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      Correlation of Culture Positivity, PCR Positivity, and Burden of Borrelia burgdorferi Sensu Lato in Skin Samples of Erythema Migrans Patients with Clinical Findings

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          Abstract

          Background

          Limited data are available regarding the relationship of Borrelia burden in skin of patients with erythema migrans (EM) and the disease course and post-treatment outcome.

          Methods

          We studied 121 adult patients with EM in whom skin biopsy specimens were cultured and analyzed by quantitative PCR for the presence of Borreliae. Evaluation of clinical and microbiological findings were conducted at the baseline visit, and 14 days, 2, 6, and 12 months after treatment with either amoxicillin or cefuroxime axetil.

          Results

          In 94/121 (77.7%) patients Borrelia was detected in skin samples by PCR testing and 65/118 (55.1%) patients had positive skin culture result (96.8% B. afzelii, 3.2% B. garinii). Borrelia culture and PCR results correlated significantly with the presence of central clearing and EM size, while Borrelia burden correlated significantly with central clearing, EM size, and presence of newly developed or worsened symptoms since EM onset, with no other known medical explanation (new or increased symptoms, NOIS). In addition, the logistic regression model for repeated measurements adjusted for time from inclusion, indicated higher Borrelia burden was a risk factor for incomplete response (defined as NOIS and/or persistence of EM beyond 14 days and/or occurrence of new objective signs of Lyme borreliosis). The estimated association between PCR positivity and unfavorable outcome was large but not statistically significant, while no corresponding relationship was observed for culture positivity.

          Conclusions

          Higher Borrelia burden in EM skin samples was associated with more frequent central clearing and larger EM lesions at presentation, and with a higher chance of incomplete response.

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          Most cited references18

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          Lyme disease.

          A Steere (2001)
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            The early clinical manifestations of Lyme disease.

            Lyme disease, caused by a tick-transmitted spirochete, typically begins with a unique skin lesion, erythema chronicum migrans. Of 314 patients with this skin lesion, almost half developed multiple annular secondary lesions; some patients had evanescent red blotches or circles, malar or urticarial rash, conjunctivitis, periorbital edema, or diffuse erythema. Skin manifestations were often accompanied by malaise and fatigue, headache, fever and chills, generalized achiness, and regional lymphadenopathy. In addition, patients sometimes had evidence of meningeal irritation, mild encephalopathy, migratory musculoskeletal pain, hepatitis, generalized lymphadenopathy and splenomegaly, sore throat, nonproductive cough, or testicular swelling. These signs and symptoms were typically intermittent and changing during a period of several weeks. The commonest nonspecific laboratory abnormalities were a high sedimentation rate, an elevated serum IgM level, or an increased aspartate transaminase level. Early Lyme disease can be diagnosed by its dermatologic manifestations, rapidly changing system involvement, and if necessary, by serologic testing.
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              Lyme borreliosis.

              Lyme borreliosis is the most common tick-transmitted disease in the northern hemisphere and is caused by spirochaetes of the Borrelia burgdorferi species complex. A complete presentation of the disease is an extremely unusual observation in which a skin lesion results from a tick bite and is followed by heart and nervous system involvement, and later on by arthritis. Late involvement of eye, nervous system, joints, and skin can also occur. The only sign that enables a reliable clinical diagnosis of Lyme borreliosis is erythema migrans. Other features of some diagnostic value are earlobe lymphocytoma, meningoradiculoneuritis (Garin-Bujadoux-Bannwarth syndrome), and acrodermatitis chronica atrophicans. The many other symptoms and signs have little diagnostic value. Microbial or serological confirmation of borrelial infection is needed for all manifestations of the disease except for typical early skin lesions. However, even erythema migrans might not be pathognomonic for Lyme borreliosis, especially in the southern part of the USA where there is no microbiological evidence for infection with the agent. Treatment with antibiotics is beneficial for all stages of Lyme borreliosis, but is most successful early in the course of the illness. Prevention relies mainly on avoiding exposure to tick bites but there is some interest in chemoprophylaxis and also in vaccine development following initial disappointments.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                9 September 2015
                2015
                : 10
                : 9
                : e0136600
                Affiliations
                [1 ]Department of Infectious Diseases, University Medical Center Ljubljana, Ljubljana, Slovenia
                [2 ]Institute for Biostatistics and Medical Informatics, Faculty of Medicine, Ljubljana, Slovenia
                [3 ]Vaccine R & D, Baxter Innovations, Orth an der Donau, A-2304, Austria
                University of Kentucky College of Medicine, UNITED STATES
                Author notes

                Competing Interests: The microbiological analyses were funded by Baxter, the former employer of I. Livey, M. O'Rourke and A. Traweger. M. O'Rourke and I. Livey hold Baxter stocks/shares and a patent for a vaccine against Lyme Borreliosis. F. Strle was a consultant to Baxter regarding Lyme vaccine (function ended in 2013) and a member of the steering committee of the ESCMID Study Group on Lyme Borreliosis/ESGBOR. Patent details are as follows: 1. I. Livey, M. A. O’Rourke, M. Schwendinger and B.A. Crowe (2011), chimeric OspA genes, proteins, and methods of use thereof 31315/44557A1. 2. I. Livey, M. A. O’Rourke, M. Schwendinger and B.A. Crowe (2011), chimeric OspA genes, proteins, and methods of use thereof 31315/44557A2. 3. P. N. Barrett, G. Aichinger, B.A. Crowe, I. Livey and N. Wressnig (2012), compositions comprising chimeric OspA molecules and methods of use thereof 31315/47123A. 4. 6745WO1 BX2011T00615 / 31315/44557A1 PCT PCT/US11/36525. 5. 6745U1US BX2011T00613 / 31315/44557A1 USSN 13/107,787. 6. 6745WO2 BX2011T00616 / 31315/44557A2 PCT PCT/US11/36533. 7. 6745U2US BX2011T00614 / 31315/44557A2 USSN 13/107,796. There are no further patents, products in development or marketed products to declare. This does not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials, as detailed online in the guide for authors.

                Conceived and designed the experiments: FS DS MO AT IL LL. Performed the experiments: MO AT IL DS VM PB DV. Analyzed the data: MO AT IL LL DS FS. Contributed reagents/materials/analysis tools: MO AT IL FS DS. Wrote the paper: DS FS LL MO.

                [¤a]

                Current address: CROMA-Pharma GmbH, Stockerauerstraße 181, A-2100, Leobendorf, Austria

                [¤b]

                Current address: Institute of Tendon and Bone Regeneration, SCI-TReCS, Paracelsus Medical University, Strubergasse 22, 5020, Salzburg, Austria

                Article
                PONE-D-15-23393
                10.1371/journal.pone.0136600
                4564201
                26352832
                bcb0fbd0-30e8-4ae9-a3d6-84ce6b999b06
                Copyright @ 2015

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

                History
                : 5 June 2015
                : 5 August 2015
                Page count
                Figures: 3, Tables: 2, Pages: 10
                Funding
                The microbiological analyses were funded by Baxter. The clinical part of the study was supported by the Slovenian Research Agency (grant number P3-0296). Baxter provided support in the form of salaries for authors MOR, AT and IL, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the 'author contributions' section.
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