• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jcmPermissionsJournals.ASM.orgJournalJCM ArticleJournal InfoAuthorsReviewers
J Clin Microbiol. Feb 1995; 33(2): 419–427.
PMCID: PMC227960

Immunoblot interpretation criteria for serodiagnosis of early Lyme disease.

Abstract

We monitored the antibody responses of 55 treated patients with early Lyme disease and physician-documented erythema migrans. Six sequential serum samples were obtained from patients before, during, and until one year after antibiotic therapy and analyzed by in-house enzyme-linked immunosorbent (ELISA) and immunoblot assays. An immunoblot procedure utilizing a gradient gel and an image analysis system was developed. A relational database management system was used to analyze the results and provide criteria for early disease immunoblot interpretation. Recommended criteria for the immunoglobulin M (IgM) immunoblot are the recognition of two of three proteins (24, 39, and 41 kDa). The recommended criteria for a positive IgG immunoblot are the recognition of two of five proteins (20, 24 [> 19 intensity units], 35, 39, and 88 kDa). Alternatively, if band intensity cannot be measured, the 22-kDa protein can be substituted for the 24-kDa protein with only a small decrease in sensitivity. Monoclonal antibodies were used to identify all these proteins except the 35-kDa protein. With the proposed immunoblot interpretations, the sequential serum samples were examined. At visit 1, the day of diagnosis and initiation of treatment, 54.5% of the serum samples were either IgM or IgG positive. The peak antibody response, with 80% of the serum samples positive, occurred at visit 2, 8 to 12 days into treatment. The sensitivities of the IgM and IgG immunoblot for detecting patients that were seropositive into the study period were 58.5 and 54.6%, respectively, at visit 1 and 100% at visit 2. Twenty percent of the patients remained seronegative throughout the study. The specificities of the IgM and IgG immunoblots were 92 to 94% and 93 to 96%, respectively. The IgM immunoblot and ELISA were similar in sensitivities, whereas the IgG immunoblot had greater sensitivity than the IgG ELISA (P = 0.006).

