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Detection of Mixed Infection of Avian Influenza and Newcastle Disease Viruses in Chickens in Indonesia by Immunopathologic Immunohistochemistry Double Staining
Raden Wasito*1, Hastari Wuryastuti2 and Bambang Sutrisno1
1Department of Pathology, Faculty of Veterinary Medicine, Gadjah Mada University, Yogyakarta, Indonesia; 2Department of Veterinary Internal Medicine, Faculty of Veterinary Medicine, Gadjah Mada University, Yogyakarta, Indonesia
*Corresponding author:


Avian influenza virus (AIV) and Newcastle disease virus (NDV) are systemic disease that is highly infectious with high morbidity and mortality rates, and highly detrimental to economic value in the domestic and commercial poultry industries. Generally, in the field there is a mixed infection of AIV and NDV, each of which is highly pathogenic. Clinical signs are difficult to distinguish, so immuno-pathological immunohistochemistry (IHC) double staining was developed with the ability to confirm the diagnosis of mixed infection. In the present study, 20 chickens infected naturally with clinical signs of torticollis and curled toe paralysis that had a mixture of AIV and NDV infections and there are variations of tropism distribution of AIV and NDV in tissues were used. The AIV and NDV were visualized by performing the double staining technique DAB and Fast-Red with monoclonal antibody anti-hemagglutinin AIV-mouse (AIVHA) and polyclonal antibody anti-hemagglutinin-neuraminidase NDV-rabbit (NDVHN) proteins. AIV infected-lungs or brains had red coloration (AIVHA) and NDV infected lungs or brains had brown coloration (NDVHN). The IHC double staining technique is sensitive and accurate in the confirmation of the diagnosis of a mixed infection to determine the difference between AIV and NDV.

To Cite This Article: Wasito R, Wuryastuti H and Sutrisno B, 2018. Detection of mixed infection of Avian Influenza and Newcastle Disease Viruses inchickens in Indonesia by immunopathologic immunohistochemistry doublé staining. Pak Vet J, 38(4): 442-445.  


ISSN 0253-8318 (Print)
ISSN 2074-7764 (Online)