873-2877.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Bergström S., Olsén B., Burman N., Gothefors L., Jaenson T. G., Jonsson M., Mejlon H. A. Molecular characterization of Borrelia burgdorferi isolated from Ixodes ricinus in northern Sweden. Scand J Infect Dis. 1992;24(2):181–188. doi: 10.3109/00365549209052610. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Boerlin P., Peter O., Bretz A. G., Postic D., Baranton G., Piffaretti J. C. Population genetic analysis of Borrelia burgdorferi isolates by multilocus enzyme electrophoresis. Infect Immun. 1992 Apr;60(4):1677–1683. doi: 10.1128/iai.60.4.1677-1683.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Bundoc V. G., Barbour A. G. Clonal polymorphisms of outer membrane protein OspB of Borrelia burgdorferi. Infect Immun. 1989 Sep;57(9):2733–2741. doi: 10.1128/iai.57.9.2733-2741.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Burgdorfer W., Lane R. S., Barbour A. G., Gresbrink R. A., Anderson J. R. The western black-legged tick, Ixodes pacificus: a vector of Borrelia burgdorferi. Am J Trop Med Hyg. 1985 Sep;34(5):925–930. doi: 10.4269/ajtmh.1985.34.925. [DOI] [PubMed] [Google Scholar]
  20. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  21. Canica M. M., Nato F., du Merle L., Mazie J. C., Baranton G., Postic D. Monoclonal antibodies for identification of Borrelia afzelii sp. nov. associated with late cutaneous manifestations of Lyme borreliosis. Scand J Infect Dis. 1993;25(4):441–448. doi: 10.3109/00365549309008525. [DOI] [PubMed] [Google Scholar]
  22. Coleman J. L., Rogers R. C., Benach J. L. Selection of an escape variant of Borrelia burgdorferi by use of bactericidal monoclonal antibodies to OspB. Infect Immun. 1992 Aug;60(8):3098–3104. doi: 10.1128/iai.60.8.3098-3104.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Dressler F., Ackermann R., Steere A. C. Antibody responses to the three genomic groups of Borrelia burgdorferi in European Lyme borreliosis. J Infect Dis. 1994 Feb;169(2):313–318. doi: 10.1093/infdis/169.2.313. [DOI] [PubMed] [Google Scholar]
  24. Eiffert H., Ohlenbusch A., Fehling W., Lotter H., Thomssen R. Nucleotide sequence of the ospAB operon of a Borrelia burgdorferi strain expressing OspA but not OspB. Infect Immun. 1992 May;60(5):1864–1868. doi: 10.1128/iai.60.5.1864-1868.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Herzer P. Joint manifestations of Lyme borreliosis in Europe. Scand J Infect Dis Suppl. 1991;77:55–63. [PubMed] [Google Scholar]
  26. Jobe D. A., Callister S. M., Schell R. F. Recovery of Borrelia burgdorferi by filtration. J Clin Microbiol. 1993 Jul;31(7):1896–1898. doi: 10.1128/jcm.31.7.1896-1898.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Junttila J., Tanskanen R., Tuomi J. Prevalence of Borrelia burgdorferi in selected tick populations in Finland. Scand J Infect Dis. 1994;26(3):349–355. doi: 10.3109/00365549409011805. [DOI] [PubMed] [Google Scholar]
  28. Kurashige S., Bissett M., Oshiro L. Characterization of a tick isolate of Borrelia burgdorferi that possesses a major low-molecular-weight surface protein. J Clin Microbiol. 1990 Jun;28(6):1362–1366. doi: 10.1128/jcm.28.6.1362-1366.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  30. Postic D., Edlinger C., Richaud C., Grimont F., Dufresne Y., Perolat P., Baranton G., Grimont P. A. Two genomic species in Borrelia burgdorferi. Res Microbiol. 1990 May;141(4):465–475. doi: 10.1016/0923-2508(90)90072-x. [DOI] [PubMed] [Google Scholar]
  31. Péter O., Bretz A. G. Polymorphism of outer surface proteins of Borrelia burgdorferi as a tool for classification. Zentralbl Bakteriol. 1992 Jun;277(1):28–33. doi: 10.1016/s0934-8840(11)80867-4. [DOI] [PubMed] [Google Scholar]
