title: Development of a SYBR Green real-time PCR assay with melting curve analysis for simultaneous detection and differentiation of canine adenovirus type 1 and type 2.
abstract: Canine adenovirus type 1 (CAdV-1) and canine adenovirus type 2 (CAdV-2) cause infectious canine hepatitis (ICH) and infectious tracheobronchitis (ITB) in dogs, respectively. Cases of ICH have been documented in recent years and recent surveys have demonstrated a wide percentage of asymptomatic CAdV-1 infection in the canine population. Since both CAdV types are detectable in the same biological matrices, and viral coinfection with CAdV-1 and CAdV-2 are reported with high frequency, it is urgent to have available a rapid, highly sensitive and specific assay for the diagnosis of CAdV infection and distinction between CAdV-1 and CAdV-2. In order to detect canine adenovirus in biological samples and to rapidly distinguish the two viral types, a SYBR Green real-time PCR assay was optimized to discriminate CAdV-1 and CAdV-2 via a melting curve analysis. The developed assay showed high sensitivity and reproducibility and was highly efficient and specific in discriminating the two CAdV types. This reliable and rapid technique may represent a simple, useful and economic option for simultaneous CAdV types detection, which would be feasible and attractive for all diagnostic laboratories, both for clinical purposes and for epidemiological investigations.
authors.affiliation: Department of Veterinary Medical Sciences, Alma Mater Studiorum-University of Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia (BO), Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Balboni
authors.affiliation: Department of Veterinary Medical Sciences, Alma Mater Studiorum-University of Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia (BO), Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Dondi
authors.affiliation: Department of Veterinary Medical Sciences, Alma Mater Studiorum-University of Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia (BO), Italy.
authors.forename: Santino
authors.initials: S
authors.lastname: Prosperi
authors.affiliation: Department of Veterinary Medical Sciences, Alma Mater Studiorum-University of Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia (BO), Italy. Electronic address: mara.battilani@unibo.it.
authors.forename: Mara
authors.initials: M
authors.lastname: Battilani
mesh_terms: D000257:Adenoviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D006516:Adenoviruses, Canine / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D000818:Animals; D052160:Benzothiazoles; D003959:Diamines; D004283:Dog Diseases / Q000175:diagnosis* / Q000821:virology; D004285:Dogs; D025202:Molecular Diagnostic Techniques / Q000379:methods; D009930:Organic Chemicals / Q000378:metabolism; D011804:Quinolines; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013194:Staining and Labeling; D044366:Transition Temperature; D014730:Veterinary Medicine / Q000379:methods*
chemical_list: D052160:Benzothiazoles; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I
keywords: Canine adenovirus; Dog; Infectious canine hepatitis; Infectious tracheobronchitis; Melting curve analysis; Real-time PCR