Skip to main content

Borrelia burgdorferi DNA in the urine of treated patients with chronic lyme disease symptoms. A PCR study of 97 cases

Borrelia burgdorferi DNA im Urin von Patienten mit chronischer Lyme-Krankheit nach Therapie. PCR-Studie mit 97 Fällen

Summary

The presence ofBorrelia burgdorferi DNA was established by PCR from urine samples of 97 patients clinically diagnosed as presenting with symptoms of chronic Lyme disease. All patients had shown erythema chronica migrans following a deer tick bite. Most of the patients had been antibiotic-treated for extended periods of time. We used three sets of primer pairs with DNA sequences for the gene coding of outer surface protein A (OspA) and of a genomic sequence ofB. burgdorferi to study samples of physician-referred patients from the mideastern USA. Controls from 62 healthy volunteers of the same geographic areas were routinely carried through the procedures in parallel with patients' samples. Of the 97 patients, 72 (74.2%) were found with positive PCR and the rest with negative PCR. The 62 healthy volunteers were PCR negative. It is proposed that a sizeable group of patients diagnosed on clinical grounds as having chronic Lyme disease may still excreteBorrelia DNA, and may do so in spite of intensive antibiotic treatment.

Zusammenfassung

Bei 97 Patienten mit klinisch verifizierter chronischer Lyme-Krankheit wurde mittels PCR in Urinproben nachBorrelia burgdorferi-DNA gesucht. Bei allen Patienten war nach Zeckenstich ein Erythema chronicum migrans aufgetreten. Bei den meisten Patienten war eine langfristige Antibiotikabehandlung erfolgt. Wir verwendeten drei Primerpaare mit DNA-Sequenzen für das Gen, das für das Oberflächenprotein A (OSP A) kodiert und für eine genomische Sequenz vonB. burgdorferi, um Proben von Patienten zu untersuchen, die von Ärzten aus dem mittleren Osten der USA überwiesen worden waren. Proben von 62 gesunden Freiwilligen derselben geographischen Regionen wurden routinemäßig parallel zu den Patientenproben getestet. 72 der 92 Patienten (74,2%) hatten ein positives Testergebnis, die übrigen waren negativ. Auch die 62 Probanden waren PCR-negativ. Ein beträchtlicher Anteil der Patienten mit klinisch diagnostizierter Krankheit scheidet offensichtlichBorrelia DNA aus, auch wenn eine intensive Antibiotikatherapie vorausgegangen ist.

References

  1. 1.

    Steere, A. C. Borrelia burgdorferi (Lyme disease, Lyme borreliosis). In:Mandell, G. L., Bennett, J. E., Dolin, R. (eds): Mandell, Douglas and Bennett's principles and practice of infectious diseases. Churchill Livingstone, New York 1995, pp. 2143–2155.

    Google Scholar 

  2. 2.

    Rahn, D. W., Malawista, S. E. Lyme disease: recommendations for diagnosis and treatment. Ann. Intern. Med. 114 (1991) 472–481.

    Google Scholar 

  3. 3.

    Bakken, L. L., Case, K. L., Callister, S. M., Bordeau, N. J., Schell, R. F. Performance of 45 laboratories participating in a proficiency testing program for Lyme disease serology. JAMA 268 (1992) 891–895.

    Google Scholar 

  4. 4.

    Aguero-Rosenfeld, M. E., Nowakoswki, J., Bittker, S., Cooper, D., Nadelman, R. B., Wormser, G. P. Evolution of the serologic response toBorrelia burgdorferi in treated patients with culture-confirmed erythema migrans. J. Clin. Microbiol. 34 (1996) 1–9.

    Google Scholar 

  5. 5.

    Dressler, F., Whalen, J. A., Reinhardt, B. N., Steere, A. Western blotting in the serodiagnosis of Lyme disease. J. Infect. Dis. 167 (1993) 392–400.

    Google Scholar 

  6. 6.

    Engstrom, S. M., Shoop, E., Johnson, R. C. Immunoblot interpretation criteria for serodiagnosis of early Lyme disease. J. Clin. Microbiol. 33 (1995) 419–427.

    Google Scholar 

  7. 7.

    Padula, S. J., Samperi, A., Dias, F., Szczepanski, A., Ryan, R. W. Molecular characterization and expression p p23 (OspC) from a North American strain ofB. burgdorferi. Infect. Immun. 61 (1993) 5097–5105.

    Google Scholar 

  8. 8.

    Magnarelli, L. A., Fikrig, E., Berland, R., Anderson, J. F., Flavell, R. A. Comparison of whole-cell antibodies and an antigenic flagellar epitope ofBorrelia burgdorferi in serologic tests for diagnosis of Lyme borreliosis. J. Clin. Microbiol. 30 (1992) 3158–3162.

