title: Immunohistochemistry and real-time PCR as diagnostic tools for detection of Borrelia burgdorferi sensu lato in ticks collected from humans.
abstract: The objective of this study was to evaluate different methods used for detection of Borrelia burgdorferi sensu lato (s.l.) in ticks: immunohistochemistry followed by focus floating microscopy (FFM) and real-time polymerase chain reaction (real-time PCR) targeting the ospA and hbb genes. Additionally, an optimized ospA real-time PCR assay was developed with an integrated internal amplification control (IAC) for the detection of inhibition in the PCR assay and was validated as an improved screening tool for B. burgdorferi. One hundred and thirty-six ticks collected from humans in a hospital from Cluj-Napoca, Romania, were investigated regarding genus, stage of development and sex, and then tested by all three assays. A poor quality of agreement was found between FFM and each of the two real-time PCR assays, as assessed by concordance analysis (Cohen's kappa), whereas the agreement between the two real-time PCR assays was moderate. The present study argues for a low sensitivity of FFM and underlines that discordant results of different assays used for detection of B. burgdorferi in ticks are frequent.
authors.affiliation: Department of Infectious Diseases, University of Medicine and Pharmacy "Iuliu Haţieganu", Street Iuliu Moldovan 23, Cluj-Napoca, Romania. briciu.tincuta@umfcluj.ro.|Department of Parasitology and Parasitic Diseases, Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine, Calea Mănăstur 3-5, Cluj-Napoca, Romania. briciu.tincuta@umfcluj.ro.|National Reference Center for Borrelia, Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, Oberschleißheim, Germany. briciu.tincuta@umfcluj.ro.
authors.forename: Violeta T
authors.initials: VT
authors.lastname: Briciu
authors.affiliation: Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, Oberschleißheim, Germany.
authors.forename: Daniela
authors.initials: D
authors.lastname: Sebah
authors.affiliation: Department of Pathology, Clinic of Infectious Diseases, Street Iuliu Moldovan 23, Cluj-Napoca, Romania.
authors.forename: Georgiana
authors.initials: G
authors.lastname: Coroiu
authors.affiliation: Department of Infectious Diseases, University of Medicine and Pharmacy "Iuliu Haţieganu", Street Iuliu Moldovan 23, Cluj-Napoca, Romania.
authors.forename: Mihaela
authors.initials: M
authors.lastname: Lupşe
authors.affiliation: Department of Infectious Diseases, University of Medicine and Pharmacy "Iuliu Haţieganu", Street Iuliu Moldovan 23, Cluj-Napoca, Romania.
authors.forename: Dumitru
authors.initials: D
authors.lastname: Cârstina
authors.affiliation: Department of Infectious Diseases, University of Medicine and Pharmacy "Iuliu Haţieganu", Street Iuliu Moldovan 23, Cluj-Napoca, Romania.
authors.forename: Doina F
authors.initials: DF
authors.lastname: Ţăţulescu
authors.affiliation: Department of Parasitology and Parasitic Diseases, Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine, Calea Mănăstur 3-5, Cluj-Napoca, Romania.
authors.forename: Andrei D
authors.initials: AD
authors.lastname: Mihalca
authors.affiliation: Department of Parasitology and Parasitic Diseases, Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine, Calea Mănăstur 3-5, Cluj-Napoca, Romania.
authors.forename: Călin M
authors.initials: CM
authors.lastname: Gherman
authors.affiliation: Department of Medical Informatics and Biostatistics, University of Medicine and Pharmacy "Iuliu Haţieganu", Street Pasteur 6, Cluj-Napoca, Romania.
authors.forename: Daniel
authors.initials: D
authors.lastname: Leucuţa
authors.affiliation: Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, Oberschleißheim, Germany.
authors.forename: Fabian
authors.initials: F
authors.lastname: Meyer
authors.affiliation: National Reference Center for Borrelia, Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, Oberschleißheim, Germany.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Hizo-Teufel
authors.affiliation: National Reference Center for Borrelia, Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, Oberschleißheim, Germany.
authors.forename: Volker
authors.initials: V
authors.lastname: Fingerle
authors.affiliation: Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, Oberschleißheim, Germany.
authors.forename: Ingrid
authors.initials: I
authors.lastname: Huber
mesh_terms: D000818:Animals; D015748:Borrelia burgdorferi Group / Q000235:genetics / Q000302:isolation & purification*; D003870:Dermacentor / Q000254:growth & development / Q000382:microbiology*; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D018884:Ixodes / Q000254:growth & development / Q000382:microbiology*; D026863:Ixodidae; D007814:Larva / Q000382:microbiology; D009758:Nymph / Q000382:microbiology; D060888:Real-Time Polymerase Chain Reaction; D012383:Romania
keywords: Borrelia burgdorferi; Immunohistochemistry; Internal amplification control; Real-time PCR; Romania; Ticks