Background: Avian Influenza (AIV) and Newcastle Disease Viruses (NDV) are the two most important culprit in poultry diseases causing serious economic loss in the poultry industry in the world as well as in Bangladesh. However, there is no study for the simultaneously detection of AIV and NDV using rapid diagnostic kit in Bangladesh. Therefore, the present study was carried out to determine the prevalence of Avian Influenza and Newcastle Disease Viruses in poultry in the selected areas in Bangladesh. Methods: Avian Influenza (AIV) and Newcastle Disease Viruses (NDV) were detected using rapid antigen detection kit from field samples of poultry in Bangladesh. The cloacal swabs were collected from randomly selected 10 birds (broiler, layer, and native chicken, and duck) to detect AIV and NDV in four different districts or areas, and a total of 160 field samples were successfully tested in this study. Findings: Overall prevalence of AIV and NDV in different species of birds was 6.8% and 38.1%, respectively. The prevalence of AIV in layer and broiler bird was 15.0% and 12.5%, respectively. The prevalence of NDV in layer, broiler, native bird and duck was 37.5%, 32.5%, 55.0% and 27.5%, respectively. The prevalence of AIV in Dhaka, Gazipur, Mymensingh and Bogra districts were 12.5%, 12.5%, 0.0%, and 2.5%, respectively. The prevalence of NDV in Dhaka, Gazipur, Mymensingh and Bogra districts were 50.0%, 45.0%, 25.0%, and 32.5%, respectively. In galliformes, the prevalence of AIV and NDV was 9.16% and 41.6%, respectively. In anseriformes prevalence of AIV and NDV was 0.0% and 27.5%, respectively. Conclusion: This is the first time that we are using these types of kit in Bangladesh and it’s concluded that the test is very easy and rapid, less laborious, less time consuming and non expensive for the detection and differentiation of AIV and NDV in Bangladesh.