title: Technical and clinical validation of three commercial real-time PCR kits for the diagnosis of neuroborreliosis in cerebrospinal fluid on three different real-time PCR platforms.
abstract: This study reports the evaluation of the technical and clinical validation of the O-DiaBorburg kit (DIA), Borrelia burgdorferi PCR kit, ISEX (GENE), and Borrelia burgdorferi sensu lato Real-TM (SAC) for the diagnosis of neuroborreliosis in cerebrospinal fluid based on both Borrelia DNA and CSF samples from patients with clinical suspicion of neuroborreliosis. This validation study was done by analysing the kits on the Rotorgene Q (RGQ), CFX96, and LightCycler480 (LC480). For all kits, the linear range was larger on RGQ than on CFX96 and LC480. A good reproducibility was obtained for all assays on all instruments. Storage at -20 °C resulted in a decreased reproducibility for SAC. Results of the limit of detection (LOD95) experiments indicated a better sensitivity than described in the kit insert for all kits on all PCR platforms. No cross-reactivity was found for genetically related organisms nor for other pathogens which may be present in CSF. All species of the Borrelia burgdorferi sensu lato complex were detected with the GENE and SAC kits. The DIA kit failed to detect B. lusitaniae. The results seemed to indicate a better overall performance for the GENE kit on RGQ. However, its diagnostic value could not be confirmed in the clinical validation study, wherein none of the 103 CSF samples from clinical neuroborreliosis cases showed a positive real-time PCR result with the GENE kit analysed on RGQ.
authors.affiliation: Research group Lab4U, Group of Management and Technology, UC Leuven-Limburg, Agoralaan gebouw B bus 3, 3590, Diepenbeek, Belgium. liesbeth.maes@ucll.be.
authors.forename: L
authors.initials: L
authors.lastname: Maes
authors.affiliation: Research group Lab4U, Group of Management and Technology, UC Leuven-Limburg, Agoralaan gebouw B bus 3, 3590, Diepenbeek, Belgium.
authors.forename: T
authors.initials: T
authors.lastname: Carolus
authors.affiliation: Research group Lab4U, Group of Management and Technology, UC Leuven-Limburg, Agoralaan gebouw B bus 3, 3590, Diepenbeek, Belgium.|Translational Research Center for Gastrointestinal Disorders (TARGID), University Hospital Gasthuisberg, KULeuven, Leuven, Belgium.
authors.forename: V
authors.initials: V
authors.lastname: De Preter
authors.affiliation: Research group Lab4U, Group of Management and Technology, UC Leuven-Limburg, Agoralaan gebouw B bus 3, 3590, Diepenbeek, Belgium.
authors.forename: S
authors.initials: S
authors.lastname: Ignoul
authors.affiliation: Clinical Biology, Laboratory for Molecular Diagnostics, Campus Salvator, Jessa Hospital, Salvatorstraat 20, 3500, Hasselt, Belgium.
authors.forename: R
authors.initials: R
authors.lastname: Cartuyvels
authors.affiliation: Research group Lab4U, Faculty of Industrial Engineering, KU Leuven - Campus Diepenbeek, Agoralaan gebouw B, 3590, Diepenbeek, Belgium.
authors.forename: L
authors.initials: L
authors.lastname: Braeken
authors.affiliation: Research group Lab4U, Group of Management and Technology, UC Leuven-Limburg, Agoralaan gebouw B bus 3, 3590, Diepenbeek, Belgium.
authors.forename: P-J
authors.initials: PJ
authors.lastname: D'Huys
authors.affiliation: Department of Laboratory Medicine, University Hospitals Leuven, Leuven, Belgium.|Department of Microbiology and Immunology, KU Leuven, Leuven, Belgium.
authors.forename: V
authors.initials: V
authors.lastname: Saegeman
authors.affiliation: Clinical Microbiology, Laboratory of Microbiology, University Clinic St-Luc UCL, Avenue Mounier 10 F, 1200, Brussels, Belgium.
authors.forename: B
authors.initials: B
authors.lastname: Kabamba
authors.affiliation: Clinical Biology, Laboratory for Molecular Diagnostics, Campus Salvator, Jessa Hospital, Salvatorstraat 20, 3500, Hasselt, Belgium.
authors.forename: M
authors.initials: M
authors.lastname: Raymaekers
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015748:Borrelia burgdorferi Group / Q000235:genetics / Q000302:isolation & purification*; D002555:Cerebrospinal Fluid / Q000382:microbiology*; D002648:Child; D002675:Child, Preschool; D005260:Female; D006801:Humans; D020852:Lyme Neuroborreliosis / Q000175:diagnosis*; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D055815:Young Adult