    Google Scholar 

  9. 9.

    Oksi, J., Uksila, J., Marjamäki, M., Nikoskelainen, J., Viljanen, M. K. Antibodies against whole sonicatedBorrelia burgdorferi spirochetes, 41-kilodalton flagellin, and P39 protein in patients with PCR- or culture-proven late Lyme borreliosis. J. Clin. Microbiol. 33 (1995) 2260–2264.

    Google Scholar 

  10. 10.

    Berger, B. W., Kaplan, M. H., Rothenberg, I. R., Barbour, A. G. Isolation and characterization of the Lyme disease spirochete from the skin of patients with erythema chronicum migrans. J. Am. Acad. Dermatol. 13 (1985) 444–449.

    Google Scholar 

  11. 11.

    Benach, J. L., Bosler, E. M., Hanrahan, J. P., Coleman, J. L., Habicht, G. S., Bast, T. F., Cameron, D. J., Ziegler, J. L., Barbour, A. G., Burgdorfer, W., Edelman, R., Kaslow, R. A. Spirochetes isolated from the blood of two patients with Lyme disease. N. Engl. J. Med. 308 (1983) 740–742.

    Google Scholar 

  12. 12.

    Wallach, F. R., Forni, A. L., Hariprashad, J., Stoeckle, M. Y., Steinberg, C. R., Fisher, L., Malawista, S. E., Murray, H. W. CirculatingBorrelia burgdorferi in patients with acute Lyme disease: results of blood cultures and serum DNA analysis. J. Infect. Dis. 168 (1993) 1541–1543.

    Google Scholar 

  13. 13.

    Nocton, J. J., Dressler, F., Rutledge, B. J., Rys, P. N., Persing, D. H., Steere, A. C. Detection ofBorrelia burgdorferi DNA by polymerase chain reaction in synovial fluid from patients with Lyme arthritis. N. Engl. J. Med. 330 (1994) 229–234.

    Google Scholar 

  14. 14.

    Malawista, S. E., Barthold, S. W., Persing, D. Fate ofBorrelia burgdorferi DNA in tissues of infected mice after antibiotic treatment. J. Infect. Dis. 5 (1994) 1312–1316.

    Google Scholar 

  15. 15.

    DeKoning, J., Bosma, R. B., Hoogkamp-Korstanje, J. A. A. Demonstration of spirochetes in patients with Lyme disease with a modified silver stain. J. Med. Microbiol. 23 (1987) 261–267.

    Google Scholar 

  16. 16.

    Magnarelli, L. A., Meegan, J. M., Anderson, J. F., Chappeli, W. A. Comparison of indirect fluorescent antibody test with an enzyme-linked immunosorbent assay for serological studies of Lyme disease. J. Clin. Microbiol. 20 (1984) 181–184.

    Google Scholar 

  17. 17.

    Dorward, D. W., Schwan, T. G., Garon, C. F. Immune capture and detection ofBorrelia burgdorferi antigens in urine, blood, or tissues from infected ticks, mice, dogs and humans. J. Clin. Microbiol. 29 (1991) 1162–1170.

    Google Scholar 

  18. 18.

    Nielsen, S. L., Young, K. K. Y., Barbour, A. G. Detection ofBorrelia burgdorferi DNA by the polymerase chain reaction. Mol. Cell. Probes 4 (1990) 73–79.

    Google Scholar 

  19. 19.

    Guy, E. C., Stanek, G. Detection ofBorrelia burgdorferi in patients with Lyme disease by the polymerase chain reaction. J. Clin. Pathol. 44 (1991) 610–611.

    Google Scholar 

  20. 20.

    Rosa, P. A., Schwan, T. G. A specific and sensitive assay for the Lyme disease spirocheteBorrelia burgdorferi using the polymerase chain reaction. J. Infect. Dis. 160 (1989) 1018–1029.

    Google Scholar 

  21. 21.

    Huppertz, H. I., Schmidt, H., Karch, H.: Detection ofBorrelia burgdorferi by nested polymerase chain reaction in cerebrospinal fluid and urine of children with neuroborreliosis. Eur. J. Pediatr. (1993) 414–417.

  22. 22.

    Keller, T. L., Halperin, J. J., Whitman, M. PCR detection ofBorrelia burgdorferi DNA in cerebrospinal fluid of Lyme neuroborreliosis patients. Neurology 42 (1992) 32–42.

    Google Scholar 

  23. 23.

    Goodman, J. L., Jurkovich, P., Kramber, J. M., Johnson, R. C. Molecular detection of persistentBorrelia burgdorferi in the urine of patients with active Lyme disease. Infect. Immun. 59 (1991) 269–278.

    Google Scholar 

  24. 24.

