title: A duplex real-time PCR assay based on TaqMan technology for simultaneous detection and differentiation of canine adenovirus types 1 and 2.
abstract: Canine adenoviruses are a major cause of disease in dogs, coyotes, red foxes and wolves, as well as in other carnivores and marine mammals. Canine adenovirus type 1 (CAdV-1) and canine adenovirus type 2 (CAdV-2) cause infectious canine hepatitis (ICH) and infectious tracheobronchitis (ITB), respectively. In this study, a duplex real-time PCR assay for simultaneous detection and characterisation of CAdV-1 and CAdV-2 was developed by using a single primer pair and virus-specific probes. The assay was validated testing standard DNAs produced on purpose and clinical samples of various matrices known to be positive for CAdV-1, CAdV-2 or both viruses. Precise calculation of DNA loads in samples containing a wide range of viral amounts was allowed by generating a standard curve for absolute quantification. The assay was proven to be highly specific, since no cross-reactions with the different CAdV type was observed, and sensitive, being able to detect less than 10 copies of CAdV-1/CAdV-2 DNA. The low intra-assay and interassay coefficient of variations demonstrated a high repeatability, thus confirming the potential use of this assay for quantitative detection of CAdV-1 and CAdV-2 for rapid diagnosis and epidemiological investigations.
authors.affiliation: Department of Veterinary Medicine, University of Bari, Strada per Casamassima Km 3, 70010 Valenzano, Bari, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Dowgier
authors.affiliation: Department of Veterinary Medicine, University of Bari, Strada per Casamassima Km 3, 70010 Valenzano, Bari, Italy.
authors.forename: Viviana
authors.initials: V
authors.lastname: Mari
authors.affiliation: Department of Veterinary Medicine, University of Bari, Strada per Casamassima Km 3, 70010 Valenzano, Bari, Italy.
authors.forename: Michele
authors.initials: M
authors.lastname: Losurdo
authors.affiliation: Istituto Zooprofilattico Sperimentale di Puglia e Basilicata, via Manfredonia 20, 71100 Foggia, Italy.
authors.forename: Vittorio
authors.initials: V
authors.lastname: Larocca
authors.affiliation: Istituto Zooprofilattico Sperimentale di Puglia e Basilicata, via Manfredonia 20, 71100 Foggia, Italy.
authors.forename: Maria Loredana
authors.initials: ML
authors.lastname: Colaianni
authors.affiliation: Department of Veterinary Medicine, University of Bari, Strada per Casamassima Km 3, 70010 Valenzano, Bari, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Cirone
authors.affiliation: Department of Veterinary Medicine, University of Bari, Strada per Casamassima Km 3, 70010 Valenzano, Bari, Italy.
authors.forename: Maria Stella
authors.initials: MS
authors.lastname: Lucente
authors.affiliation: Department of Veterinary Medicine, University of Bari, Strada per Casamassima Km 3, 70010 Valenzano, Bari, Italy.
authors.forename: Vito
authors.initials: V
authors.lastname: Martella
authors.affiliation: Department of Veterinary Medicine, University of Bari, Strada per Casamassima Km 3, 70010 Valenzano, Bari, Italy; Istituto Zooprofilattico Sperimentale di Puglia e Basilicata, via Manfredonia 20, 71100 Foggia, Italy.
authors.forename: Canio
authors.initials: C
authors.lastname: Buonavoglia
authors.affiliation: Department of Veterinary Medicine, University of Bari, Strada per Casamassima Km 3, 70010 Valenzano, Bari, Italy. Electronic address: nicola.decaro@uniba.it.
authors.forename: Nicola
authors.initials: N
authors.lastname: Decaro
mesh_terms: D006516:Adenoviruses, Canine / Q000276:immunology / Q000302:isolation & purification*; D000818:Animals; D017931:DNA Primers; D004285:Dogs; D006522:Hepatitis, Infectious Canine / Q000175:diagnosis* / Q000821:virology; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers
keywords: Canine adenovirus 1; Canine adenovirus 2; Dog; Duplex real-time PCR