This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Engstrom, S. M. Articles by Johnson, R. C. Search for Related Content PubMed PubMed Citation Articles by Engstrom, S. M. Articles by Johnson, R. C.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, 02 1995, 419-427, Vol 33, No. 2
Copyright © 1995 by the American Society for Microbiology. All rights reserved.

Immunoblot interpretation criteria for serodiagnosis of early Lyme disease

SM Engstrom, E Shoop and RC Johnson
Department of Microbiology, University of Minnesota Medical School, Minneapolis.

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).

This article has been cited by other articles:

• Barbour, A. G., Jasinskas, A., Kayala, M. A., Davies, D. H., Steere, A. C., Baldi, P., Felgner, P. L. (2008). A Genome-Wide Proteome Array Reveals a Limited Set of Immunogens in Natural Infections of Humans and White-Footed Mice with Borrelia burgdorferi. Infect. Immun. 76: 3374-3389 [Abstract] [Full Text]  
• Hsieh, Y.-F., Liu, H.-W., Hsu, T.-C., Wei, J. C.-C., Shih, C.-M., Krause, P. J., Tsay, G. J. (2007). Serum Reactivity against Borrelia burgdorferi OspA in Patients with Rheumatoid Arthritis. CVI 14: 1437-1441 [Abstract] [Full Text]  
• Vojdani, A., Hebroni, F., Raphael, Y., Erde, J., Raxlen, B. (2007). Novel Diagnosis of Lyme Disease: Potential for CAM Intervention. Evid Based Complement Alternat Med 0: nem138v1-nem138 [Abstract] [Full Text]  
• Croda, J., Ramos, J. G. R., Matsunaga, J., Queiroz, A., Homma, A., Riley, L. W., Haake, D. A., Reis, M. G., Ko, A. I. (2007). Leptospira Immunoglobulin-Like Proteins as a Serodiagnostic Marker for Acute Leptospirosis. J. Clin. Microbiol. 45: 1528-1534 [Abstract] [Full Text]  
• Wormser, G. P. (2006). Clinical practice. Early Lyme disease.. NEJM 354: 2794-2801 [Full Text]  
• Evans, R., Mavin, S., Ho-Yen, D. O (2005). Audit of the laboratory diagnosis of Lyme disease in Scotland. J Med Microbiol 54: 1139-1141 [Abstract] [Full Text]  
• Philipp, M. T., Wormser, G. P., Marques, A. R., Bittker, S., Martin, D. S., Nowakowski, J., Dally, L. G. (2005). A Decline in C6 Antibody Titer Occurs in Successfully Treated Patients with Culture-Confirmed Early Localized or Early Disseminated Lyme Borreliosis. CVI 12: 1069-1074 [Abstract] [Full Text]  
• Aguero-Rosenfeld, M. E., Wang, G., Schwartz, I., Wormser, G. P. (2005). Diagnosis of Lyme Borreliosis. Clin. Microbiol. Rev. 18: 484-509 [Abstract] [Full Text]  
• Ramamoorthy, R., McClain, N. A., Gautam, A., Scholl-Meeker, D. (2005). Expression of the bmpB Gene of Borrelia burgdorferi Is Modulated by Two Distinct Transcription Termination Events. J. Bacteriol. 187: 2592-2600 [Abstract] [Full Text]  
• Brisson, D., Dykhuizen, D. E. (2004). ospC Diversity in Borrelia burgdorferi: Different Hosts Are Different Niches. Genetics 168: 713-722 [Abstract] [Full Text]  
• Miller, J. C., von Lackum, K., Babb, K., McAlister, J. D., Stevenson, B. (2003). Temporal Analysis of Borrelia burgdorferi Erp Protein Expression throughout the Mammal-Tick Infectious Cycle. Infect. Immun. 71: 6943-6952 [Abstract] [Full Text]  