  32. Rosa P. A., Schwan T., Hogan D. Recombination between genes encoding major outer surface proteins A and B of Borrelia burgdorferi. Mol Microbiol. 1992 Oct;6(20):3031–3040. doi: 10.1111/j.1365-2958.1992.tb01761.x. [DOI] [PubMed] [Google Scholar]
  33. Sadziene A., Barbour A. G., Rosa P. A., Thomas D. D. An OspB mutant of Borrelia burgdorferi has reduced invasiveness in vitro and reduced infectivity in vivo. Infect Immun. 1993 Sep;61(9):3590–3596. doi: 10.1128/iai.61.9.3590-3596.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Schwan T. G., Burgdorfer W. Antigenic changes of Borrelia burgdorferi as a result of in vitro cultivation. J Infect Dis. 1987 Nov;156(5):852–853. doi: 10.1093/infdis/156.5.852-a. [DOI] [PubMed] [Google Scholar]
  35. Schwan T. G., Burgdorfer W., Garon C. F. Changes in infectivity and plasmid profile of the Lyme disease spirochete, Borrelia burgdorferi, as a result of in vitro cultivation. Infect Immun. 1988 Aug;56(8):1831–1836. doi: 10.1128/iai.56.8.1831-1836.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Shoberg R. J., Jonsson M., Sadziene A., Bergström S., Thomas D. D. Identification of a highly cross-reactive outer surface protein B epitope among diverse geographic isolates of Borrelia spp. causing Lyme disease. J Clin Microbiol. 1994 Feb;32(2):489–500. doi: 10.1128/jcm.32.2.489-500.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Steere A. C. Lyme disease. N Engl J Med. 1989 Aug 31;321(9):586–596. doi: 10.1056/NEJM198908313210906. [DOI] [PubMed] [Google Scholar]
  38. Theisen M., Frederiksen B., Lebech A. M., Vuust J., Hansen K. Polymorphism in ospC gene of Borrelia burgdorferi and immunoreactivity of OspC protein: implications for taxonomy and for use of OspC protein as a diagnostic antigen. J Clin Microbiol. 1993 Oct;31(10):2570–2576. doi: 10.1128/jcm.31.10.2570-2576.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wahlberg P., Granlund H., Nyman D., Panelius J., Seppälä I. Late Lyme borreliosis: epidemiology, diagnosis and clinical features. Ann Med. 1993 Aug;25(4):349–352. doi: 10.3109/07853899309147296. [DOI] [PubMed] [Google Scholar]
  40. Weber K., Schierz G., Wilske B., Preac-Mursic V. Zur Klinik und Atiologie der Acrodermatitis chronica atrophicans. Hautarzt. 1984 Nov;35(11):571–577. [PubMed] [Google Scholar]
  41. Wilske B., Preac-Mursic V., Göbel U. B., Graf B., Jauris S., Soutschek E., Schwab E., Zumstein G. An OspA serotyping system for Borrelia burgdorferi based on reactivity with monoclonal antibodies and OspA sequence analysis. J Clin Microbiol. 1993 Feb;31(2):340–350. doi: 10.1128/jcm.31.2.340-350.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Wilske B., Preac-Mursic V., Jauris S., Hofmann A., Pradel I., Soutschek E., Schwab E., Will G., Wanner G. Immunological and molecular polymorphisms of OspC, an immunodominant major outer surface protein of Borrelia burgdorferi. Infect Immun. 1993 May;61(5):2182–2191. doi: 10.1128/iai.61.5.2182-2191.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Wilske B., Preac-Mursic V., Schierz G., Busch K. V. Immunochemical and immunological analysis of European Borrelia burgdorferi strains. Zentralbl Bakteriol Mikrobiol Hyg A. 1986 Dec;263(1-2):92–102. doi: 10.1016/s0176-6724(86)80108-0. [DOI] [PubMed] [Google Scholar]
  44. Wilske B., Preac-Mursic V., Schierz G., Gueye W., Herzer P., Weber K. Immunochemische Analyse der Immunantwort bei Spätmanifestationen der Lyme Borreliose. Zentralbl Bakteriol Mikrobiol Hyg A. 1988 Mar;267(4):549–558. [PubMed] [Google Scholar]
  45. Wilske B., Preac-Mursic V., Schierz G., Kühbeck R., Barbour A. G., Kramer M. Antigenic variability of Borrelia burgdorferi. Ann N Y Acad Sci. 1988;539:126–143. doi: 10.1111/j.1749-6632.1988.tb31846.x. [DOI] [PubMed] [Google Scholar]

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