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Updated: Aug 14, 2026

Detecting the Lyme Disease Spirochete, Borrelia Burgdorferi, in Ticks Using Nested PCR
Published on: February 4, 2018
Epidemiology and diagnosis of Lyme borreliosis
1Max von Pettenkofer-Institute, University of Munich, National Reference Center for Borreliae, Pettenkofer-Strasse 9a, D-80336 Munich, Germany. Bettina.Wilske@mvp-bak.med.uni-muenchen.de
Abstract:
The multisystem disease Lyme borreliosis is the most frequent tick-transmitted disease in the northern hemisphere. In Europe Lyme borreliosis is most frequent in Central Europe and Scandinavia (up to 155 cases per 100,000 individuals) and is caused by the species, B. burgdorferi sensu stricto, B. afzelii and B. garinii. The recently detected genospecies A14S may also play a role in skin manifestations. Microbiological diagnosis in European patients must consider the heterogeneity of borreliae for development of diagnostic tools. According to guidelines of the USA and Germany, serological diagnosis should follow the principle of a two-step procedure (enzyme-linked immunosorbent assay (ELISA) as first step, if reactive; followed by immunoblot). The sensitivity and standardization of immunoblots has been considerably enhanced by use of recombinant antigens (p100, p58, p41i, VlsE, OspC, DbpA) including those expressed primarily in vivo (VlsE and DbpA) instead of whole cell lysates. VlsE is the most sensitive antigen for IgG antibody detection, OspC for IgM antibody detection. At present, detection rates for serum antibodies are 20%-50% in stage I, 70%-90% in stage II, and nearly 100% in stage III Lyme disease. Detection of the etiological agent by culture or polymerase chain reaction (PCR) should be confined to specific indications and specialized laboratories. Recommended specimens are skin biopsy specimens, cerebrospinal fluid (CSF) and synovial fluid. The best results are obtained from skin biopsies with culture or PCR (50%-70%) and synovial tissue or fluid (50%-70% with PCR). CSF yields positive results in only 10%-30% of patients except when the duration of symptoms is shorter than 2 weeks (50% sensitivity). Methods which are not recommended or adequately documented for diagnosis are antigen tests on body fluids, PCR of urine, and lymphocyte transformation tests.
Insights
Accurate Lyme borreliosis diagnosis in Europe requires considering diverse Borrelia species. Enhanced serological tests using recombinant antigens improve detection rates across all disease stages.
Area of Science:
- Medical Microbiology
- Infectious Diseases
- Tick-borne Illnesses
Background:
- Lyme borreliosis is a prevalent tick-transmitted disease in the Northern Hemisphere, particularly in Central Europe and Scandinavia.
- Causative agents include *B. burgdorferi sensu stricto*, *B. afzelii*, and *B. garinii*, with genospecies A14S potentially involved in skin manifestations.
- Diagnostic strategies must account for the genetic diversity of European Borrelia species.
Purpose of the Study:
- To review current diagnostic approaches for Lyme borreliosis in European patients.
- To highlight the importance of considering Borrelia species heterogeneity in diagnostic tool development.
- To evaluate the efficacy of serological and etiological agent detection methods.
Main Methods:
- Review of serological diagnostic guidelines (USA and Germany) recommending a two-step approach: ELISA followed by immunoblot.
- Utilization of recombinant antigens (e.g., VlsE, OspC) in immunoblots for enhanced sensitivity and standardization compared to whole cell lysates.
- Assessment of direct detection methods like culture and PCR on specific specimens (skin biopsy, CSF, synovial fluid).
Main Results:
- Recombinant antigens, especially VlsE for IgG and OspC for IgM, significantly improve antibody detection rates.
- Serological detection rates increase with disease stage: 20%-50% (Stage I), 70%-90% (Stage II), nearly 100% (Stage III).
- PCR and culture yield best results from skin biopsies and synovial fluid (50%-70%), with lower sensitivity in CSF (10%-30%).
Conclusions:
- Modern serological diagnostics leveraging recombinant antigens offer improved sensitivity for Lyme borreliosis.
- Direct detection methods (PCR, culture) are valuable for specific clinical indications and sample types.
- Certain diagnostic methods, including antigen tests on body fluids and PCR of urine, are not recommended.
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