Development and validation of an indirect ELISA based on recombinant N-P fusion protein for detecting antibodies against parrot bornavirus in psittacine birds.

New iELISA provides reliable serological detection for subclinical parrot bornavirus infections.

This study presents development and validation of an indirect ELISA (iELISA) for serological detection of parrot bornavirus (PaBV) antibodies in psittacine birds. Researchers created recombinant nucleoprotein (N), phosphoprotein (P), and N-P fusion proteins derived from PaBV-4, identifying immunodominant epitopes through in silico prediction. The N-P fusion protein (48kDa) demonstrated superior performance with highest signal intensity and sensitivity. The optimized iELISA achieved 92.9% sensitivity and 96.7% specificity using a sample-to-positive ratio cutoff of 0.2064, with ROC curve analysis yielding an AUC of 0.985, indicating excellent diagnostic accuracy. Notably, no cross-reactivity was observed with other avian pathogens. Testing 208 serum samples from captive psittacine birds in Thailand revealed 68.27% PaBV seroprevalence, suggesting widespread subclinical infection. This diagnostic tool addresses significant challenges in detecting asymptomatic PaBV infections and proventricular dilatation disease (PDD), providing veterinarians with a reliable, practical alternative to complement molecular diagnostic methods for improved surveillance and disease control in psittacine populations.

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