• Philipp, M. T., Marques, A. R., Fawcett, P. T., Dally, L. G., Martin, D. S. (2003). C6 Test as an Indicator of Therapy Outcome for Patients with Localized or Disseminated Lyme Borreliosis. J. Clin. Microbiol. 41: 4955-4960 [Abstract] [Full Text]  
• Priya, C. G., Bhavani, K., Rathinam, S. R., Muthukkaruppan, V. R. (2003). Identification and evaluation of LPS antigen for serodiagnosis of uveitis associated with leptospirosis. J Med Microbiol 52: 667-673 [Abstract] [Full Text]  
• Hefty, P. S., Brooks, C. S., Jett, A. M., White, G. L., Wikel, S. K., Kennedy, R. C., Akins, D. R. (2002). OspE-Related, OspF-Related, and Elp Lipoproteins Are Immunogenic in Baboons Experimentally Infected with Borrelia burgdorferi and in Human Lyme Disease Patients. J. Clin. Microbiol. 40: 4256-4265 [Abstract] [Full Text]  
• PANELIUS, J., LAHDENNE, P., HEIKKILA, T., PELTOMAA, M., OKSI, J., SEPPALA, I. (2002). Recombinant OspC from Borrelia burgdorferi sensu stricto, B. afzelii and B. garinii in the serodiagnosis of Lyme borreliosis. J Med Microbiol 51: 731-739 [Abstract] [Full Text]  
• Steere, A. C. (2002). A 58-Year-Old Man With a Diagnosis of Chronic Lyme Disease. JAMA 288: 1002-1010 [Full Text]  
• Marques, A. R., Martin, D. S., Philipp, M. T. (2002). Evaluation of the C6 Peptide Enzyme-Linked Immunosorbent Assay for Individuals Vaccinated with the Recombinant OspA Vaccine. J. Clin. Microbiol. 40: 2591-2593 [Abstract] [Full Text]  
• Heikkila, T., Seppala, I., Saxen, H., Panelius, J., Peltomaa, M., Julin, T., Carlsson, S.-A., Lahdenne, P. (2002). Recombinant BBK32 Protein in Serodiagnosis of Early and Late Lyme Borreliosis. J. Clin. Microbiol. 40: 1174-1180 [Abstract] [Full Text]  
• Smith, R. P., Schoen, R. T., Rahn, D. W., Sikand, V. K., Nowakowski, J., Parenti, D. L., Holman, M. S., Persing, D. H., Steere, A. C. (2002). Clinical Characteristics and Treatment Outcome of Early Lyme Disease in Patients with Microbiologically Confirmed Erythema Migrans. ANN INTERN MED 136: 421-428 [Abstract] [Full Text]  
• Vaz, A., Glickstein, L., Field, J. A., McHugh, G., Sikand, V. K., Damle, N., Steere, A. C. (2001). Cellular and Humoral Immune Responses to Borrelia burgdorferi Antigens in Patients with Culture-Positive Early Lyme Disease. Infect. Immun. 69: 7437-7444 [Abstract] [Full Text]  
• Brunner, M., Sigal, L. H. (2001). Use of Serum Immune Complexes in a New Test That Accurately Confirms Early Lyme Disease and Active Infection with Borrelia burgdorferi. J. Clin. Microbiol. 39: 3213-3221 [Abstract] [Full Text]  
• Flannery, B., Costa, D., Carvalho, F. P., Guerreiro, H., Matsunaga, J., Da Silva, E. D., Ferreira, A. G. P., Riley, L. W., Reis, M. G., Haake, D. A., Ko, A. I. (2001). Evaluation of Recombinant Leptospira Antigen-Based Enzyme-Linked Immunosorbent Assays for the Serodiagnosis of Leptospirosis. J. Clin. Microbiol. 39: 3303-3310 [Abstract] [Full Text]  
• Steere, A. C. (2001). Lyme Disease. NEJM 345: 115-125 [Full Text]  
