Production of Newcastle Disease Polyclonal Antibody as the Alternative of Immunohistochemistry Primary Antibody against Newcastle Disease in Poultry

Newcastle disease (ND) is the most pathogenic viral infection in poultry. Furthermore, the availability of
laboratories that support the molecular diagnosis of ND is still limited in Indonesia. The present study aimed to
produce ND polyclonal antibody as the alternative of immunohistochemistry primary antibody against ND in
poultry. Two adult male New Zealand White rabbits weighed 2.5 kg were vaccinated seven days after the adaptation
using intraperitoneal injection of the ND live vaccine at multilevel doses weekly. The serum was collected
inactivated, and purified in the sixth week. A total number of 31 chicken samples were collected and their samples
of brain, lung, spleen, and intestine were tested using immunohistochemistry and Reverse Transcription Polymerase
Chain reaction (RT-PCR). The result showed that 19/31 (61%) were positive against immunohistochemistry and RTPCR and a total of 12/31 (39%) were negative. Based on the obtained results, immunohistochemistry using ND polyclonal antibody had a similar accuracy with RT-PCR. It can be concluded that ND polyclonal antibody produced by vaccination in the rabbit could be used as the alternative immunohistochemistry primary antibody for diagnosing ND in poultry.