Detection of Canine Parvovirus Type 2 by Designing Multiple Methods and Genetic Characterization in Iran

Area of Science:

• Veterinary Virology
• Molecular Diagnostics
• Canine Infectious Diseases

Background:

• Canine parvovirus type 2 (CPV-2) is a highly contagious pathogen primarily affecting young dogs.
• Accurate and rapid diagnostic methods are crucial for timely treatment and disease control.
• Existing diagnostic assays may be time-consuming or lack optimal sensitivity.

Purpose of the Study:

• To design and optimize an in-house Polymerase Chain Reaction (PCR) assay for rapid CPV-2 detection.
• To compare the performance of the in-house PCR assay against Real-Time PCR and Loop-mediated Isothermal Amplification (LAMP) assays.
• To conduct phylogenetic analysis of CPV-2 field isolates using the virulence gene vp2.

Main Methods:

• Development and optimization of an in-house PCR assay targeting the CPV-2 vp2 gene.
• Comparison of detection limits and specificity with Real-Time PCR and LAMP assays.
• Agarose gel electrophoresis for PCR product analysis and phylogenetic analysis of partial vp2 gene sequences.

Main Results:

• The in-house PCR assay demonstrated high specificity, with no amplification from negative control genomes.
• The limit of detection was comparable across all tested assays: 19 copies for in-house PCR and LAMP, and 21 copies for Real-Time PCR.
• Phylogenetic analysis of five field isolates revealed three distinct clusters, providing insights into viral diversity.

Conclusions:

• The developed in-house PCR assay is a specific, sensitive, and rapid method for detecting major CPV-2 pathogens.
• This assay can serve as a valuable supplement to existing diagnostic methods, potentially replacing slower laboratory techniques.
• Phylogenetic analysis is essential for understanding CPV-2 epidemiology and informing disease control strategies.