• Carroll, J. A., Cordova, R. M., Garon, C. F. (2000). Identification of 11 pH-Regulated Genes in Borrelia burgdorferi Localizing to Linear Plasmids. Infect. Immun. 68: 6677-6684 [Abstract] [Full Text]  
• Munson, E. L., Du Chateau, B. K., Jobe, D. A., Lovrich, S. D., Callister, S. M., Schell, R. F. (2000). Production of Borreliacidal Antibody to Outer Surface Protein A In Vitro and Modulation by Interleukin-4. Infect. Immun. 68: 5496-5501 [Abstract] [Full Text]  
• Hovius, J. W. R., Hovius, K. E., Oei, A., Houwers, D. J., van Dam, A. P. (2000). Antibodies against Specific Proteins of and Immobilizing Activity against Three Strains of Borrelia burgdorferi Sensu Lato Can Be Found in Symptomatic but Not in Infected Asymptomatic Dogs. J. Clin. Microbiol. 38: 2611-2621 [Abstract] [Full Text]  
• Robertson, J., Guy, E., Andrews, N., Wilske, B., Anda, P., Granström, M., Hauser, U., Moosmann, Y., Sambri, V., Schellekens, J., Stanek, G., Gray, J. (2000). A European Multicenter Study of Immunoblotting in Serodiagnosis of Lyme Borreliosis. J. Clin. Microbiol. 38: 2097-2102 [Abstract] [Full Text]  
• Magnarelli, L. A., Ijdo, J. W., Padula, S. J., Flavell, R. A., Fikrig, E. (2000). Serologic Diagnosis of Lyme Borreliosis by Using Enzyme-Linked Immunosorbent Assays with Recombinant Antigens. J. Clin. Microbiol. 38: 1735-1739 [Abstract] [Full Text]  
• Miller, J. C., El-Hage, N., Babb, K., Stevenson, B. (2000). Borrelia burgdorferi B31 Erp Proteins That Are Dominant Immunoblot Antigens of Animals Infected with Isolate B31 Are Recognized by Only a Subset of Human Lyme Disease Patient Sera. J. Clin. Microbiol. 38: 1569-1574 [Abstract] [Full Text]  
• Wieneke, C. A., Lovrich, S. D., Callister, S. M., Jobe, D. A., Marks, J. A., Schell, R. F. (2000). Evaluation of Whole-Cell and OspC Enzyme-Linked Immunosorbent Assays for Discrimination of Early Lyme Borreliosis from OspA Vaccination. J. Clin. Microbiol. 38: 313-317 [Abstract] [Full Text]  
• Liang, F. T., Steere, A. C., Marques, A. R., Johnson, B. J. B., Miller, J. N., Philipp, M. T. (1999). Sensitive and Specific Serodiagnosis of Lyme Disease by Enzyme-Linked Immunosorbent Assay with a Peptide Based on an Immunodominant Conserved Region of Borrelia burgdorferi VlsE. J. Clin. Microbiol. 37: 3990-3996 [Abstract] [Full Text]  
• Ryffel, K., Péter, O., Rutti, B., Suard, A., Dayer, E. (1999). Scored Antibody Reactivity Determined by Immunoblotting Shows an Association between Clinical Manifestations and Presence of Borrelia burgdorferi sensu stricto, B. garinii, B. afzelii, and B. Valaisiana in Humans. J. Clin. Microbiol. 37: 4086-4092 [Abstract] [Full Text]  
• Aguero-Rosenfeld, M. E., Roberge, J., Carbonaro, C. A., Nowakowski, J., Nadelman, R. B., Wormser, G. P. (1999). Effects of OspA Vaccination on Lyme Disease Serologic Testing. J. Clin. Microbiol. 37: 3718-3721 [Abstract] [Full Text]  
• Blaauw, A. A. M., van Loon, A. M., Schellekens, J. F. P., Bijlsma, J. W. J. (1999). Clinical evaluation of guidelines and two-test approach for Lyme disease. Rheumatology (Oxford) 38: 1121-1126 [Abstract] [Full Text]  
