Detection by hemi-nested reverse transcription polymerase chain reaction and genetic characterization of wild type strains of Canine distemper virus in suspected infected dogs

Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
Cristina Esmeralda Di FrancescoF Marsilio

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.

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Citations

Feb 18, 2014·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Rebecca P WilkesMelissa A Kennedy
May 30, 2019·PloS One·Vivaldo Gomes da CostaRicardo Henrique Krüger
Jan 17, 2021·Viruses·Tiziana TroguAna Moreno

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Datasets Mentioned

BETA
GQ328902.1
GQ328903.1
GQ328904.1
DQ494319

Methods Mentioned

BETA
electrophoresis
PCR

Software Mentioned

FASTA
Basic Local Alignment Search Tool ( BLAST
MEGA
ClustalW2

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