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Journal of Clinical Microbiology, November 2001, p. 4013-4019, Vol. 39, No. 11
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.11.4013-4019.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Recombinant Flagellin A Proteins from Borrelia burgdorferi Sensu Stricto, B. afzelii, and B. garinii in Serodiagnosis of Lyme Borreliosis

Jaana Panelius,1,* Pekka Lahdenne,2 Harri Saxen,2 Tero Heikkilä,1,2 and Ilkka Seppälä1,3

Haartman Institute, Department of Bacteriology and Immunology,1 and Hospital for Children and Adolescents,2 University of Helsinki, and HUCH Laboratory Diagnostics, Helsinki University Central Hospital,3 Helsinki, Finland

Received 19 March 2001/Returned for modification 1 July 2001/Accepted 3 September 2001

Genes for flagellin A (FlaA) proteins from European borrelial strains of Borrelia burgdorferi sensu stricto, B. afzelii, and B. garinii were cloned and sequenced. An identity of 92 to 93% was observed in the flaA sequences of the different species. Polyhistidine-tagged recombinant FlaA (rFlaA) proteins were produced in Escherichia coli and used as antigens in Western blotting (WB) and enzyme-linked immunosorbent assay (ELISA). In immunoglobulin G (IgG) WB, 71% (10 of 14) of the sera from neuroborreliosis and 86% (12 of 14) of those from Lyme arthritis patients reacted with one to three rFlaAs. In IgG ELISA, 74% (14 of 19) and 79% (15 of 19) of patients with neuroborreliosis and arthritis, respectively, were positive. The immunoreactivity in local European patient sera was stronger against rFlaA from B. garinii and B. afzelii than against rFlaA from B. burgdorferi sensu stricto. Neither IgG nor IgM ELISA was sensitive in the serodiagnosis of erythema migrans. Serum samples from patients with syphilis and systemic lupus erythematosus showed mild cross-reactivity in IgG tests. Sera from Yersinia enterocolitica or beta-hemolytic Streptococcus infections showed only occasional responses. With IgM ELISA, 58% (11 of 19) and 37% (7 of 19) of patients with neuroborreliosis and arthritis, respectively, were positive. Cross-reactive antibodies to FlaA, especially in serum samples from patients with rheumatoid factor positivity and Epstein-Barr virus infection, reduced the specificity of IgM serodiagnosis. Therefore, rFlaA seems to have a limited role for IgM serodiagnosis, yet rFlaA might be useful in the IgG serodiagnosis of disseminated Lyme borreliosis.

* Corresponding author. Mailing address: Haartman Institute, Department of Bacteriology and Immunology, P.O. Box 21, University of Helsinki, Helsinki, FIN-00014, Finland. Phone: (358)-9-1911. Fax: (358)-9-19126382. E-mail: jaana.panelius{at}helsinki.fi.

Journal of Clinical Microbiology, November 2001, p. 4013-4019, Vol. 39, No. 11
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.11.4013-4019.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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