The Rift Valley Fever Virus (RVFV) is a zoonotic disease that is spread by mosquitoes. The potential effects of RVFV on both public health and agriculture have led the CDC and the USDA to designate it as a Category "A" priority disease that overlaps with select agents. Vaccination plays a significant role in controlling RVF disease, as it does with other viral infections. Rapid and sensitive quality assurance procedures tailored to the vaccination strain are crucial for ensuring the vaccine performs as expected. In light of the difficulties caused by the present outbreaks, these protocols will prove useful. Using real-time PCR with SYBR Green and low ROX, this work attempted to produce a more accurate and efficient procedure for measuring the Rift Valley Fever virus vaccinal strain than the traditional tissue culture-based titration method. The amount of virus in the vaccinal strain and seven different samples of RVFV ZH-501 were measured using both the usual tissue culture method and real-time PCR. Afterwards, the outcomes were contrasted. The real-time PCR standard curve that was created displays the mean CT values, the linear equation (y = -3.4639x + 38.506), and the r-squared value (0.996). A correlation coefficient of 0.996 indicated that the amplification was 94.4 percent effective. Results from the real-time PCR were comparable to those from tissue culture titration, and there was no statistically significant difference (p > 0.05). In cases of viral quantification, the results showed that the real-time PCR assay is the way to go because it is quick, easy, accurate, simple, and cheap.