Pathological and Molecular Characterization of Newcastle Disease Virus Isolated from Gallus gallus in Java, Indonesia

Abstract

Background: Known as the vital avian disease in Indonesia, Newcastle disease (ND) has surged all over Indonesia which cause an immense drop in poultry industry. This investigation intended to examine the pathological and molecular characteristics of the latest Newcastle disease virus (NDV) derived from Gallus gallus in Java, Indonesia.Methods: The samples were inoculated on the embryonic chicken eggs (ECEs), then identified using the hemagglutination test (HA test), insulated isothermal PCR (iiPCR) and hemagglutination inhibition test (HI test). It conducted the reverse transcription-polymerase chain reaction (RT-PCR) using MSF1 and MSF2 primers and followed by nucleotide sequencing for the fusion protein gene. B-cells epitope prediction analysis was applied using the Immune Epitope Database and Analysis Resource (IEDB) and VaxiJen v2.0 to predict its immunogenicity. In addition, it also performed the histopathological examination by applying hematoxylin and eosin staining.Result: The results identified two viruses, NDV-1 and NDV-2, based on the fusion protein gene. Significantly, the cleavage site motif of NDV1 is 112GRQGRL117 (avirulent) and whereas for NDV2 is 112RRRKRF117 (virulent). Furthermore, it also predicted the “CKMGSRPSTKNPAP” peptide from NDV1 could be an immunogenic epitope candidate with a BepiPred linear epitope prediction score of 17.08. In summary, it suggests that NDV1 isolate can be used as a reference for vaccine design in Indonesia.