title: Detection by hemi-nested reverse transcription polymerase chain reaction and genetic characterization of wild type strains of Canine distemper virus in suspected infected dogs.
abstract: A new highly sensitive and specific hemi-nested reverse transcription polymerase chain reaction (RT-PCR) assay was applied to detect nucleoprotein (NP) gene of Canine distemper virus (CDV) in samples collected from dogs showing respiratory, gastrointestinal, and neurological signs. Thirty-eight out of 86 samples were positive suggesting that despite the vaccination, canine distemper may still represent a high risk to the canine population. The 968 base pair (bp) fragments from the hemagglutinin (H) gene of 10 viral strains detected in positive samples were amplified and analyzed by restriction fragment length polymorphism (RFLP) using AluI and PsiI enzymes in order to differentiate among vaccine and wild-type CDV strains and to characterize the field viral strains. The products of the both enzymatic digestions allowed identification all viruses as wild strains of CDV. In addition, the RFLP analysis with AluI provided additional information about the identity level among the strains analyzed on the basis of the positions of the cleavage site in the nucleotide sequences of the H gene. The method could be a more useful and simpler method for molecular studies of CDV strains.
authors.affiliation: Department of Comparative Biomedical Sciences, Faculty of Veterinary Medicine of Teramo, P.zza A. Moro, 45-64100 Teramo, Italy. cedifrancesco@unite.it
authors.forename: Cristina E
authors.initials: CE
authors.lastname: Di Francesco
authors.forename: Daniela
authors.initials: D
authors.lastname: Di Francesco
authors.forename: Barbara
authors.initials: B
authors.lastname: Di Martino
authors.forename: Roberto
authors.initials: R
authors.lastname: Speranza
authors.forename: Domenico
authors.initials: D
authors.lastname: Santori
authors.forename: Andrea
authors.initials: A
authors.lastname: Boari
authors.forename: Fulvio
authors.initials: F
authors.lastname: Marsilio
mesh_terms: D000818:Animals; D001483:Base Sequence / Q000235:genetics; D004216:Distemper / Q000175:diagnosis* / Q000821:virology; D004217:Distemper Virus, Canine / Q000235:genetics*; D004285:Dogs; D008969:Molecular Sequence Data; D010802:Phylogeny; D012150:Polymorphism, Restriction Fragment Length / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity