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. 1993 Dec;31(12):3115–3122. doi: 10.1128/jcm.31.12.3115-3122.1993

Comparative analysis of genetic variability among Borrelia burgdorferi isolates from Europe and the United States by restriction enzyme analysis, gene restriction fragment length polymorphism, and pulsed-field gel electrophoresis.

B C Zingg 1, J F Anderson 1, R C Johnson 1, R B LeFebvre 1
PMCID: PMC266361  PMID: 7905881

Abstract

The genomes of 62 North American and European Borrelia burgdorferi isolates were examined by restriction endonuclease analysis (REA), gene probe restriction fragment length polymorphism, and pulsed-field gel electrophoresis (PFGE). Hybridization of restriction fragments with the immunologically relevant 83-kDa antigen gene revealed polymorphisms and divided the isolates into three major groups. Group I included all but two of the American isolates and some of the European isolates. One of two Californian isolates (DN 127) and one Ixodes dammini isolate from New York (strain 25015), previously described as atypical, were distinct from the isolates in the three groups. Plasmid profile analysis and REA, the method with the highest level of discrimination, revealed extensive heterogeneity among isolates of the same major group. Our study demonstrates the usefulness of the 83-kDa antigen gene probe for dividing the isolates into major genogroups, whereas REA and plasmid profile analysis allow for a distinction of individual strains within these groups.

