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. 1995 Oct;33(10):2596–2600. doi: 10.1128/jcm.33.10.2596-2600.1995

Establishment of enzyme-linked immunosorbent assay using purified recombinant 83-kilodalton antigen of Borrelia burgdorferi sensu stricto and Borrelia afzelii for serodiagnosis of Lyme disease.

S Rauer 1, M Kayser 1, U Neubert 1, C Rasiah 1, A Vogt 1
PMCID: PMC228536  PMID: 8567889

Abstract

The 83-kDa antigen of Borrelia burgdorferi was expressed as a recombinant protein in Escherichia coli and purified for use in an enzyme-linked immunosorbent assay (p83-ELISA). Antibodies to the 83-kDa antigen of both the immunoglobulin G (IgG) and IgM isotypes could be detected in all stages of Lyme disease. Sensitivity varied, depending on the clinical stage of illness. In early stages, as defined for 118 patients with erythema migrans, it was found to be 20% (24 of 118 patients: 7 with IgM, 16 with IgG, and 1 with IgM and IgG). Of the patients with late-stage Lyme arthritis and acrodermatitis chronica atrophicans, 94% (16 of 17:2 with IgM and IgG and 14 with IgG) and 86% (36 of 42:2 with IgG and IgM and 34 with IgG) revealed positive results in the p83-ELISA, respectively. p83 displays sequence heterogeneity according to the genomospecies, but when the reactions of serum specimens from acrodermatitis chronica atrophicans patients and arthritis patients with p83 derived from representative strains of B. burgdorferi sensu stricto and Borrelia afzelii in ELISAs were compared, no differences in specificity and sensitivity were seen. When 82 serum specimens from healthy controls were tested, none had IgG and only 3 (4%) had IgM antibodies, indicating a high specificity. Positive reactions with antibodies against Treponema pallidum (1 of 37 patients; IgG) and Epstein-Barr virus (1 of 44 patients; IgM) and with autoantibodies of various specificities (1 of 53 patients; IgG) were seen with < 3% of the serum samples te11111111111111111111 high speficicity for B. burgdorferi.2+ 13% for IgM antibodies, the IgM p83-ELISA provided little diagnostic information for Lyme disease, whereas the IgG p83-ELISA appears to be a suita ;e test for serodiagnosis of advanced-stage Lyme disease.

