Development of an In-House Antigen ELISA Using Monoclonal and Polyclonal Antibodies Against an Isolated Canine Distemper Virus.

Abstract

Canine distemper virus (CDV) is a highly contagious pathogen that affects several animal families. While animals of all ages are susceptible, puppies face the highest risk. This study aimed to detect CDV in dogs with suspected infection, isolate the virus and develop a sensitive antigen-ELISA test using monoclonal and polyclonal antibodies. Samples were collected from 40 dogs and tested by RT-PCR, then cultured positive samples in MDCK cells for virus isolation. In addition, the CDV F protein gene sequence obtained from the MDCK-34388 isolate was added to GenBank. Phylogenetic analysis revealed that the isolate belongs to the Asia-1 genotype. Among the 40 tested dogs, 29 (72.5%) tested positive for CDV, exhibiting respiratory, neurological and mixed symptoms in 62.5%, 15% and 22.5% of cases, respectively. Using an inactivated CDV isolate, we produced monoclonal antibodies in BALB/c mice and polyclonal antibodies in New Zealand rabbits. For ELISA development, we HRP-labelled the 1G10-1A monoclonal antibody that targets the CDV F protein. The antigen ELISA test we developed showed 90% sensitivity and 87.5% specificity, with positive and negative predictive values of 87.8% and 89.74%, respectively. These findings indicate that the antigen ELISA test is a reliable diagnostic tool for CDV detection, complementing RT-PCR and supporting effective disease management for veterinary practice.