title: Comparison of two PCR strategies for the detection of bovine papillomavirus.
abstract: Bovine papillomavirus (BPV) is a diverse group of double-stranded DNA oncogenic viruses, which have been detected in epithelial lesions and body fluids. Most studies of BPV infection rely on a single method for DNA detection; however the use of any single method or technique may underestimate the true prevalence of this virus. The purpose of this study was to compare two PCR strategies for the detection of BPV in skin lesions and fluids: these involve the use of BPV type-specific and consensus primers. Seventy-two cutaneous lesions, 57 blood samples and 59 semen samples were collected. PCR was used with the FAP consensus primers and BPV type-specific primers (for BPVs 2, 3, 4, 5, 8, 9 and 10), along with sequencing assays, to detect the BPV types. Phylogenetic analysis was carried out by means of the maximum likelihood method. It was found that both FAP and BPV type-specific primer sets could amplify BPV types of DNA in skin lesions, blood and semen samples. However, the BPV type-specific primers were more sensitive than the consensus primers and were able to detect co-infection of BPV in the samples. The consensus primers amplified five BPV types and were more suitable for detecting new putative BPV types. Thus, account should be taken of both PCR primer systems to identify co-infection, the presence of novel viruses, and avoid false-negative results.
authors.affiliation: Department of Genetics, Federal University of Pernambuco, 50740521 Pernambuco, Brazil. angel-jp@hotmail.com
authors.forename: M A R
authors.initials: MA
authors.lastname: Silva
authors.forename: M V A
authors.initials: MV
authors.lastname: Batista
authors.forename: N E
authors.initials: NE
authors.lastname: Pontes
authors.forename: E U D
authors.initials: EU
authors.lastname: Santos
authors.forename: L C A
authors.initials: LC
authors.lastname: Coutinho
authors.forename: R S
authors.initials: RS
authors.lastname: Castro
authors.forename: V Q
authors.initials: VQ
authors.lastname: Balbino
authors.forename: A C
authors.initials: AC
authors.lastname: Freitas
mesh_terms: D000818:Animals; D001769:Blood / Q000821:virology; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000821:virology*; D017931:DNA Primers / Q000235:genetics; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D027383:Papillomaviridae / Q000235:genetics / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012661:Semen / Q000821:virology; D012680:Sensitivity and Specificity; D012867:Skin / Q000821:virology; D012871:Skin Diseases / Q000175:diagnosis / Q000662:veterinary / Q000821:virology; D014773:Virology / Q000379:methods*
chemical_list: D017931:DNA Primers