Diagnostics and genotyping of Canine parvovirus type 2 (CPV-2) from disease cases in south-eastern Poland

Abstract

Canine parvovirus type 2 is one of the most common causes of death among puppies. Despite preventive vaccination, the disease continues to be diagnosed. The aim of the study was to provide a molecular characterization of CPV-2 isolates found in southeastern Poland. Genetic CPV-2 material was isolated from the blood (n=10) and feces (n=50) of infected dogs. The presence of CPV-2 was confirmed by amplification of sequences coding both VP1 and VP2 protein. The products of the PCR reaction with primers amplifying VP2 protein were sequenced and used for genotyping. Bioinformatics analysis of the sequenced PCR product was performed to determine the phylogenetic relationships with variants recorded in the public databases. Based on the analysis of polymorphism in the nucleotide sequence 7 nucleotide variants were detected and assigned into four amino acid groups. Representatives of three groups contained asparagine at amino acid position 426 of the VP2 protein, which is characteristic of CPV-2a. The variant from the fourth group belonged to type CPV-2b. CPV-2a is the dominant antigenic type of CPV-2 in Poland. The pathogen’s high degree of polymorphism is manifested not only by the presence of numerous variants within the type, but also by the presence of representatives of type CPV-2b. Further studies of the molecular epidemiology of CPV-2 are necessary to optimize the effectiveness of preventive measures.

Keywords

• canine parvovirus type 2
• molecular diagnostics
• phylogenetics
• vp2 protein