    Liebling, M. R., Nishio, M. J., Rodriguez, A., Sigal, L. H., Jin, T., Louie, J. S. The polymerase chain reaction for the detection ofBorrelia burgdorferi in human body fluids. Arthritis Rheum. 36 (1993) 665–675.

    Google Scholar 

  25. 25.

    Lebech, A. M., Hansen, K. Detection ofBorrelia burgdorferi DNA in urine samples and cerebrospinal samples from patients with early and late Lyme neuroborreliosis by polymerase chain reaction. J. Clin. Microbiol. 30 (1992) 1646–1653.

    Google Scholar 

  26. 26.

    Walsh, P. S., Metzger, D. A., Higuchi, R. CHELEX 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material. Biotechniques 10 (1991) 506–513.

    Google Scholar 

  27. 27.

    Chiba, A., Takahashi, T., Nau, M. M., D'Amico, D., Curiel, D. T., Mitsudomi, T., Buchhagen, D. L., Carbone, D., Piantadosi, S., Doga, H., Reissman, P. T., Slamon, D. J., Holmes, E. C., Minna, J. D. Mutations in the p53 gene are frequent in primary, resected non-small cell lung cancer. Oncogene 5 (1990) 1603–1610.

    Google Scholar 

  28. 28.

    Sambrook, J., Fritsch, E. F., Maniatis, T. Molecular cloning. Cold Spring Harbor Laboratory Press, Cold Spring Harbor 1989, Vol. 3.

    Google Scholar 

  29. 29.

    Rys, P. N., Persing, D. H. Preventing false positives: quantitative evaluation of three protocols for inactivation of polymerase chain reaction amplification products. J. Clin. Microbiol. 31 (1993) 2356–2360.

    Google Scholar 

  30. 30.

    Barbour, A. G. Isolation and cultivation of Lyme disease spirochetes. Yale J. Biol. Med. 57 (1984) 521–525.

    Google Scholar 

  31. 31.

    Preac Mursic, V. Therapy ofLyme borreliosis. Antibiotic susceptibility ofBorrelia burgdorferi in vitro andin vivo. In:Weber, K., Burgdorfer, W. (eds.): Aspects ofLyme borreliosis. Springer-Verlag, Berlin 1993, pp. 301–311.

    Google Scholar 

  32. 32.

    Preac Mursic, V., Weber, K., Pfister, W., Wilske, B., Gross, B., Baumann, A., Prokop, J. Survival ofBorrelia burgdorferi in antibiotically treated patients with Lyme borreliosis. Infection 17 (1989) 355.

    Google Scholar 

  33. 33.

    Karch, H., Huppertz, H.-I., Böhme, M., Schmidt, H., Wiebecke, D., Schwarzkop, A. Demonstration ofBorrelia burgdorferi DNA in urine samples from healthy humans whose sera containB. burgdorferi-specific antibodies. J. Clin. Microbiol. 32 (1994) 2312–2314.

    Google Scholar 

  34. 34.

    Telford III, S. R., Dawson, J. E., Katavolos, P., Warner, C. K., Kolbert, C. P., Persing, D. H. Perpetuation of the agent of human granulocytic ehrlichiosis in a deer tick-rodent cycle. Proc. Natl. Acad. Sci. USA 93 (1996) 6209–6214.

    Google Scholar 

  35. 35.

    Magnarelli, L. A., Dumler, J. S., Anderson, J. F., Johnson, R. C., Fikrig, E. Coexistence of antibodies of tick-borne pathogenes of babesiosis, ehrilichiosis and Lyme borreliosis in human sera. J. Clin. Microbiol. 33 (1995) 3054–3057.

    Google Scholar 

  36. 36.

    Barbour, A. G., Maupin, G. O., Teltow, G. J., Carter, C. J., Piesman, J. Identification of an uncultivableBorrelia species in the hard tickAmblyomma americanum: Possible agent of a Lyme disease-like illness. J. Infect. Dis. 173 (1996) 403–409.

    Google Scholar 

  37. 37.

    Krause, P. J., Telford III, S. R., Spielman, A., Sikand, V., Ryan, R., Christianson, D., Burke, G., Brassard, P., Pollack, R., Peck, J., Persing, D. H. Concurrent Lyme disease and babesiosis. JAMA 275 (1996) 1657–1660.

    Google Scholar 

Download references

Author information

Affiliations

Authors

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Bayer, M.E., Zhang, L. & Bayer, M.H. Borrelia burgdorferi DNA in the urine of treated patients with chronic lyme disease symptoms. A PCR study of 97 cases. Infection 24, 347–353 (1996). https://doi.org/10.1007/BF01716077

Download citation

Keywords

  • Lyme Disease
  • Borrelia Burgdorferi
  • Outer Surface Protein
  • Chronic Lyme Disease
  • Lyme Disease Symptom