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

Molecular detection of Borrelia burgdorferi sensu lato – An analytical comparison of real-time PCR protocols from five different Scandinavian laboratories.
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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 AC, Matussek A, Lindgren PE, Henningsson AJ,


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 AC, Matussek A, Lindgren PE, Henningsson AJ, (click to view)

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 AC, Matussek A, Lindgren PE, Henningsson AJ,

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PloS one 2017 09 2212(9) e0185434 doi 10.1371/journal.pone.0185434
Abstract
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.

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