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

Bidragets oversatte titel: Molekylær detektion af Borrelia burgdorferi, sensu lato. Analytisk sammenligning af real-time PCR protokoller fra 5 Skandinaviske laboratorier

Malin Lager*, Maximilian Faller, Peter Wilhelmsson, Vivian Kjelland, Åshild Andreassen, Rimtas Dargis, Hanne Quarsten, Ram Dessau, Volker Fingerle, Gabriele Margos, Sølvi Noraas, Katharina Ornstein, Ann Cathrine Petersson, Andreas Matussek, Per-Eric Lindgren, Anna J. Henningsson

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Abstrakt

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.

OriginalsprogEngelsk
Artikelnummere0185434
TidsskriftPLOS ONE
Vol/bind12
Udgave nummer9
Antal sider17
ISSN1932-6203
DOI
StatusUdgivet - 1. sep. 2017

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Emneord

  • Borrelia burgdorferi
  • PCR
  • Diagnostic accuracy
  • laboratory diagnosis

Citationsformater

Lager, M., Faller, M., Wilhelmsson, P., Kjelland, V., Andreassen, Å., Dargis, R., Quarsten, H., Dessau, R., Fingerle, V., Margos, G., Noraas, S., Ornstein, K., Petersson, A. C., Matussek, A., Lindgren, P-E., & Henningsson, A. J. (2017). Molecular detection of Borrelia burgdorferi sensu lato: An analytical comparison of real-time PCR protocols from five different Scandinavian laboratories. PLOS ONE, 12(9), [e0185434]. https://doi.org/10.1371/journal.pone.0185434