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RESEARCH PRODUCT
Molecular detection of Borrelia burgdorferi sensu lato – An analytical comparison of real-time PCR protocols from five different Scandinavian laboratories
Gabriele MargosRimtas DargisAnn Cathrine PeterssonVolker FingerleMaximilian FallerPer-eric LindgrenVivian KjellandVivian KjellandKatharina OrnsteinÅShild Kristine AndreassenHanne QuarstenAnna J. HenningssonPeter WilhelmssonAndreas MatussekMalin LagerSølvi NoraasRam Benny Dessausubject
0301 basic medicinePhysiologyDenmarklcsh:MedicineArtificial Gene Amplification and ExtensionPathology and Laboratory MedicinePolymerase Chain ReactionBiochemistryNervous SystemRNA Ribosomal 16SMedicine and Health Scienceslcsh:ScienceDNA extractionCerebrospinal FluidLyme DiseaseMultidisciplinarySpirochetesbiologyNorwayLyme borreliosisRelapsing FeverBacterial PathogensBody FluidsNucleic acidsReal-time polymerase chain reactionRibosomal RNAMedical MicrobiologyPathogensAnatomyWater MicrobiologyTransmitted diseaseResearch ArticleCell biologyCellular structures and organellesBorrelia Burgdorferi030106 microbiologyTickReal-Time Polymerase Chain ReactionResearch and Analysis MethodsSensitivity and SpecificityMicrobiologyMicrobiology in the medical areaMicrobiology03 medical and health sciencesExtraction techniquesSensuBorreliaMikrobiologi inom det medicinska områdetMedical historyBorrelia burgdorferiMolecular Biology TechniquesNon-coding RNAMicrobial PathogensMolecular BiologySwedenBacteriaBorrelialcsh:ROrganismsBiology and Life Sciencesbacterial infections and mycosesbiology.organism_classificationRNA extraction030104 developmental biologyRNAlcsh:QRibosomesdescription
Introduction Lyme borreliosis (LB) is the most common tick transmitted disease in Europe. The diagnosis of LB today is based on the patient A 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. Funding Agencies|Futurum Academy for Healthcare; Region Jonkoping County; Division of Medical Diagnostics; Region of Jonkoping County; Interreg IVA Program ScandTick [167226]; Interreg V program ScandTick Innovation [20200422, 2015-000167]; NSTAND [PN 13-28]
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2017-01-01 | PLOS ONE |