RT-PCR based assay for detection of Newcastle disease virus isolated from poultry in Kashmir
Rafia Maqbool, Shakil A Wani, Asifa Wali, Zahid Kashoo, Pervaiz Dar, Sabia Qureshi, Isfaqul Hussain, Aazima Shah, Parvaiz Sikander, MY Ganaie and Saba Bukhari
Newcastle disease (ND) caused by a virulent strain of Avian paramyxovirus-1(vAPMV-1), is included in List A of the Office International des Epizooties and is a highly contagious disease of poultry. ND remains a major problem in existing or developing poultry industries. Confirmatory diagnosis requires isolation and detection of the virus. The present study was carried out to detect the Newcastle disease virus by RT PCR based assay isolated from poultry in Kashmir. 9 out of 10 disease outbreaks suspected for ND were positive. The clinical and postmortem lesions were characteristic of ND. The virus was isolated on the chicken embryo fibroblast (CEF) cell line. The virus was able to replicate easily in the cell line without supplementing with any trypsin-like protease. The detection was carried out by amplification of 356 bp hypervariable region of Fusion protein gene with F0 cleavage site by RT-PCR which is regarded as a major determinant of pathogenicity.