• Hauser, U., Lehnert, G., Wilske, B. (1999). Validity of Interpretation Criteria for Standardized Western Blots (Immunoblots) for Serodiagnosis of Lyme Borreliosis Based on Sera Collected throughout Europe. J. Clin. Microbiol. 37: 2241-2247 [Abstract] [Full Text]  
• Gilmore, R. D. Jr., Murphree, R. L., James, A. M., Sullivan, S. A., Johnson, B. J. B. (1999). The Borrelia burgdorferi 37-Kilodalton Immunoblot Band (P37) Used in Serodiagnosis of Early Lyme Disease Is the flaA Gene Product. J. Clin. Microbiol. 37: 548-552 [Abstract] [Full Text]  
• Wang, I.-N., Dykhuizen, D. E., Qiu, W., Dunn, J. J., Bosler, E. M., Luft, B. J. (1999). Genetic Diversity of ospC in a Local Population of Borrelia burgdorferi sensu stricto. Genetics 151: 15-30 [Abstract] [Full Text]  
• Stevenson, B., Bono, J. L., Elias, A., Tilly, K., Rosa, P. (1998). Transformation of the Lyme Disease Spirochete Borrelia burgdorferi with Heterologous DNA. J. Bacteriol. 180: 4850-4855 [Abstract] [Full Text]  
• Brettschneider, S., Bruckbauer, H., Klugbauer, N., Hofmann, H. (1998). Diagnostic Value of PCR for Detection of Borrelia burgdorferi in Skin Biopsy and Urine Samples from Patients with Skin Borreliosis. J. Clin. Microbiol. 36: 2658-2665 [Abstract] [Full Text]  
• Hauser, U., Lehnert, G., Wilske, B. (1998). Diagnostic Value of Proteins of Three Borrelia Species (Borrelia burgdorferi Sensu Lato) and Implications for Development and Use of Recombinant Antigens for Serodiagnosis of Lyme Borreliosis in Europe. CVI 5: 456-462 [Abstract] [Full Text]  
• Fawcett, P. T., Rosé, C. D., Gibney, K. M., Doughty, R. A. (1998). Comparison of Immunodot and Western Blot Assays for Diagnosing Lyme Borreliosis. CVI 5: 503-506 [Abstract] [Full Text]  
• Rauer, S., Spohn, N., Rasiah, C., Neubert, U., Vogt, A. (1998). Enzyme-Linked Immunosorbent Assay Using Recombinant OspC and the Internal 14-kDa Flagellin Fragment for Serodiagnosis of Early Lyme Disease. J. Clin. Microbiol. 36: 857-861 [Abstract] [Full Text]  
• Brunner, M., Stein, S., Mitchell, P. D., Sigal, L. H. (1998). Immunoglobulin M Capture Assay for Serologic Confirmation of Early Lyme Disease: Analysis of Immune Complexes with Biotinylated Borrelia burgdorferi Sonicate Enhanced with Flagellin Peptide Epitope. J. Clin. Microbiol. 36: 1074-1080 [Abstract] [Full Text]  
• Hauser, U., Krahl, H., Peters, H., Fingerle, V., Wilske, B. (1998). Impact of Strain Heterogeneity on Lyme Disease Serology in Europe: Comparison of Enzyme-Linked Immunosorbent Assays Using Different Species of Borrelia burgdorferi Sensu Lato. J. Clin. Microbiol. 36: 427-436 [Abstract] [Full Text]  
• Priem, S., Burmester, G. R, Kamradt, T., Wolbart, K., Rittig, M. G, Krause, A. (1998). Detection of Borrelia burgdorferi by polymerase chain reaction in synovial membrane, but not in synovial fluid from patients with persisting Lyme arthritis after antibiotic therapy. Ann Rheum Dis 57: 118-121 [Abstract] [Full Text]