title: Interlaboratory comparison of real-time polymerase chain reaction methods to detect Coxiella burnetii, the causative agent of Q fever.
abstract: The bacterium Coxiella burnetii, which has a wide host range, causes Q fever. Infection with C. burnetii can cause abortions, stillbirth, and the delivery of weak offspring in ruminants. Coxiella burnetii infection is zoonotic, and in human beings it can cause chronic, potentially fatal disease. Real-time polymerase chain reaction (PCR) is increasingly being used to detect the organism and to aid in diagnosis both in human and animal cases. Many different real-time PCR methods, which target different genes, have been described. To assess the comparability of the C. burnetii real-time PCR assays in use in different European laboratories, a panel of nucleic acid extracts was dispatched to 7 separate testing centers. The testing centers included laboratories from both human and animal health agencies. Each laboratory tested the samples using their in-house real-time PCR methods. The results of this comparison show that the most common target gene for real-time PCR assays is the IS1111 repeat element that is present in multiple copies in the C. burnetii genome. Many laboratories also use additional real-time PCR tests that target single-copy genes. The results of the current study demonstrate that the assays in use in the different laboratories are comparable, with general agreement of results for the panel of samples.
authors.affiliation: Veterinary Laboratories Agency, Woodham Lane, Addlestone, Surrey KT15 3NB, United Kingdom. r.jones@vla.defra.gsi.gov.uk
authors.forename: Rebecca M
authors.initials: RM
authors.lastname: Jones
authors.forename: Stefan
authors.initials: S
authors.lastname: Hertwig
authors.forename: James
authors.initials: J
authors.lastname: Pitman
authors.forename: Richard
authors.initials: R
authors.lastname: Vipond
authors.forename: Anna
authors.initials: A
authors.lastname: Aspán
authors.forename: Göran
authors.initials: G
authors.lastname: Bölske
authors.forename: Conall
authors.initials: C
authors.lastname: McCaughey
authors.forename: James P
authors.initials: JP
authors.lastname: McKenna
authors.forename: Bart J
authors.initials: BJ
authors.lastname: van Rotterdam
authors.forename: Arnout
authors.initials: A
authors.lastname: de Bruin
authors.forename: Robin
authors.initials: R
authors.lastname: Ruuls
authors.forename: Rob
authors.initials: R
authors.lastname: Buijs
authors.forename: Hendrik-Jan
authors.initials: HJ
authors.lastname: Roest
authors.forename: Jason
authors.initials: J
authors.lastname: Sawyer
mesh_terms: D000818:Animals; D016997:Coxiella burnetii / Q000235:genetics / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards / Q000662:veterinary*; D011778:Q Fever / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics; D012418:Ruminants / Q000382:microbiology*; D015047:Zoonoses / Q000382:microbiology*
chemical_list: D004269:DNA, Bacterial