Molecular detection of Borrelia burgdorferi sensu lato - An analytical comparison of real-time PCR protocols from five different Scandinavian laboratories

PLoS One. 2017 Sep 22;12(9):e0185434. doi: 10.1371/journal.pone.0185434. eCollection 2017.


Introduction: Lyme borreliosis (LB) is the most common tick transmitted disease in Europe. The diagnosis of LB today is based on the patient´s medical history, clinical presentation and laboratory findings. The laboratory diagnostics are mainly based on antibody detection, but in certain conditions molecular detection by polymerase chain reaction (PCR) may serve as a complement.

Aim: The purpose of this study was to evaluate the analytical sensitivity, analytical specificity and concordance of eight different real-time PCR methods at five laboratories in Sweden, Norway and Denmark.

Method: Each participating laboratory was asked to analyse three different sets of samples (reference panels; all blinded) i) cDNA extracted and transcribed from water spiked with cultured Borrelia strains, ii) cerebrospinal fluid spiked with cultured Borrelia strains, and iii) DNA dilution series extracted from cultured Borrelia and relapsing fever strains. The results and the method descriptions of each laboratory were systematically evaluated.

Results and conclusions: The analytical sensitivities and the concordance between the eight protocols were in general high. The concordance was especially high between the protocols using 16S rRNA as the target gene, however, this concordance was mainly related to cDNA as the type of template. When comparing cDNA and DNA as the type of template the analytical sensitivity was in general higher for the protocols using DNA as template regardless of the use of target gene. The analytical specificity for all eight protocols was high. However, some protocols were not able to detect Borrelia spielmanii, Borrelia lusitaniae or Borrelia japonica.

Publication types

  • Comparative Study
  • Multicenter Study

MeSH terms

  • Borrelia burgdorferi / genetics*
  • Cerebrospinal Fluid / microbiology
  • Denmark
  • Lyme Disease / diagnosis
  • Norway
  • RNA, Ribosomal, 16S / genetics
  • Real-Time Polymerase Chain Reaction / methods*
  • Relapsing Fever / microbiology
  • Sensitivity and Specificity
  • Sweden
  • Water Microbiology


  • RNA, Ribosomal, 16S

Grant support

Part of this work has received financial support from Futurum Academy for Healthcare, Region Jönköping County, Division of Medical Diagnostics, Region of Jönköping County, Interreg IV A Program ScandTick (grant no. 167226), Interreg V program ScandTick Innovation t (project ID. 20200422, reference no. 2015-000167) and INSTAND (PN 13-28). Founders had no role in the study design, data collection, analysis and interpretation of data, writing and preparation of the report and in decision to submit the article for publication.