Abstract: Pair of primers and a TaqMan probe were synthesized according to M gene conservative sequence of Newcastle disease virus. The positive recombinant plasmid containing M gene of NDV ZJ1 strain isolated from goose was used as a positive quantitative template to establish a standard curve. And then a rea1time fluorescent quantitative RTPCR assay was established. The method has a good linear relationship within the 106 to 101 copies, with which 3 copies·μL-1 of the virus nucleic acid can be detected in the initial template, and has similar sensitivity with traditional virus isolation methods. The conforming rate of positive sum and negative sum with traditional virus isolation method was 900% and 998% respectively in detecting 500 clinic cloacal swab samples. The result showed that the constructed method paved the way for the early and rapid detection of NDV as well as quantitative analysis for the infection degree of NDV.