Skip to main content
NCBI home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1996 Oct;34(10):2343–2350. doi: 10.1128/jcm.34.10.2343-2350.1996

New laboratory guidelines for serologic diagnosis of Lyme disease: evaluation of the two-test protocol.

T B Ledue 1, M F Collins 1, W Y Craig 1
PMCID: PMC229265  PMID: 8880477

Abstract

Recent guidelines established by the Association of State and Territorial Public Health Laboratory Directors (ASTPHLD) and the U.S. Centers for Disease Control and Prevention (CDC) recommend the use of a two-test protocol for the serologic diagnosis of Lyme disease (LD). The two-test protocol relies on a sensitive screening test, which is followed by specific immunoglobulin M (IgM) and/or IgG immunoblotting (IB), depending on the date of disease onset, of all samples with equivocal and positive screening test results. We evaluated a commercially available IgM-IgG enzyme-linked immunosorbent assay (ELISA) and separate IB tests for IgM and IgG antibodies to Borrelia burgdorferi as candidate assays for the two-test protocol. Serum samples obtained from healthy controls (n = 29), from patients with diagnoses or laboratory findings associated with serologic cross-reactivity to LD (n = 24), and from patients with well-documented early- and late-stage LD provided by the CDC and the College of American Pathologists (n = 53) were examined to determine each assay's individual sensitivity and specificity. No false-positive results were detected among the healthy controls by either ELISA or IB, whereas four false-positive ELISA results were recorded within the cross-reactive group. None of these sera, however, were positive for either IgM or IgG reactivity according to IB band criteria. With regard to the patients with LD, we determined the sensitivity and specificity of the ELISA to be 96 and 100%, respectively, compared with the reference data provided for these specimens. When we compared our IB results with data from CDC, the assay sensitivity and specificity were 80 and 96.2%, respectively, for IgM and 81.8 and 95.8%, respectively, for IgG. Pursuant to this evaluation we assessed the suitability of the two-test protocol by performing a retrospective analysis using clinical history to define samples as positive or negative for LD. We determined clinical sensitivity and specificity for all study subjects (n = 112) to be 50 and 100%, respectively. A reduction in the clinical sensitivity of the two-test protocol was associated with a lack of antibody response or seroconversion in LD patients treated with antibiotics. We conclude that the CDC-ASTPHLD guidelines provide useful criteria for test performance and interpretation aimed at standardizing the serologic diagnosis of LD.

Full Text

The Full Text of this article is available as a PDF (241.8 KB).

