The Phosphoprotein Gene from Canine Distemper Virus as Target in Viral Detection

The Canine Distemper Virus (CDV) is the causal agent of a severe multisystem disease of dogs and other carnivorous species. Canine Distemper (CD) is a highly contagious disease that can reach high morbidity and mortality. The wide spectrum of clinical signs makes the clinical diagnosis of the disease difficult and requires confirmation through laboratory tests. The molecular detection of CDV has advantages in the diagnosis of the disease. In the present work, the technique of Polymerase Chain Reaction-nested version-previous reverse transcription (n-RT-PCR) for the detection of the viral phosphoprotein gene (P gene) was implemented. As positive controls, RNA from vaccinal strains and samples positive to other CDV genes was used by RT-PCR. As negative controls, RNA was used from dog samples without clinical signs of the disease and as control of reactants, nuclease-free water. Subsequently, the technique was used in 10 field samples of dogs with clinical suspicion of the disease, detecting the viral genome in 100% of the samples analyzed, generating a band close to 430 bp, corresponding to the targeted target sequence of the phosphoprotein gene of the CDV. The amplification products were characterized by their great intensity and sharpness, not observing nonspecific amplification bands. Finally, the sequencing of the products obtained from the n-RT-PCR was performed, obtaining a high percentage of nucleotide identity with respect to the nucleotide sequences of the P gene stored in the GenBank®. This allowed establishing that the n-RT-PCR implemented in this Title Memory allows the detection of CDV. The implementation of this technique will allow the antemortem detection of the virus and, therefore, facilitate the diagnosis and early treatment of the disease.