Full Text

The Full Text of this article is available as a PDF (278K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Aguero-Rosenfeld ME, Nowakowski J, McKenna DF, Carbonaro CA, Wormser GP. Serodiagnosis in early Lyme disease. J Clin Microbiol. 1993 Dec;31(12):3090–3095. [PMC free article] [PubMed]
  • Barbour AG. Isolation and cultivation of Lyme disease spirochetes. Yale J Biol Med. 1984 Jul-Aug;57(4):521–525. [PMC free article] [PubMed]
  • Berger BW, Johnson RC, Kodner C, Coleman L. Cultivation of Borrelia burgdorferi from erythema migrans lesions and perilesional skin. J Clin Microbiol. 1992 Feb;30(2):359–361. [PMC free article] [PubMed]
  • Coleman JL, Benach JL. Isolation of antigenic components from the Lyme disease spirochete: their role in early diagnosis. J Infect Dis. 1987 Apr;155(4):756–765. [PubMed]
  • Craft JE, Fischer DK, Shimamoto GT, Steere AC. Antigens of Borrelia burgdorferi recognized during Lyme disease. Appearance of a new immunoglobulin M response and expansion of the immunoglobulin G response late in the illness. J Clin Invest. 1986 Oct;78(4):934–939. [PMC free article] [PubMed]
  • Dressler F, Whalen JA, Reinhardt BN, Steere AC. Western blotting in the serodiagnosis of Lyme disease. J Infect Dis. 1993 Feb;167(2):392–400. [PubMed]
  • Feder HM, Jr, Gerber MA, Luger SW, Ryan RW. Persistence of serum antibodies to Borrelia burgdorferi in patients treated for Lyme disease. Clin Infect Dis. 1992 Nov;15(5):788–793. [PubMed]
  • Fuchs R, Jauris S, Lottspeich F, Preac-Mursic V, Wilske B, Soutschek E. Molecular analysis and expression of a Borrelia burgdorferi gene encoding a 22 kDa protein (pC) in Escherichia coli. Mol Microbiol. 1992 Feb;6(4):503–509. [PubMed]
  • Fung BP, McHugh GL, Leong JM, Steere AC. Humoral immune response to outer surface protein C of Borrelia burgdorferi in Lyme disease: role of the immunoglobulin M response in the serodiagnosis of early infection. Infect Immun. 1994 Aug;62(8):3213–3221. [PMC free article] [PubMed]
  • Grodzicki RL, Steere AC. 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. [PubMed]
  • Hughes CA, Engstrom SM, Coleman LA, Kodner CB, Johnson RC. Protective immunity is induced by a Borrelia burgdorferi mutant that lacks OspA and OspB. Infect Immun. 1993 Dec;61(12):5115–5122. [PMC free article] [PubMed]
  • Karlsson M. Western immunoblot and flagellum enzyme-linked immunosorbent assay for serodiagnosis of Lyme borreliosis. J Clin Microbiol. 1990 Sep;28(9):2148–2150. [PMC free article] [PubMed]
  • Karlsson M, Möllegård I, Stiernstedt G, Wretlind B. Comparison of Western blot and enzyme-linked immunosorbent assay for diagnosis of Lyme borreliosis. Eur J Clin Microbiol Infect Dis. 1989 Oct;8(10):871–877. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • 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. [PMC free article] [PubMed]
  • Magnarelli LA, Miller JN, Anderson JF, Riviere GR. Cross-reactivity of nonspecific treponemal antibody in serologic tests for Lyme disease. J Clin Microbiol. 1990 Jun;28(6):1276–1279. [PMC free article] [PubMed]
  • Marconi RT, Samuels DS, Garon CF. Transcriptional analyses and mapping of the ospC gene in Lyme disease spirochetes. J Bacteriol. 1993 Feb;175(4):926–932. [PMC free article] [PubMed]
  • Mariash CN, Seelig S, Oppenheimer JH. A rapid, inexpensive, quantitative technique for the analysis of two-dimensional electrophoretograms. Anal Biochem. 1982 Apr;121(2):388–394. [PubMed]
  • Mayo DR, Vance DW., Jr Parvovirus B19 as the cause of a syndrome resembling Lyme arthritis in adults. N Engl J Med. 1991 Feb 7;324(6):419–420. [PubMed]
  • Nadal D, Taverna C, Hitzig WH. Immunoblot analysis of antibody binding to polypeptides of Borrelia burgdorferi in children with different clinical manifestations of Lyme disease. Pediatr Res. 1989 Oct;26(4):377–382. [PubMed]
  • Nadelman RB, Luger SW, Frank E, Wisniewski M, Collins JJ, Wormser GP. Comparison of cefuroxime axetil and doxycycline in the treatment of early Lyme disease. Ann Intern Med. 1992 Aug 15;117(4):273–280. [PubMed]
  • Padula SJ, Dias F, Sampieri A, Craven RB, Ryan RW. Use of recombinant OspC from Borrelia burgdorferi for serodiagnosis of early Lyme disease. J Clin Microbiol. 1994 Jul;32(7):1733–1738. [PMC free article] [PubMed]
  • Padula SJ, Sampieri A, Dias F, Szczepanski A, Ryan RW. Molecular characterization and expression of p23 (OspC) from a North American strain of Borrelia burgdorferi. Infect Immun. 1993 Dec;61(12):5097–5105. [PMC free article] [PubMed]
  • Rahn DW, Malawista SE. Lyme disease: recommendations for diagnosis and treatment. Ann Intern Med. 1991 Mar 15;114(6):472–481. [PubMed]
  • Rose CD, Fawcett PT, Singsen BH, Dubbs SB, Doughty RA. Use of western blot and enzyme-linked immunosorbent assays to assist in the diagnosis of Lyme disease. Pediatrics. 1991 Sep;88(3):465–470. [PubMed]
  • Sadziene A, Wilske B, Ferdows MS, Barbour AG. The cryptic ospC gene of Borrelia burgdorferi B31 is located on a circular plasmid. Infect Immun. 1993 May;61(5):2192–2195. [PMC free article] [PubMed]
  • Saltzman RL, Quirk MR, Mariash CN, Jordan MC. Quantitation of cytomegalovirus DNA by blot hybridization in blood leukocytes of viremic patients. J Virol Methods. 1990 Oct;30(1):67–77. [PubMed]
  • Shrestha M, Grodzicki RL, Steere AC. Diagnosing early Lyme disease. Am J Med. 1985 Feb;78(2):235–240. [PubMed]
  • Simpson WJ, Schrumpf ME, Schwan TG. Reactivity of human Lyme borreliosis sera with a 39-kilodalton antigen specific to Borrelia burgdorferi. J Clin Microbiol. 1990 Jun;28(6):1329–1337. [PMC free article] [PubMed]
  • Steere AC, Grodzicki RL, Kornblatt AN, Craft JE, Barbour AG, Burgdorfer W, Schmid GP, Johnson E, Malawista SE. The spirochetal etiology of Lyme disease. N Engl J Med. 1983 Mar 31;308(13):733–740. [PubMed]
  • Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. [PMC free article] [PubMed]
  • 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. [PMC free article] [PubMed]
  • Wilske B, Preac-Mursic V, Schierz G, Busch KV. Immunochemical and immunological analysis of European Borrelia burgdorferi strains. Zentralbl Bakteriol Mikrobiol Hyg A. 1986 Dec;263(1-2):92–102. [PubMed]
  • 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. [PMC free article] [PubMed]

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

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Cited in Books
    Cited in Books
    PubMed Central articles cited in books
  • MedGen
    MedGen
    Related information in MedGen
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...