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Selected References

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  1. Aguero-Rosenfeld M. E., Nowakowski J., McKenna D. F., Carbonaro C. A., Wormser G. P. Serodiagnosis in early Lyme disease. J Clin Microbiol. 1993 Dec;31(12):3090–3095. doi: 10.1128/jcm.31.12.3090-3095.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barbour A. G. Laboratory aspects of Lyme borreliosis. Clin Microbiol Rev. 1988 Oct;1(4):399–414. doi: 10.1128/cmr.1.4.399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bruckbauer H. R., Preac-Mursic V., Fuchs R., Wilske B. Cross-reactive proteins of Borrelia burgdorferi. Eur J Clin Microbiol Infect Dis. 1992 Mar;11(3):224–232. doi: 10.1007/BF02098084. [DOI] [PubMed] [Google Scholar]
  4. Coleman J. L., Benach J. L. Characterization of antigenic determinants of Borrelia burgdorferi shared by other bacteria. J Infect Dis. 1992 Apr;165(4):658–666. doi: 10.1093/infdis/165.4.658. [DOI] [PubMed] [Google Scholar]
  5. Ditton H. J., Neuss M., Zöller L. Evidence that Borrelia burgdorferi immunodominant proteins p100, p94 and p83 are identical. FEMS Microbiol Lett. 1992 Jul 15;73(3):217–220. doi: 10.1016/0378-1097(92)90633-y. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Dressler F., Whalen J. A., Reinhardt B. N., Steere A. C. Western blotting in the serodiagnosis of Lyme disease. J Infect Dis. 1993 Feb;167(2):392–400. doi: 10.1093/infdis/167.2.392. [DOI] [PubMed] [Google Scholar]
  8. Eiffert H., Schlott T., Hoppert M., Lotter H., Thomssen R. Identification of an endoflagellar associated protein in Borrelia burgdorferi. J Med Microbiol. 1992 Mar;36(3):209–214. doi: 10.1099/00222615-36-3-209. [DOI] [PubMed] [Google Scholar]
  9. Feder H. M., Jr, Gerber M. A., Luger S. W., Ryan R. W. False positive serologic tests for Lyme disease after varicella infection. N Engl J Med. 1991 Dec 26;325(26):1886–1887. doi: 10.1056/nejm199112263252615. [DOI] [PubMed] [Google Scholar]
  10. Fikrig E., Huguenel E. D., Berland R., Rahn D. W., Hardin J. A., Flavell R. A. Serologic diagnosis of Lyme disease using recombinant outer surface proteins A and B and flagellin. J Infect Dis. 1992 Jun;165(6):1127–1132. doi: 10.1093/infdis/165.6.1127. [DOI] [PubMed] [Google Scholar]
  11. Hansen K., Asbrink E. Serodiagnosis of erythema migrans and acrodermatitis chronica atrophicans by the Borrelia burgdorferi flagellum enzyme-linked immunosorbent assay. J Clin Microbiol. 1989 Mar;27(3):545–551. doi: 10.1128/jcm.27.3.545-551.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hansen K., Hindersson P., Pedersen N. S. Measurement of antibodies to the Borrelia burgdorferi flagellum improves serodiagnosis in Lyme disease. J Clin Microbiol. 1988 Feb;26(2):338–346. doi: 10.1128/jcm.26.2.338-346.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hansen K. Laboratory diagnostic methods in Lyme borreliosis. Clin Dermatol. 1993 Jul-Sep;11(3):407–414. doi: 10.1016/0738-081x(93)90097-v. [DOI] [PubMed] [Google Scholar]
  14. Hansen K., Pii K., Lebech A. M. Improved immunoglobulin M serodiagnosis in Lyme borreliosis by using a mu-capture enzyme-linked immunosorbent assay with biotinylated Borrelia burgdorferi flagella. J Clin Microbiol. 1991 Jan;29(1):166–173. doi: 10.1128/jcm.29.1.166-173.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jauris-Heipke S., Fuchs R., Hofmann A., Lottspeich F., Preac-Mursic V., Soutschek E., Will G., Wilske B. Molecular characterization of the p100 gene of Borrelia burgdorferi strain PKo. FEMS Microbiol Lett. 1993 Dec 1;114(2):235–241. doi: 10.1111/j.1574-6968.1993.tb06579.x. [DOI] [PubMed] [Google Scholar]
  16. Kaiser R. Variable CSF findings in early and late Lyme neuroborreliosis: a follow-up study in 47 patients. J Neurol. 1994 Dec;242(1):26–36. doi: 10.1007/BF00920571. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. 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]
  19. Luft B. J., Mudri S., Jiang W., Dattwyler R. J., Gorevic P. D., Fischer T., Munoz P., Dunn J. J., Schubach W. H. The 93-kilodalton protein of Borrelia burgdorferi: an immunodominant protoplasmic cylinder antigen. Infect Immun. 1992 Oct;60(10):4309–4321. doi: 10.1128/iai.60.10.4309-4321.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ma B., Christen B., Leung D., Vigo-Pelfrey C. Serodiagnosis of Lyme borreliosis by western immunoblot: reactivity of various significant antibodies against Borrelia burgdorferi. J Clin Microbiol. 1992 Feb;30(2):370–376. doi: 10.1128/jcm.30.2.370-376.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Magnarelli L. A., Anderson J. F., Barbour A. G. Enzyme-linked immunosorbent assays for Lyme disease: reactivity of subunits of Borrelia burgdorferi. J Infect Dis. 1989 Jan;159(1):43–49. doi: 10.1093/infdis/159.1.43. [DOI] [PubMed] [Google Scholar]
  22. Magnarelli L. A., Miller J. N., Anderson J. F., Riviere G. R. Cross-reactivity of nonspecific treponemal antibody in serologic tests for Lyme disease. J Clin Microbiol. 1990 Jun;28(6):1276–1279. doi: 10.1128/jcm.28.6.1276-1279.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Rasiah C., Rauer S., Gassmann G. S., Vogt A. Use of a hybrid protein consisting of the variable region of the Borrelia burgdorferi flagellin and part of the 83-kDa protein as antigen for serodiagnosis of Lyme disease. J Clin Microbiol. 1994 Apr;32(4):1011–1017. doi: 10.1128/jcm.32.4.1011-1017.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Robinson J. M., Pilot-Matias T. J., Pratt S. D., Patel C. B., Bevirt T. S., Hunt J. C. Analysis of the humoral response to the flagellin protein of Borrelia burgdorferi: cloning of regions capable of differentiating Lyme disease from syphilis. J Clin Microbiol. 1993 Mar;31(3):629–635. doi: 10.1128/jcm.31.3.629-635.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Stiernstedt G., Dattwyler R., Duray P. H., Hansen K., Jirous J., Johnson R. C., Karlsson M., Preac-Mursic V., Schwan T. G. Diagnostic tests in Lyme borreliosis. Scand J Infect Dis Suppl. 1991;77:136–142. [PubMed] [Google Scholar]
  27. Volkman D. J., Luft B. J., Gorevic P. D., Schultz J., Padovano L. Characterization of an immunoreactive 93-kDa core protein of Borrelia burgdorferi with a human IgG monoclonal antibody. J Immunol. 1991 May 1;146(9):3177–3182. [PubMed] [Google Scholar]
  28. Wilske B., Fingerle V., Herzer P., Hofmann A., Lehnert G., Peters H., Pfister H. W., Preac-Mursic V., Soutschek E., Weber K. Recombinant immunoblot in the serodiagnosis of Lyme borreliosis. Comparison with indirect immunofluorescence and enzyme-linked immunosorbent assay. Med Microbiol Immunol. 1993 Nov;182(5):255–270. doi: 10.1007/BF00579624. [DOI] [PubMed] [Google Scholar]
  29. Wilske B., Fingerle V., Preac-Mursic V., Jauris-Heipke S., Hofmann A., Loy H., Pfister H. W., Rössler D., Soutschek E. Immunoblot using recombinant antigens derived from different genospecies of Borrelia burgdorferi sensu lato. Med Microbiol Immunol. 1994 Feb;183(1):43–59. doi: 10.1007/BF00193630. [DOI] [PubMed] [Google Scholar]
  30. Zöller L., Burkard S., Schäfer H. Validity of western immunoblot band patterns in the serodiagnosis of Lyme borreliosis. J Clin Microbiol. 1991 Jan;29(1):174–182. doi: 10.1128/jcm.29.1.174-182.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Zöller L., Cremer J., Faulde M. Western blot as a tool in the diagnosis of Lyme borreliosis. Electrophoresis. 1993 Sep;14(9):937–944. doi: 10.1002/elps.11501401149. [DOI] [PubMed] [Google Scholar]

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