The present work was carried out to evaluate different quantitation techniques when Newcastle disease virus was intended to use as an oncolytic agent. The R2B Mukteshwar strain of NDV was procured as lyophilized vaccine. Then application were carried out like haemagglutination test, tissue culture infective dose-50 (TCID50), plaque forming unit (PFU) calculation and real time PCR to enumerate the number of viruses. The HA titre was obtained as 1:128 across the dilutions. While TCID50 and PFU counts were obtained as 1×108.16/ ml and 4.2×107 PFU/ ml, respectively. Based upon comparison with standard NDV RNA, Real-time PCR also revealed the number of virus 108/ ml. HA was found consistent but indirect; contrastingly TCID50 suffered with subjectivity of interpretation. PFU counts were found within a range and lease possibility of interpretation error than TCID50. Though real time was found automated, highly specific and sensitive assay but handling of RNA and cost were limiting factors.