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Selected References

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

  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. Anderson J. F., Barthold S. W., Magnarelli L. A. Infectious but nonpathogenic isolate of Borrelia burgdorferi. J Clin Microbiol. 1990 Dec;28(12):2693–2699. doi: 10.1128/jcm.28.12.2693-2699.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Anderson J. F., Duray P. H., Magnarelli L. A. Borrelia burgdorferi and Ixodes dammini prevalent in the greater Philadelphia area. J Infect Dis. 1990 Apr;161(4):811–812. doi: 10.1093/infdis/161.4.811. [DOI] [PubMed] [Google Scholar]
  4. Anderson J. F., Duray P. H., Magnarelli L. A. Prevalence of Borrelia burgdorferi in white-footed mice and Ixodes dammini at Fort McCoy, Wis. J Clin Microbiol. 1987 Aug;25(8):1495–1497. doi: 10.1128/jcm.25.8.1495-1497.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Anderson J. F., Mintz E. D., Gadbaw J. J., Magnarelli L. A. Babesia microti, human babesiosis, and Borrelia burgdorferi in Connecticut. J Clin Microbiol. 1991 Dec;29(12):2779–2783. doi: 10.1128/jcm.29.12.2779-2783.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Baranton G., Perolat P., Dufresne Y., Postic D., Quentin R., Fouquet B. Isolement de Borrelia burgdorferi du liquide céphalorachidien d'un malade traité par la pénicilline. Presse Med. 1989 Mar 25;18(12):637–637. [PubMed] [Google Scholar]
  8. 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]
  9. Barbour A. G., Garon C. F. The genes encoding major surface proteins of Borrelia burgdorferi are located on a plasmid. Ann N Y Acad Sci. 1988;539:144–153. doi: 10.1111/j.1749-6632.1988.tb31847.x. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. 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]
  12. Barbour A. G. Plasmid analysis of Borrelia burgdorferi, the Lyme disease agent. J Clin Microbiol. 1988 Mar;26(3):475–478. doi: 10.1128/jcm.26.3.475-478.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Baril C., Richaud C., Baranton G., Saint Girons I. S. Linear chromosome of Borrelia burgdorferi. Res Microbiol. 1989 Oct;140(8):507–516. doi: 10.1016/0923-2508(89)90083-1. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Burgdorfer W., Barbour A. G., Hayes S. F., Benach J. L., Grunwaldt E., Davis J. P. Lyme disease-a tick-borne spirochetosis? Science. 1982 Jun 18;216(4552):1317–1319. doi: 10.1126/science.7043737. [DOI] [PubMed] [Google Scholar]
  16. Denny T. P., Gilmour M. N., Selander R. K. Genetic diversity and relationships of two pathovars of Pseudomonas syringae. J Gen Microbiol. 1988 Jul;134(7):1949–1960. doi: 10.1099/00221287-134-7-1949. [DOI] [PubMed] [Google Scholar]
  17. Dreiseikelmann B., Wackernagel W. Absence in Bacillus subtilis and Staphylococcus aureus of the sequence-specific deoxyribonucleic acid methylation that is conferred in Escherichia coli K-12 by the dam and dcm enzymes. J Bacteriol. 1981 Jul;147(1):259–261. doi: 10.1128/jb.147.1.259-261.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ferdows M. S., Barbour A. G. Megabase-sized linear DNA in the bacterium Borrelia burgdorferi, the Lyme disease agent. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5969–5973. doi: 10.1073/pnas.86.15.5969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Fikrig E., Barthold S. W., Persing D. H., Sun X., Kantor F. S., Flavell R. A. Borrelia burgdorferi strain 25015: characterization of outer surface protein A and vaccination against infection. J Immunol. 1992 Apr 1;148(7):2256–2260. [PubMed] [Google Scholar]
  20. Hinnebusch J., Bergström S., Barbour A. G. Cloning and sequence analysis of linear plasmid telomeres of the bacterium Borrelia burgdorferi. Mol Microbiol. 1990 May;4(5):811–820. doi: 10.1111/j.1365-2958.1990.tb00651.x. [DOI] [PubMed] [Google Scholar]
  21. Hughes C. A., Johnson R. C. Methylated DNA in Borrelia species. J Bacteriol. 1990 Nov;172(11):6602–6604. doi: 10.1128/jb.172.11.6602-6604.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hughes C. A., Kodner C. B., Johnson R. C. DNA analysis of Borrelia burgdorferi NCH-1, the first northcentral U.S. human Lyme disease isolate. J Clin Microbiol. 1992 Mar;30(3):698–703. doi: 10.1128/jcm.30.3.698-703.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. LeFebvre R. B., Lane R. S., Perng G. C., Brown J. A., Johnson R. C. DNA and protein analyses of tick-derived isolates of Borrelia burgdorferi from California. J Clin Microbiol. 1990 Apr;28(4):700–707. doi: 10.1128/jcm.28.4.700-707.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. LeFebvre R. B., Perng G. C., Johnson R. C. Characterization of Borrelia burgdorferi isolates by restriction endonuclease analysis and DNA hybridization. J Clin Microbiol. 1989 Apr;27(4):636–639. doi: 10.1128/jcm.27.4.636-639.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lefebvre R. B., Perng G. C., Johnson R. C. The 83-kilodalton antigen of Borrelia burgdorferi which stimulates immunoglobulin M (IgM) and IgG responses in infected hosts is expressed by a chromosomal gene. J Clin Microbiol. 1990 Jul;28(7):1673–1675. doi: 10.1128/jcm.28.7.1673-1675.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Marconi R. T., Garon C. F. Identification of a third genomic group of Borrelia burgdorferi through signature nucleotide analysis and 16S rRNA sequence determination. J Gen Microbiol. 1992 Mar;138(3):533–536. doi: 10.1099/00221287-138-3-533. [DOI] [PubMed] [Google Scholar]
  27. Mayer L. W. Use of plasmid profiles in epidemiologic surveillance of disease outbreaks and in tracing the transmission of antibiotic resistance. Clin Microbiol Rev. 1988 Apr;1(2):228–243. doi: 10.1128/cmr.1.2.228. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nelson M., McClelland M. Site-specific methylation: effect on DNA modification methyltransferases and restriction endonucleases. Nucleic Acids Res. 1991 Apr 25;19 (Suppl):2045–2071. doi: 10.1093/nar/19.suppl.2045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Oliver J. H., Jr, Owsley M. R., Hutcheson H. J., James A. M., Chen C., Irby W. S., Dotson E. M., McLain D. K. Conspecificity of the ticks Ixodes scapularis and I. dammini (Acari: Ixodidae). J Med Entomol. 1993 Jan;30(1):54–63. doi: 10.1093/jmedent/30.1.54. [DOI] [PubMed] [Google Scholar]
  30. Owen R. J. Chromosomal DNA fingerprinting--a new method of species and strain identification applicable to microbial pathogens. J Med Microbiol. 1989 Oct;30(2):89–99. doi: 10.1099/00222615-30-2-89. [DOI] [PubMed] [Google Scholar]
  31. Perng G. C., LeFebvre R. B., Johnson R. C. Further characterization of a potent immunogen and the chromosomal gene encoding it in the Lyme disease agent, Borrelia burgdorferi. Infect Immun. 1991 Jun;59(6):2070–2074. doi: 10.1128/iai.59.6.2070-2074.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. Preac-Mursic V., Wilske B., Schierz G. European Borrelia burgdorferi isolated from humans and ticks culture conditions and antibiotic susceptibility. Zentralbl Bakteriol Mikrobiol Hyg A. 1986 Dec;263(1-2):112–118. doi: 10.1016/s0176-6724(86)80110-9. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. Sigal L. H., Curran A. S. Lyme disease: a multifocal worldwide epidemic. Annu Rev Public Health. 1991;12:85–109. doi: 10.1146/annurev.pu.12.050191.000505. [DOI] [PubMed] [Google Scholar]
  36. 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]
  37. Stiernstedt G. Tick-borne Borrelia infection in Sweden. Scand J Infect Dis Suppl. 1985;45:1–70. doi: 10.3109/inf.1985.17.suppl-45.01. [DOI] [PubMed] [Google Scholar]
  38. Stålhammar-Carlemalm M., Jenny E., Gern L., Aeschlimann A., Meyer J. Plasmid analysis and restriction fragment length polymorphisms of chromosomal DNA allow a distinction between Borrelia burgdorferi strains. Zentralbl Bakteriol. 1990 Oct;274(1):28–39. doi: 10.1016/s0934-8840(11)80972-2. [DOI] [PubMed] [Google Scholar]
  39. Timms P., Eaves F. W., Girjes A. A., Lavin M. F. Comparison of Chlamydia psittaci isolates by restriction endonuclease and DNA probe analyses. Infect Immun. 1988 Jan;56(1):287–290. doi: 10.1128/iai.56.1.287-290.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wallich R., Helmes C., Schaible U. E., Lobet Y., Moter S. E., Kramer M. D., Simon M. M. Evaluation of genetic divergence among Borrelia burgdorferi isolates by use of OspA, fla, HSP60, and HSP70 gene probes. Infect Immun. 1992 Nov;60(11):4856–4866. doi: 10.1128/iai.60.11.4856-4866.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Welsh J., Pretzman C., Postic D., Saint Girons I., Baranton G., McClelland M. Genomic fingerprinting by arbitrarily primed polymerase chain reaction resolves Borrelia burgdorferi into three distinct phyletic groups. Int J Syst Bacteriol. 1992 Jul;42(3):370–377. doi: 10.1099/00207713-42-3-370. [DOI] [PubMed] [Google Scholar]
  42. 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]
  43. Zingg B. C., Brown R. N., Lane R. S., LeFebvre R. B. Genetic diversity among Borrelia burgdorferi isolates from wood rats and kangaroo rats in California. J Clin Microbiol. 1993 Dec;31(12):3109–3114. doi: 10.1128/jcm.31.12.3109-3114.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

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