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

Abstract

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 RT-PCR 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.