Aims: The aim of the study is to develop the optimised parameters of electrochemical DNA biosensors for the specific detection of the Infectious Bronchitis Virus (IBV) in chickens. The goal is to further create a highly sensitive and specific biosensor that can be used for on-site monitoring of IBV on poultry farms. Methodology and results: In this study, an electrochemical DNA biosensor was developed for detecting a specific sequence in the IBV genome. The process involved attaching a NH2-ssDNA probe to a gold electrode, followed by hybridization with the target DNA. Various parameters like buffer, pH, scan rate, incubation time, redox indicators and temperature were optimised using cyclic voltammetry. The probe DNA was designed to enhance hybridization efficiency, which was assessed by measuring current signals. The biosensor, under optimal conditions, demonstrated high sensitivity and specificity when tested with different sequences, including complementary, non-complementary and mismatched ones. Cross-reactivity studies against non-IBV viruses showed distinguishable current signals. These findings have implications for developing a portable on-site IBV monitoring device for use on farms. Conclusion, significance and impact of study The optimised parameters and specificity of the electrochemical DNA biosensor suggest its potential for the development of a portable device for on-site monitoring of IBV on poultry farms. This device could prove to be a valuable tool for the early detection of IBV, helping to prevent further spread of the disease. However, it's essential to conduct further research to ensure the practicality and accuracy of the biosensor in real-world farm settings.