Selected References

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

  1. Aguero-Rosenfeld M. E., Nowakowski J., Bittker S., Cooper D., Nadelman R. B., Wormser G. P. Evolution of the serologic response to Borrelia burgdorferi in treated patients with culture-confirmed erythema migrans. J Clin Microbiol. 1996 Jan;34(1):1–9. doi: 10.1128/jcm.34.1.1-9.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Bakken L. L., Case K. L., Callister S. M., Bourdeau N. J., Schell R. F. Performance of 45 laboratories participating in a proficiency testing program for Lyme disease serology. JAMA. 1992 Aug 19;268(7):891–895. [PubMed] [Google Scholar]
  4. 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]
  5. Cutler S. J., Wright D. J., Luckhurst V. H. Simplified method for the interpretation of immunoblots for Lyme borreliosis. FEMS Immunol Med Microbiol. 1993 Apr;6(4):281–285. doi: 10.1111/j.1574-695X.1993.tb00340.x. [DOI] [PubMed] [Google Scholar]
  6. Dattwyler R. J., Volkman D. J., Luft B. J., Halperin J. J., Thomas J., Golightly M. G. Seronegative Lyme disease. Dissociation of specific T- and B-lymphocyte responses to Borrelia burgdorferi. N Engl J Med. 1988 Dec 1;319(22):1441–1446. doi: 10.1056/NEJM198812013192203. [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. Engstrom S. M., Shoop E., Johnson R. C. Immunoblot interpretation criteria for serodiagnosis of early Lyme disease. J Clin Microbiol. 1995 Feb;33(2):419–427. doi: 10.1128/jcm.33.2.419-427.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fawcett P. T., Gibney K. M., Rose C. D., Dubbs S. B., Doughty R. A. Frequency and specificity of antibodies that crossreact with Borrelia burgdorferi antigens. J Rheumatol. 1992 Apr;19(4):582–587. [PubMed] [Google Scholar]
  10. Golightly M. G. Laboratory considerations in the diagnosis and management of Lyme borreliosis. Am J Clin Pathol. 1993 Feb;99(2):168–174. doi: 10.1093/ajcp/99.2.168. [DOI] [PubMed] [Google Scholar]
  11. Grodzicki R. L., Steere A. C. Comparison of immunoblotting and indirect enzyme-linked immunosorbent assay using different antigen preparations for diagnosing early Lyme disease. J Infect Dis. 1988 Apr;157(4):790–797. doi: 10.1093/infdis/157.4.790. [DOI] [PubMed] [Google Scholar]
  12. Hilton E., Devoti J., Sood S. Recommendation to include OspA and OspB in the new immunoblotting criteria for serodiagnosis of Lyme disease. J Clin Microbiol. 1996 Jun;34(6):1353–1354. doi: 10.1128/jcm.34.6.1353-1354.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Johnson D. E., Weinberg L., Ciarkowski J., West P., Colwell R. R. Wound infection caused by Kanagawa-negative Vibrio parahaemolyticus. J Clin Microbiol. 1984 Oct;20(4):811–812. doi: 10.1128/jcm.20.4.811-812.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kowal K., Weinstein A. Western blot band intensity analysis. Application to the diagnosis of Lyme arthritis. Arthritis Rheum. 1994 Aug;37(8):1206–1211. doi: 10.1002/art.1780370815. [DOI] [PubMed] [Google Scholar]
  15. Lovece S., Stern R., Kagen L. J. Effects of rheumatoid factor, antinuclear antibodies and plasma reagin on the serologic assay for Lyme disease. J Rheumatol. 1991 Dec;18(12):1813–1818. [PubMed] [Google Scholar]
  16. Magnarelli L. A. Serologic diagnosis of Lyme disease. Ann N Y Acad Sci. 1988;539:154–161. doi: 10.1111/j.1749-6632.1988.tb31848.x. [DOI] [PubMed] [Google Scholar]
  17. Padula S. J., Sampieri A., Dias F., Szczepanski A., Ryan R. W. Molecular characterization and expression of p23 (OspC) from a North American strain of Borrelia burgdorferi. Infect Immun. 1993 Dec;61(12):5097–5105. doi: 10.1128/iai.61.12.5097-5105.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rahn D. W., Malawista S. E. Lyme disease: recommendations for diagnosis and treatment. Ann Intern Med. 1991 Mar 15;114(6):472–481. doi: 10.7326/0003-4819-114-6-472. [DOI] [PubMed] [Google Scholar]
  19. Schmitz J. L., Powell C. S., Folds J. D. Comparison of seven commercial kits for detection of antibodies to Borrelia burgdorferi. Eur J Clin Microbiol Infect Dis. 1993 Jun;12(6):419–424. doi: 10.1007/BF01967435. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Schwan T. G., Simpson W. J. Factors influencing the antigenic reactivity of Borrelia burgdorferi, the Lyme disease spirochete. Scand J Infect Dis Suppl. 1991;77:94–101. [PubMed] [Google Scholar]
  22. Shrestha M., Grodzicki R. L., Steere A. C. Diagnosing early Lyme disease. Am J Med. 1985 Feb;78(2):235–240. doi: 10.1016/0002-9343(85)90432-2. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Steere A. C. Lyme disease. N Engl J Med. 1989 Aug 31;321(9):586–596. doi: 10.1056/NEJM198908313210906. [DOI] [PubMed] [Google Scholar]
  25. Steere A. C., Taylor E., McHugh G. L., Logigian E. L. The overdiagnosis of Lyme disease. JAMA. 1993 Apr 14;269(14):1812–1816. [PubMed] [Google Scholar]
  26. Wilske B., Jauris-Heipke S., Lobentanzer R., Pradel I., Preac-Mursic V., Rössler D., Soutschek E., Johnson R. C. Phenotypic analysis of outer surface protein C (OspC) of Borrelia burgdorferi sensu lato by monoclonal antibodies: relationship to genospecies and OspA serotype. J Clin Microbiol. 1995 Jan;33(1):103–109. doi: 10.1128/jcm.33.1.103-109.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES