The study was done on economically very important New castle Disease using cross sectional study from February to April 2022 in selected districts of South Western Shoa Zone in Ethiopia where outbreaks of a disease frequently occurred and sample size was determined using cluster random sampling. So, the objectives of the study were to know the status of sero-prevalence and its risk factors. Serological Indirect ELISA test was used to detect viral antibody and Multivariable Logistic Regression was used for analysis. The result showed that the overall chicken level sero-prevalence was 4.43% (17/384, 95% CI: 2.78 - 6.97) and flock-level seroprevalence 11.20% (14/125, 95% CI: 6.79 – 17.92). Multivariable analysis at chicken level indicated that the odds of Newcastle disease infection was significantly high in cross chicken breeds, in purchased and female chickens. In addition it was high at flock level in cross breed chicken flocks or mixed breed flocks (P<0.05) in compare to local breed flocks. From chicken level risk factors; breed (P=0.019), chicken source (P=0.008), sex (P=0.001) and flock level risk factors, flock mixture (P=0.000) were significantly associated with Newcastle disease infection. The seroprevalence of NCDV in the backyard chicken production system may be due to field exposure of chickens to the disease and revealed the significance of the additional study on the serotype and strains of NCDV that are circulating in the study sites to plan suitable control and prevention actions.