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The Persistence of Spirochetal Nucleic Acids in Active Lyme Arthritis

John F. Bradley, BA; Russell C. Johnson, PhD; and Jesse L. Goodman, MD
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Requests for Reprints: Dr. Jesse Goodman, Section of Infectious Diseases, University of Minnesota School of Medicine, Box 250 UMHC, 420 Delaware Street SE, Minneapolis, MN 55455. Acknowledgments: The authors thank Allan Ross for expert technical assistance, Drs. Judith Wanschura, Mike Rethwell, David Strike, Elizabeth Arndt, Tom Stillman, David Rhude, Kathleen Wesa, Archie Skemp, Walter Dorman, David Zoshke, Ellen Shammash, and Douglas Hotvedt for referring either patients or synovial fluid samples; Drs. Nicole Lurie and Nancy Meryhew for reviewing the manuscript; Beth Wetak and Jodi Aasmundrud for preparing the manuscript; and Dave Mottet for preparing the figure.

Copyright ©2004 by the American College of Physicians

Ann Intern Med. 1994;120(6):487-489. doi:10.7326/0003-4819-120-6-199403150-00007
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Lyme disease, caused by the spirochete Borrelia burgdorferi, is a common tick-borne infection. Arthritis develops in approximately one half of untreated patients who have a history of erythema migrans [1] and occurs, albeit rarely, even after treatment [2]. Spirochetes have rarely been cultivated from synovial fluid [34] but have been noted near blood vessels in silver-stained sections from synovectomy specimens [5]. Because it is difficult to find spirochetes in affected joints, the host response [68] and specific class II immune response genes [9] have been suggested as major determinants in the pathogenesis of arthritis. The general failure to recover spirochetes, however, does not exclude a primary role for B. burgdorferi in initiating and maintaining the arthritic process. The polymerase chain reaction (PCR) is capable of detecting low numbers of organisms [1011]. Although we (unpublished data) and others [12] retrospectively detected B. burgdorferi DNA by PCR in stored synovial fluid, the extreme sensitivity of the PCR makes contamination of such samples and false-positive results a potential problem. Therefore, to test the hypothesis that noncultivatable B. burgdorferi persists in Lyme arthritis, we did controlled, prospective culture and PCR studies on synovial fluid from patients with Lyme arthritis.

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Figure 1.
Southern blot of polymerase chain reaction (PCR)-amplified samples from patients and controls.B. burgdorferitop leftbottom left

Synovial fluid samples 1, 5, 8, and 12 are from four patients with presumed Lyme arthritis. Sample 11 is a follow-up from the same patient who provided sample 12 but was obtained while the patient received antibiotic treatment. Positive control samples included 20 to 2000 organisms ( ); patient samples 1 to 4 were initially negative but then each spiked with 20 organisms ( ). Negative control samples included those from patients with other rheumatologic disorders (samples 2, 3, 4, 6, 7, 9, 10, and 13), human serum (sample 14), and laboratory controls in which known negative synovial fluid was extracted (X), or water, instead of sample DNA, was added in the laboratory (L) or to the final PCR master mix itself (N).

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